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    Metabolic Reconfiguration Activates Stemness and Immunomodulation of PDLSCs. International journal of molecular sciences Periodontal ligament derived stem cells (PDLSC) are adult multipotent mesenchymal-like stem cells (MSCs) that can induce a promising immunomodulation to interact with immune cells for disease treatment. Metabolic reconfiguration has been shown to be involved in the immunomodulatory activity of MSCs. However, the underlying mechanisms are largely unknown, and it remains a challenging to establish a therapeutic avenue to enhance immunomodulation of endogenous stem cells for disease management. In the present study, RNA-sequencing (RNA-seq) analysis explores that curcumin significantly promotes PDLSC function through activation of MSC-related markers and metabolic pathways. In vitro stem cell characterization further confirms that self-renewal and multipotent differentiation capabilities are largely elevated in curcumin treated PDLSCs. Mechanistically, RNA-seq reveals that curcumin activates ERK and mTOR cascades through upregulating growth factor pathways for metabolic reconfiguration toward glycolysis. Interestingly, PDLSCs immunomodulation is significantly increased after curcumin treatment through activation of prostaglandin E2-Indoleamine 2,3 dioxygenase (PGE2-IDO) signaling, whereas inhibition of glycolysis activity by 2-deoxyglucose (2-DG) largely blocked immunomodulatory capacity of PDLSCs. Taken together, this study provides a novel pharmacological approach to activate endogenous stem cells through metabolic reprogramming for immunomodulation and tissue regeneration. 10.3390/ijms23074038
    Curcumin promotes osteogenic differentiation of periodontal ligament stem cells through the PI3K/AKT/Nrf2 signaling pathway. Iranian journal of basic medical sciences OBJECTIVES:The aim of this study was to investigate the effect of curcumin on the osteogenic differentiation of human periodontal ligament stem cells (hPDLSCs) and its underlying potential mechanism. MATERIALS AND METHODS:The tissue explant adherence method was used to isolate hPDLSCs. Flow cytometry, Alizarin Red staining and Oil Red O staining were applied to confirm the stemness of the stem cells. CCK8 assays were used to evaluate the effect of curcumin at different concentrations on cytotoxicity, and alkaline phosphate (ALP) activity assays, ALP staining and Alizarin Red staining were used to measure the osteogenic differentiation ability. In addition, hPDLSCs were treated with LY294002 (a phosphatidylinositol-3-kinase [PI3K] inhibitor) and erythroid transcription factor NF-E2 siRNA (siNrf2), respectively in the presence of curcumin. Western blotting was applied to evaluate the protein kinase B (AKT) phosphorylation levels and the Nrf2 levels. Besides, western blotting, RT-qPCR, ALP activity assays, ALP staining and Alizarin Red staining were used to detect the potential effects of curcumin on osteogenic differentiation. RESULTS:Curcumin at an appropriate concentration had no cytotoxicity and could promote osteogenic differentiation of the hPDLSCs. The results of western blotting and RT-qPCR revealed that the protein and mRNA levels of ALP, COL1 and RUNX2 were increased by curcumin, while the PI3K/AKT/Nrf2 signaling pathway was activated. In addition, LY294002 was added to inhibit the PI3K/AKT signaling pathway, or siNrf2 was used to block the Nrf2 pathway; then, the stimulatory effects of curcumin on osteogenic differentiation were reversed. CONCLUSION:Curcumin could promote the osteogenesis of hPDLSCs, and the effect is related to the PI3K/AKT/Nrf2 signaling pathway. 10.22038/IJBMS.2020.44070.10351
    Curcumin promotes osteogenic differentiation of human periodontal ligament stem cells by inducting EGR1 expression. Shi Weiping,Ling Danhua,Zhang Feiyun,Fu Xiaohui,Lai Danping,Zhang Yanzhen Archives of oral biology OBJECTIVE:Human periodontal ligament stem cells (hPDLSCs) attract attention for the periodontal regeneration therapy. Curcumin may promote osteogenic differentiation of hPDLSCs. This research aims to elucidate whether Curcumin displays promoting osteogenic differentiation and its mechanism. METHODS:The hPDLSCs were isolated from human periodontal ligament by immunomagnetic beads, identified with immumofluorescence. hPDLSCs were treated with 0, 5, 10, 20, 50, 100 μmol/L Curcumin. The early growth response gene 1 (EGR1) siRNA or plasmind were tranfected into the hPDLSCs. The viability, Alkaline Phosphatase (ALP) activity and mineralizaiton level of hPDLSCs were measured with 3-(4,5)-dimethylthiahiazo(-z-y1)-3,5-di-phenytetrazoliumromide (MTT) assay, ALP Assay Kit or Alizarin Red staining. The expression of EGR1, RUNX family transcription factor 2 (Runx2), bone gamma-carboxyglutamate protein (OC), secreted phosphoprotein 1 (OPN) and collagen type I alpha 1 chain (Collagen I), in hPDLSC were determined by Western blotting and quantitative reverse transcription-polymerase chain reaction. RESULTS:The isolated hPDLSCs were spindle or irregular, arranged in radial shape and shown positive expression of STRO-1, CD146 and Vimentin. Curcumin 10 μmol/L treatment maximal promoting the cells viability, ALP activities, mineralization, and levels of Runx2, OC, OPN, Collagen I and EGR-1 in hPDLSCs. EGR-1 siRNA transfection inversed Curcumin's promoting effect on ALP activities, mineralization, and levels of Runx2, OC, OPN, Collagen I and EGR-1 in hPDLSCs. While the EGR-1 plasmid transfection enhanced Curcumin's promoting effect on these parameters of hPDLSCs. CONCLUSION:Curcumin promotes the osteogenic differentiation of hPDLSCs, which may work through the EGR1. Curcumin may be a promising medicine for periodontitis treatment and periodontal regeneration. 10.1016/j.archoralbio.2020.104958
    Synthesis, characterization, and evaluation of curcumin-loaded endodontic reparative material. Alipour Mahdieh,Fadakar Sadaf,Aghazadeh Marziyeh,Salehi Roya,Samadi Kafil Hossein,Roshangar Leila,Mousavi Ensieh,Aghazadeh Zahra Journal of biochemical and molecular toxicology Curcumin (CUR) is an ancient therapeutic agent with remarkable antimicrobial and anti-inflammatory properties. The purpose of the current study was to synthesize and evaluate a curcumin-based reparative endodontic material to reduce infection and inflammation besides the induction of mineralization during the healing of the dentin-pulp complex. Poly-ɛ-caprolactone (PCL)/gelatin (Gel)/CUR scaffold was synthesized and assessed by scanning electron microscopy, Fourier transform infrared spectroscopy, and thermo-gravimetric analysis (TGA). Agar diffusion test was performed against E. coli, A. baumannii, P. aeruginosa, S. aureus, E. faecalis, and S. mutans. Moreover, proliferative, antioxidative, anti-inflammatory, and calcification properties of these scaffolds on human dental pulp stem cells (hDPSCs) were evaluated. The results showed that PCL/Gel/CUR scaffold had antibacterial effects. Also, these CUR-based scaffolds had significant inhibitory effects on the expression of tumor necrosis factor α and DCF from inflamed hDPSCs (p < 0.05). Moreover, the induction of mineralization in hDPSCs significantly increased after seeding on CUR-based scaffolds (p < 0.05). Based on these findings, the investigated CUR-loaded material was fabricated successfully and provided an appropriate structure for the attachment and proliferation of hDPSCs. It was found that these scaffolds had antimicrobial, antioxidant, and anti-inflammatory characteristics and could induce mineralization in hDPSCs, which is essential for healing and repairing the injured dentin-pulp complex. 10.1002/jbt.22854
    Curcumin reduces apoptosis and promotes osteogenesis of human periodontal ligament stem cells under oxidative stress in vitro and in vivo. Tan Lingping,Cao Zeyuan,Chen Huan,Xie Yunyi,Yu Le,Fu Chuanqiang,Zhao Wei,Wang Yan Life sciences AIMS:Human periodontal ligament stem cells (hPDLSCs) tether the teeth to the surrounding bone and are considered as major functional stem cells responsible for regeneration of the alveolar bone and periodontal ligament tissue. However, the outcome of stem cell regenerative therapy is affected by the survival rate and their differentiation potential of transplanted cells. This is primarily because of local oxidative stress and chronic inflammation at the transplantation site. Therefore, our study aimed to explore whether a natural antioxidant, curcumin could increase the tissue regeneration ability of transplanted hPDLSCs. MAIN METHODS:A hydrogen peroxide environment and a rat cranial bone defect model were built to mimic the oxidative stress conditions in vitro and in vivo, respectively. We evaluated the effect of curcumin on oxidative status, apoptosis, mitochondrial function and osteogenic differentiation of HO-stimulated hPDLSCs in vitro. We also measured the effect of curcumin on cell viability and bone repair ability of transplanted hPDLSCs in vivo. KEY FINDINGS:Our data showed that curcumin enhanced cell proliferation, reduced the reactive oxygen species (ROS) levels and apoptosis, maintained the standard mitochondrial structure and function, and promoted osteogenic differentiation of HO-stimulated hPDLSCs. The extracellular regulated protein kinases 1/2 (Erk1/2) signaling pathway was determined to be involved in the osteogenic differentiation of the HO-stimulated hPDLSCs. Moreover, curcumin enhanced the viability and the bone repair ability of hPDLSCs in vivo. SIGNIFICANCE:Curcumin reduced apoptosis and promoted osteogenesis of the hPDLSCs under oxidative stress, and might therefore have a potential clinical use with respect to tissue regeneration. 10.1016/j.lfs.2021.119125
    3,4,5-Trihydroxybenzoic Acid Attenuates Ligature-Induced Periodontal Disease in Wistar Rats. Karatas Ozkan,Gevrek Fikret Anti-inflammatory & anti-allergy agents in medicinal chemistry BACKGROUND:3,4,5-Trihydroxybenzoic acid, which is also known as gallic acid, is an antiinflammatory agent that could provide beneficial effects in preventing periodontal inflammation. The present study aimed to evaluate the anti-inflammatory effects of gallic acid on experimental periodontitis in Wistar rats. Alveolar bone loss, osteoclastic activity, osteoblastic activity, and collagenase activity were also determined. METHODS:Thirty-two Wistar rats were used in the present study. Study groups were created as following: Healthy control (C,n=8) group; periodontitis (P,n=8) group; periodontitis and 30 mg/kg gallic acid administered group (G30,n=8); periodontitis and 60 mg/kg gallic acid administered group (G60,n=8). Experimental periodontitis was created by placing 4-0 silk sutures around the mandibular right first molar tooth. Morphological changes in alveolar bone were determined by stereomicroscopic evaluation. Mandibles were undergone histological evaluation. Matrix metalloproteinase (MMP)-8, tissue inhibitor of MMPs (TIMP)-1, bone morphogenetic protein (BMP)-2 expressions, tartrateresistant acid phosphatase (TRAP) positive osteoclast cells, osteoblast, and inflammatory cell counts were determined. RESULTS:The highest alveolar bone loss was observed in the periodontitis group. Both doses of gallic acid decreased alveolar bone loss as compared to the P group. TRAP-positive osteoclast cell counts were higher in the P group, and gallic acid successfully lowered these counts. Osteoblast cells also increased in gallic acid administered groups. Inflammation in the P group was also higher than those of C, G30, and G60 groups supporting the role of gallic acid in preventing inflammation. 30 and 60 mg/kg doses of gallic acid decreased MMP-8 levels and increased TIMP-1 levels. BMP levels increased in gallic acid administered groups, similar to several osteoblasts. CONCLUSION:Present results revealed an anti-inflammatory effect of gallic acid, which was indicated by decreased alveolar bone loss and collagenase activity and increased osteoblastic activity. 10.2174/1871523019666200206094335
    Caffeic acid phenethyl ester attenuates lipopolysaccharide-stimulated proinflammatory responses in human gingival fibroblasts via NF-κB and PI3K/Akt signaling pathway. Li Lei,Sun Wei,Wu Tao,Lu Rui,Shi Bin European journal of pharmacology Periodontal diseases often begin with chronic gingival inflammation, which causes the destruction of periodontal tissues. Inflammatory immune responses from host cells to bacteria, such as Porphyromonas gingivalis (P. gingivalis), cause periodontal degradation. Human gingival fibroblasts (HGFs) are the major cells in periodontal soft tissues. When stimulated by lipopolysaccharide (LPS), HGFs could secrete several pro-inflammatory cytokines and chemokines, such as interleukins (ILs) IL-6, IL-8, inducible nitric oxide synthase (iNOS), and cyclooxygenase 2 (COX-2). Caffeic acid phenethyl ester (CAPE) is the main active component of propolis, which is collected by honeybees from different plants and known for its anti-inflammatory effects. The anti-inflammatory effects of CAPE on the LPS-induced HGFs were demonstrated in this study. HGFs were pretreated with CAPE (10, 20, and 30µm) for 1h, followed by LPS stimulation (1μg/ml) for 24h. Enzyme-linked immunosorbent assay, Western blot analysis, and immunofluorescence staining were used to evaluate the production of IL-6, IL-8, iNOS, and COX-2, as well as the activation of TLR4-mediated NF-κB, PI3K/AKT, and MAPK signaling pathways. The results indicated that CAPE inhibits LPS-induced IL-6, IL-8, iNOS, and COX-2 production in a dose-dependent manner. Moreover, CAPE suppresses LPS-induced TLR4/MyD88 and nuclear factor kappa B (NF-κB) activation. In addition, phosphatidylinositol 3 kinase (PI3K) and protein kinase B (AKT) phosphorylation was inhibited by CAPE. These results demonstrated that CAPE could be effective for treating of periodontal diseases. 10.1016/j.ejphar.2016.11.003
    Is caffeic acid phenethyl ester more protective than doxycycline in experimental periodontitis? Yiğit Umut,Kırzıoğlu Fatma Yeşim,Uğuz Abdülhadi Cihangir,Nazıroğlu Mustafa,Özmen Özlem Archives of oral biology BACKGROUND AND OBJECTIVES:Host modulation therapies (anti-inflammatory drugs, bone-stimulating agents, anti-proteinase etc.) target the inhibition or stabilization of tissue breakdown. The aim of the present study was to evaluate the effects of caffeic acid phenethyl ester (CAPE) and/or low dose doxycycline (LDD) administrations on alveolar bone loss (ABL), serum cytokines and gingival apoptosis, as well as the levels of oxidants and anti-oxidants in rats with ligature-induced periodontitis. MATERIAL AND METHODS:The animals were randomly divided into five groups: Group C (periodontally healthy), Group PC (Periodontitis+CAPE), Group PD (Periodontitis+LDD), Group PCD (Periodontitis+CAPE+LDD), Group P (Periodontitis). Experimental periodontitis was induced for 14days. Levels of ABL, and the serum cytokines, interleukin (IL)-1 β, IL-6, tumor necrosis factor-α (TNF-α) and IL-10 were assessed as were the levels of the oxidants and anti-oxidants, malondialdehyde (MDA), glutathione (GSH) and glutathione peroxidase (GSH-Px), and levels of gingival apoptosis. RESULTS:The lowest ABL levels was evident in the PC group, among the experimental groups. There was also less inflammatory infiltration in the PC group than the PD group. IL-1β, IL-6, and IL-10 were lower in the PC group and higher in the P group in comparison to the levels in the other experiment groups. TNF-α levels in the PD group were higher than levels in the PC and PCD groups. The PC and PCD groups did not differ from the C group in regard to MDA levels. The highest GSH-Px level was found in the PC group. Gingival apoptosis in the PC group was not only lower than the PD and PCD groups, but also lower than in the C group. CONCLUSION:The present study suggests that CAPE has more anti-inflammatory, anti-oxidant and anti-apoptotic effects than LDD, with no additive benefits of a CAPE+LDD combination being evident in rats with periodontitis. 10.1016/j.archoralbio.2017.04.017
    A biochemical and immunohistochemical study of the effects of caffeic acid phenethyl ester on alveolar bone loss and oxidative stress in diabetic rats with experimental periodontitis. Kızıldağ Alper,Arabacı Taner,Albayrak Mevlüt,Balseven Havva Müge,Aksu Kızıldağ Canan,Tasdemir Ufuk Biotechnic & histochemistry : official publication of the Biological Stain Commission Caffeic acid phenethyl ester (CAPE) is used as a therapeutic agent to prevent bone loss. We determined the effects of systemically administered CAPE on alveolar bone loss and oxidative stress in diabetic rats with experimental periodontitis. Forty male rats were divided into four equal groups: control, experimental periodontitis (EP), EP-diabetes mellitus (EP-DM) and EP-DM-CAPE. DM was induced by streptozotocin, then lipopolysaccharide was injected to induce periodontitis. CAPE was administered to the EP-DM-CAPE group daily for 15 days. Then, serum samples were taken and the rats were sacrificed for histological analyses. Serum interleukin (IL-1β) and oxidative stress also were evaluated. Alveolar bone loss was assessed histomorphometrically. Alveolar bone loss and IL-1β levels were significantly less in the EP-DM-CAPE and EP groups compared to the EP-DM group. Oxidative stress was significantly less in the EP-DM-CAPE group compared to the EP and EP-DM groups. Receptor activator of nuclear factor kappa-B ligand (RANKL) levels were significantly higher in the EP-DM group compared to the disease groups. CAPE significantly reduced RANKL levels in the EP-DM-CAPE group compared to the EP-DM group. We found that CAPE treatment significantly inhibited DM induced oxidative stress and RANKL induced osteoclastogenesis and alveolar bone loss in diabetic rats with periodontitis. 10.1080/10520295.2020.1718756
    Protective effects of caffeic acid phenethyl ester on the heart in experimental periodontitis against oxidative stress in rats. Yiğit Umut,Kırzıoğlu Fatma Yeşim,Özmen Özlem,Uğuz Abdülhadi Cihangir Dental and medical problems BACKGROUND:Caffeic acid phenethyl ester (CAPE) may be considered as alternative treatment for periodontitis and benefit the heart by way of its ameliorative effects. OBJECTIVES:The aim of the study was to evaluate the effects of CAPE on cytokine levels and the oxidative status in the serum and heart tissue in a rat model of periodontitis. MATERIAL AND METHODS:Experimental animals were randomly assigned to 3 groups: control group (C; n = 8); periodontitis group (P; n = 10); and periodontitis + CAPE group (PC; n = 10). Caffeic acid phenethyl ester, at a dose of 10 μmol/kg/day, was administered by intraperitoneal injection over a 14-day period. Interleukin (IL)-1β, IL‑10 and tumor necrosis factor-alpha (TNF-α) were assessed in the serum. Glutathione (GSH), glutathione peroxidase (GSH-Px) and malondialdehyde (MDA) were assessed in both the serum and the heart tissue homogenate. RESULTS:Increased IL‑1β, IL‑10 and TNF-α serum levels were observed in the P group (p < 0.05). Caffeic acid phenethyl ester significantly decreased alveolar bone loss (ABL) and cytokine levels in the PC group (p < 0.05). Malondialdehyde, one of the strongest oxidants, was significantly decreased in the PC group as compared to the P group (p < 0.05). In both the serum and the heart tissue homogenate there were no differences in MDA levels between the PC and C groups. Furthermore, CAPE significantly increased GSH and GSH-Px levels in the serum and heart tissue (p < 0.05). CONCLUSIONS:Caffeic acid phenethyl ester has beneficial effects on the tissues affected by periodontitis. 10.17219/dmp/132388
    IL-8 as a Potential Therapeutic Target for Periodontitis and Its Inhibition by Caffeic Acid Phenethyl Ester In Vitro. Huang Yung-Kai,Tseng Kuo-Feng,Tsai Ping-Hsuan,Wang Jie-Sian,Lee Chang-Yu,Shen Ming-Yi International journal of molecular sciences Salivary levels of interleukin-8 (IL-8) are elevated in patients with periodontitis. Caffeic acid phenethyl ester (CAPE) improves the periodontal status in subjects. However, whether CAPE can reduce IL-8 expression is unclear. We collected saliva to determine proinflammatory cytokine levels and used subgingival calculus and surrounding tissues from patients with periodontitis for oral microbiota analysis via 16s ribosomal RNA gene sequencing. THP-1 cells were stimulated with sterile-filtered saliva from patients, and target gene/protein expression was assessed. IL-8 mRNA expression was analyzed in saliva-stimulated THP-1 cells treated with CAPE and the heme oxygenase-1 (HO-1) inhibitor tin-protoporphyrin (SnPP). In 72 symptomatic individuals, IL-8 was correlated with periodontal inflammation (bleeding on probing, = 0.45; < 0.001) and disease severity (bleeding on probing, = 0.45; < 0.001) but not with the four oral microbiota species tested. Reduced salivary IL-8 secretion was correlated with effective periodontitis treatment ( = 0.37, = 0.0013). In THP-1 cells, saliva treatment induced high IL-8 expression and IKK2 and nuclear factor-κB (NF-κB) phosphorylation. However, the IKK inhibitor BMS-345541, NF-κB inhibitor BAY 11-7082, and CAPE attenuated saliva-induced IL-8 expression. CAPE induced HO-1 expression and inhibited IKK2, IκBα, and NF-κB phosphorylation. Blocking HO-1 decreased the anti-inflammatory activity of CAPE. The targeted suppression of IL-8 production using CAPE reduces inflammation and periodontitis. 10.3390/ijms22073641
    Honokiol and Magnolol Inhibit CXCL10 and CXCL11 Production in IL-27-Stimulated Human Oral Epithelial Cells. Hosokawa Yoshitaka,Hosokawa Ikuko,Ozaki Kazumi,Matsuo Takashi Inflammation Honokiol and magnolol, which are lignans isolated from Magnolia quinquepeta, have some pharmacological effects. However, the anti-inflammatory effects of honokiol and magnolol on periodontal disease are still uncertain. The aim of this study was to examine the effect of honokiol and magnolol on CXC chemokine receptor 3 (CXCR3) ligands, which are related with Th1 cell migration, production in interleukin (IL)-27-stimulated human oral epithelial cells (TR146 cells). Honokiol and magnolol inhibited CXC chemokine ligand (CXCL)10 and CXCL11 production in IL-27-stimulated TR146 cells in a dose-dependent manner. Moreover, we revealed that honokiol and magnolol could suppress signal transducer and activator of transcription (STAT)3 and protein kinase B (Akt) phosphorylation in IL-27-stimulated TR146 cells though STAT1 phosphorylation was not suppressed by honokiol and magnolol treatment. Furthermore, STAT3 and Akt inhibitors could suppress CXCR3 ligand production in TR146 cells. In summary, honokiol and magnolol could reduce CXCR3 ligand production in oral epithelial cell by inhibiting STAT3 and Akt activation. 10.1007/s10753-018-0854-z
    Magnolol ameliorates the accumulation of reactive oxidative stress and inflammation in diabetic periodontitis. Liu Chia-Ming,Chen Szu-Han,Liao Yi-Wen,Yu Chuan-Hang,Yu Cheng-Chia,Hsieh Pei-Ling Journal of the Formosan Medical Association = Taiwan yi zhi BACKGROUND/PURPOSE:Periodontal disease and diabetes mellitus (DM) are both chronic inflammatory and highly prevalent diseases. A large amount of evidence suggested that the accumulation of oxidative stress plays a significant role in the deterioration of both diseases. Magnolol has been known to possess anti-inflammatory and anti-oxidant activities in various tissues, but its effects on gingival cells under diabetic conditions have not been fully understood. METHODS:We assessed the generation of reactive oxygen species (ROS), Transwell migration, and wound healing ability in response to the advanced glycation end products (AGEs) stimulation with or without Magnolol treatment. Subsequently, we examined the expression of Nrf2 and HO-1 to ascertain whether Magnolol was able to activate the anti-oxidant signaling. We also measured the secretion of IL-6 and IL-8, and conducted a knockdown experiment to elucidate the effect of Mrf2 on their secretion. RESULTS:The AGEs-induced ROS was dose-dependently downregulated following the Magnolol treatment. Likewise, the reduced Transwell migration and wound healing ability were improved by various concentrations of Magnolol. Results from qRT-PCR indicated that the suppression of Nrf2 and HO-1 following AGEs stimulation was reversed by Magnolol. Also, the AGEs-elicited production of IL-6 and IL-8 was inhibited by Magnolol. Moreover, our results demonstrated that this anti-inflammatory effect was mediated by the upregulation of Nrf2. CONCLUSION:These findings showed that excessive AGEs in the gingiva may lead to the accumulation of ROS and pro-inflammatory cytokines. Supplement of Magnolol may be beneficial to improve the impaired wound healing and inflammation by upregulation of Nrf2 signaling for DM patients with periodontal disease. 10.1016/j.jfma.2021.01.010
    Pterostilbene attenuates the proliferation and differentiation of TNF-α-treated human periodontal ligament stem cells. Yi Min,Wang Guanglei,Niu Jianhua,Peng Minghui,Liu Yi Experimental and therapeutic medicine Periodontitis is a common chronic inflammatory oral disease. The objective of periodontal treatment is to control infection whilst regenerating damaged periodontal tissue. The present study aimed to determine the potential effects of pterostilbene (PTE), a representative stilbene compound, on the proliferation and differentiation of human periodontal ligament stem cells (hPDLSCs). Different concentrations (1, 5, 10 and 20 µM) of PTE were applied to hPDLSCs, after which Cell Counting Kit-8 and western blotting assays were performed to examine the protein levels of Ki67, PCNA, p-IκBα, IκBα, Bcl-2, Bax, cleaved caspase3. The effect of PTE on the release of inflammatory factors, including IL-1β, IL-6 and IL-10 was assessed by RT-qPCR. The apoptosis of TNF-α-induced hPDLSCs was evaluated by TUNEL assay and western blotting. Additionally, the role of PTE in hPDLSC mineralization was evaluated using alizarin red staining. The expression levels of mineralization indices, including RUNX2 and ALP were subsequently determined using western blotting. Subsequently, the target of PTE was predicted using TargetNet database and AutoDock v4.2 software and verified using western blotting. The results of the present study revealed that PTE promoted the proliferation of hPDLSCs in a concentration-dependent manner. Furthermore, PTE treatment decreased the release of inflammatory factors and alleviated the apoptosis of TNF-α-induced hPDLSCs. PTE was also demonstrated to promote the formation of mineral nodules in TNF-α-induced hPDLSCs. The Targetnet database, along with molecular docking, indicated that histone deacetylases (HDACs) were the probable targets of PTE upstream of regulating periodontitis. The results of western blotting implied that TNF-α significantly increased expression levels of HDAC2, 4, 6 and 8, whilst PTE treatment markedly decreased HDAC4, 6 and 8 expression in a concentration-dependent manner compared with the TNF-α group, which further confirmed these conclusions. In summary, results of the present study revealed that PTE promoted TNF-α-induced hPDLSC proliferation and differentiation, whilst alleviating inflammation and apoptosis. PTE also inhibited the expression of HDACs, which may be involved in the mechanism of periodontitis. 10.3892/etm.2022.11233
    PINK1-mediated mitophagy reduced inflammatory responses to Porphyromonas gingivalis in macrophages. Oral diseases OBJECTIVE:Mitochondria are strained by microbial stimuli in the periodontal niche. Damaged mitochondria are cleared by mitophagy. The purpose of the study was to explore whether mitophagy participated in the progress of periodontitis and whether activation of mitophagy can inhibit inflammatory responses to bacterial infection in macrophages. METHODS:Mitophagy-related genes were measured in the healthy and inflamed human gingiva. Bone marrow-derived macrophages (BMDMs) were infected with Porphyromonas gingivalis. Dexmedetomidine, urolithin A, and resveratrol were used to activate mitophagy, while small interference RNA was utilized to knock down PTEN-induced putative protein kinase 1 (PINK1). Activation of mitophagy-related genes and colocalization of them were detected by Western blot and confocal imaging. Damages of mitochondria, accumulation of mitochondrial reactive oxygen species (mtROS), and production of IL-1β, IL-6, and TNF-α were measured. RESULTS:Levels of mitophagy-related genes were decreased in inflamed periodontal tissues and P. gingivalis-infected BMDMs. Dexmedetomidine, urolithin A, and resveratrol activated mitophagy, leading to reduced mitochondria damages, decreased mtROS generation, and inhibited IL-1β, IL-6, and TNF-α production. PINK1 knockdown reduced dexmedetomidine, urolithin A, and resveratrol-induced anti-inflammatory effect. CONCLUSION:Inhibited mitophagy participated in the progress of periodontitis. Activation of mitophagy may become a therapeutic target during the progress of periodontitis by reducing mtROS. 10.1111/odi.14286
    Chemoprevention of Experimental Periodontitis in Diabetic Rats with Silk Fibroin Nanoparticles Loaded with Resveratrol. Giménez-Siurana Ana,Gómez García Francisco,Pagan Bernabeu Ana,Lozano-Pérez Antonio Abel,Aznar-Cervantes Salvador D,Cenis José Luis,López-Jornet Pía Antioxidants (Basel, Switzerland) OBJECTIVE:the objective of the present work is to study the effectiveness of treatment with silk fibroin nanoparticles loaded with resveratrol in experimental periodontitis in a diabetic rat model. INTRODUCTION:Periodontitis is an inflammatory pathology highly related to other diseases, such as type II diabetes. Both diseases have a specific inflammatory condition, with Interleukin (IL)-6, IL-1β and Transforming Grow Factor (TGF)-1β being the most relevant proinflammatory factors. Silk fibroin (SF) nanoparticles loaded with resveratrol (Res-SFN) are a new alternative as a treatment. METHODS:40 diabetic Sprague Dawley male rats were used and periodontitis was induced by ligation. The animals were divided into 5 treatment groups, and 1 mL of treatment was administered once a day for 4 weeks. The groups were: I: Carboxymethyl cellulose (CMC) 0.8%, II: CMC 0.8% + SF 1%, III: CMC 0.8% + RES-SFN 3 mg/mL, IV: CMC 0.8% + SF 1% + RES-SFN 3 mg/mL, V: Water. A peripheral blood sample was taken every week to quantify the inflammatory profile by ELISA (IL-6, IL-1β and TGF-1β). After 4 weeks the sacrifice was carried out and biopsies of the gum were taken. RESULTS:Treatment with SF and RES-SFN reduced the amount of chemical inflammation mediators (with the exception of IL-1β in comparisons I-IV and II-IV ( > 0.05)), as well as the anatomopathological variables linked to it, in a significant way ( < 0.05). CONCLUSION:treatment with RES-SFN has reduced local inflammation in this experimental periodontitis model. 10.3390/antiox9010085
    In vitro evaluation of electrospun PLGA/PLLA/PDLLA blend fibers loaded with naringin for guided bone regeneration. Guo Zhenzhao,Wu Shuai,Li Hong,Li Qiyan,Wu Gang,Zhou Changren Dental materials journal The present study was to evaluate fiber mesh loaded with naringin via electrospinning to guide bone regeneration in vitro. The naringin-loaded fiber mesh was prepared via elctrospinning of PLGA, PLLA, PDLLA blending solution with naringin. SEM showed that naringin decreased the fiber's diameter according to the concentration of naringin. After 20 days' degradation in PBS, the drug-loaded fiber meshes still kept their stability with about 10% decrease in tensile strength. In vitro release experiments showed a sustained and steady naringin releasing profile with little initial burst releasing. Compared to the mats without naringin, the fiber mats loaded with naringin showed the most pronounced enhancement of cell growth when MC3T3-E1 cells were cultured on the fiber mats. The blend fiber loaded with naringin has optimized physical properties and sustained release profile in vitro. The study presents a promising fibrous mesh material for guided bone regeneration therapy. 10.4012/dmj.2016-220
    Black tea theaflavins attenuate Porphyromonas gingivalis virulence properties, modulate gingival keratinocyte tight junction integrity and exert anti-inflammatory activity. Ben Lagha A,Grenier D Journal of periodontal research BACKGROUND AND OBJECTIVE:Over the last 10 years, bioactive plant food compounds have received considerable attention in regard to their beneficial effects against periodontal disease. In this study, we investigated the effects of black tea theaflavins (TFs) on the virulence properties of Porphyromonas gingivalis and gingival keratinocyte tight junction integrity. In addition, the effects of black tea TFs on the nuclear factor-κB (NF-κB) signaling pathway and proinflammatory cytokine/matrix metalloproteinase (MMP) secretion by monocytes/macrophages were assessed. MATERIAL AND METHODS:Virulence factor gene expression in P. gingivalis was investigated by quantitative real-time PCR. A fluorescence assay was used to determine P. gingivalis adherence to, and invasion of, a gingival keratinocyte monolayer. Tight junction integrity of gingival keratinocytes was assessed by determination of transepithelial electrical resistance. Proinflammatory cytokine and MMP secretion by P. gingivalis-stimulated macrophages was quantified by ELISA. The U937-3xκB-LUC monocyte cell line transfected with a luciferase reporter gene was used to monitor NF-κB activation. Gelatin degradation was monitored using a fluorogenic assay. RESULTS:Black tea TFs dose-dependently inhibited the expression of genes encoding the major virulence factors of P. gingivalis and attenuated its adherence to gingival keratinocytes. A treatment of gingival keratinocytes with black tea TFs significantly enhanced tight junction integrity and prevented P. gingivalis-mediated tight junction damage as well as bacterial invasion. Black tea TFs reduced the secretion of interleukin (IL)-1β, tumor necrosis factor-α, IL-6, chemokine (C-X-C) ligand 8, MMP-3, MMP-8 and MMP-9 by P. gingivalis-stimulated macrophages and attenuated the P. gingivalis-mediated activation of the NF-κB signaling pathway. Lastly, black tea TFs inhibited gelatin degradation by MMP-9. CONCLUSION:This study provides clear evidence that black tea TFs represent promising multifunctional therapeutic agents for prevention and treatment of periodontal disease. 10.1111/jre.12411
    Quercetin Inhibits Inflammatory Response Induced by LPS in Human Gingival Fibroblasts via Suppressing NF-B Signaling Pathway. Xiong Gang,Ji Wansheng,Wang Fei,Zhang Fengxiang,Xue Peng,Cheng Min,Sun Yanshun,Wang Xia,Zhang Tianliang BioMed research international Quercetin, a natural flavonol existing in many food resources, has been reported to be an effective antimicrobial and anti-inflammatory agent for restricting the inflammation in periodontitis. In this study, we aimed to investigate the anti-inflammatory effects of quercetin on () lipopolysaccharide- (LPS-) stimulated human gingival fibroblasts (HGFs). HGFs were pretreated with quercetin prior to LPS stimulation. Cell viability was evaluated by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) assay. The levels of inflammatory cytokines, including interleukin-1 (IL-1), interleukin-6 (IL-6), and tumor necrosis factor- (TNF-), along with chemokine interleukin-8 (IL-8), were determined by enzyme-linked immunosorbent assay (ELISA). The mRNA levels of IL-1, IL-6, IL-8, TNF-, IB, p65 subunit of nuclear factor-kappa B (NF-B), peroxisome proliferator-activated receptor- (PPAR-), liver X receptor (LXR), and Toll-like receptor 4 (TLR4) were measured by real-time quantitative PCR (RT-qPCR). The protein levels of IB, p-IB, p65, p-p65, PPAR-, LXR, and TLR4 were characterized by Western blotting. Our results demonstrated that quercetin inhibited the LPS-induced production of IL-1, IL-6, IL-8, and TNF- in a dose-dependent manner. It also suppressed LPS-induced NF-B activation mediated by TLR4. Moreover, the anti-inflammatory effects of quercetin were reversed by the PPAR- antagonist of GW9662. In conclusion, these results suggested that quercetin attenuated the production of IL-1, IL-6, IL-8, and TNF- in LPS-treated HGFs by activating PPAR- which subsequently suppressed the activation of NF-B. 10.1155/2019/6282635
    ERK1/2 signaling mediated naringin-induced osteogenic differentiation of immortalized human periodontal ligament stem cells. Wei Kai,Xie Yuansheng,Chen Tianyu,Fu Bo,Cui Shaoyuan,Wang Yan,Cai Guangyan,Chen Xiangmei Biochemical and biophysical research communications Periodontal ligament stem cells (PDLSCs) are promising tools for the investigations of cell differentiation and bone regeneration. However, the limited life span significantly restricts their usefulness. In this study, we established an immortalized PDLSC cell line by the introduction of Bmi1 (PDLSC-Bmi1). Several genes related to cell cycle, cell replication and stemness were found to be changed with the overexpression of Bmi1. Compared with primary PDLSCs, the immortalized cells had a slower aging rate, maintained in a proliferative state without crisis for more than 30 passages, and retained the molecular markers and biological functions of primary ones. Using the PDLSC-Bmi1, we confirmed the promotive effect of naringin on osteogenesis. Naringin promoted the osteogenic differentiation of PDLSC-Bmi1 manifested as the increased activity of alkaline phosphatase (ALP), expression of the runt-related transcription factor 2 (Runx2) and osteocalcin (OCN), and formation of mineralized nodules. In addition, the extracellular regulated protein kinases (ERK) 1/2 was found to be activated by naringin, and the ERK1/2 specific inhibitor significantly inhibited naringin-induced osteogenic differentiation in PDLSC-Bmi1. Our results indicated that the overexpression of Bmi1 extended the life span of PDLSCs without perturbing their biological functions, and that naringin promoted the osteogenesis of PDLSC-Bmi1 at least partially through the ERK1/2 signaling pathway. 10.1016/j.bbrc.2017.05.130
    Effects of baicalin on the proliferation and expression of OPG and RANKL in human cementoblast-lineage cells. Journal of dental sciences BACKGROUND/PURPOSE:Baicalin, a natural bioactive flavonoid extracted from Georgi, mediates bone metabolism, and recent studies have revealed that it has cell signaling properties. However, its biological functions in cementoblasts still remain unclear. This study therefore aimed to investigate the effects of baicalin on bone resorption markers, including osteoprotegerin (OPG) and receptor activator of nuclear factor-κβ ligand (RANKL), in human cementoblast-lineage cells, as well as their proliferation ability. MATERIALS AND METHODS:Human cementoblast cell line (HCEM) cells were cultured and treated with 0, 0.01, 0.1, or 1 μM of baicalin. The proliferative capacity of cultured HCEM cells was analyzed using bromodeoxyuridine immunoassay and cell counting. The baicalin effect on OPG and RANKL expression was determined using quantitative polymerase chain reaction (qPCR) and western blotting. Furthermore, OPG expression was measured in 1 μM baicalin-treated HCEM cells in the presence or absence of the Wnt signaling pathway inhibitor, Dickkopf (Dkk)-1, using qPCR and western blotting. RESULTS:The addition of 0.01, 0.1, and 1 μM of baicalin did not significantly change the proliferative capacity of cultured HCEM cells. Compared with the non-supplemented group, baicalin increased and suppressed OPG and RANKL gene and protein expression, respectively, in a concentration-dependent manner. OPG mRNA and protein expression levels were increased by 1 μM baicalin, which was suppressed by Dkk-1 addition. CONCLUSION:Baicalin enhanced OPG expression in HCEM cells through the Wnt/beta-catenin signaling pathway, which could contribute to periodontal tissue regeneration. 10.1016/j.jds.2021.05.009
    Enhancing effects of myricetin on the osteogenic differentiation of human periodontal ligament stem cells via BMP-2/Smad and ERK/JNK/p38 mitogen-activated protein kinase signaling pathway. Kim Hyang-Yu,Park Sun-Young,Choung Se-Young European journal of pharmacology Myricetin is a flavonoid that found in berries, onions, and red grapes. It has been reported to have various pharmacological effects such as anti-inflammation, anti-oxidant and anti-cancer activities. However, the underlying mechanisms of myricetin on osteogenic differentiation remain unknown in human periodontal ligament stem cells (hPDLSCs). In this study, we investigated the ability of myricetin to increase osteogenic differentiation and its underlying molecular mechanisms. Myricetin significantly increased cell proliferation, alkaline phosphatase (ALP) activity, and alizarin red-mineralization activity in hPDLSCs in a dose-dependent manner. Furthermore, myricetin dose-dependently increased osteogenic-related mRNA and protein levels. Interestingly, it enhanced osteogenesis by up-regulating bone morphogenetic protein-2 (BMP-2), which induced the expression of BMP receptor type IB, Smad-1/5/9. It also enhanced the phosphorylation of extracellular signal-regulated kinases (ERKs), c-Jun N-terminal kinase (JNK), p38 mitogen-activated protein kinases (MAPKs) and Smads. We confirmed that the treatment of myricetin increased phosphorylated GSK-3β and β-catenin which is related to osteogenesis. In our studies, myricetin-induced increment of ALP activity was decreased by ERK (PD98059), JNK (SP600125), p38 (SB203580), and Smad 1/5/9 (LDN193189) inhibitors. ERK and p38 inhibitors showed the greatest inhibition among the four kinds of inhibitors. These results demonstrate that myricetin promoted osteogenic differentiation by the up-regulation of ALP activity and expression of osteogenic-related factors through BMP-2/Smad and ERK/JNK/p38 MAPK pathways. 10.1016/j.ejphar.2018.07.012
    Rutin protects human periodontal ligament stem cells from TNF-α induced damage to osteogenic differentiation through suppressing mTOR signaling pathway in inflammatory environment. Zhao Bin,Zhang Wenjing,Xiong Yixuan,Zhang Yunpeng,Jia Linglu,Xu Xin Archives of oral biology OBJECTIVES:To investigate whether rutin could protect human periodontal ligament stem cells (hPDLSCs) from TNF-α induced damage to osteogenic differentiation in inflammatory environment and detect the underlying mechanism. MATERIALS AND METHODS:hPDLSCs were identified by flow cytometery. TNF-α was used to stimulate hPDLSCs to establish an inflammation model in vitro. Alkaline phosphatase (ALP) staining, ALP activity test, and Alizarin Red staining were used to detect the changes of osteogenic differentiation ability. The mRNA and protein levels of osteogenic genes were evaluated by RT-PCR and Western Blot. The expression of mTOR was also detected by Western Blot. RESULTS:hPDLSCs were positive to MSCs specific surface markers. The inflammatory environment in vitro could be established by stimulating hPDLSCs with TNF-α (20 ng/mL). TNF-α (20 ng/mL) could decrease the ALP activity and mineralization ability of hPDLSCs and down-regulate the expression of osteogenic genes in inflammatory environment. Moreover, rutin could affect TNF-α (20 ng/mL) induced damage to osteogenic differentiation of hPDLSCs in a dose-dependent manner, 10 μmol/L rutin could significantly reverse the damage caused by TNF-α. In addition, rutin inhibited TNF-α-activated mTOR signal transduction by inhibiting the phosphorylation of mTOR, similar to the effects of rapamycin(a specific mTOR inhibitor). CONCLUSIONS:Rutin could protect hPDLSCs from TNF-α induced damage to osteogenic differentiation in inflammatory environment, and rutin is expected to become a new candidate drug for the treatment of bone defect of periodontitis. 10.1016/j.archoralbio.2019.104584
    Anti-inflammatory effects of isorhamnetin on LPS-stimulated human gingival fibroblasts by activating Nrf2 signaling pathway. Qi Feng,Sun Ji-Hao,Yan Jia-Qun,Li Chun-Mei,Lv Xue-Chao Microbial pathogenesis Periodontitis is a highly prevalent infective and inflammatory disease with an adverse impact on systemic health. Isorhamnetin, a flavonoid mainly isolated from Hippophae fhamnoides L. fruit, has been reported to have anti-inflammatory effect. This study aimed to investigate the anti-inflammatory effects and mechanism of isorhamnetin on lipopolysaccharide (LPS)-induced inflammatory response in human gingival fibroblasts (HGFs). The production of inflammatory mediators and the expression of proteins were measured by ELISA and western blot analysis. The results demonstrated that isorhamnetin attenuated LPS-induced release of PGE, NO, IL-6, and IL-8 in HGFs. Isorhamnetin also inhibited LPS-induced NF-κB activation. The expression of Nrf2 and HO-1 were up-regulated by treatment of isorhamnetin. Furthermore, knockdown of Nrf2 by siRNA reversed the anti-inflammatory effects of isorhamnetin. In conclusion, these results suggested that isorhamnetin inhibited LPS-induced inflammation in HGFs by activating Nrf2 signaling pathway. 10.1016/j.micpath.2018.04.049
    Quercetin reverses TNF‑α induced osteogenic damage to human periodontal ligament stem cells by suppressing the NF‑κB/NLRP3 inflammasome pathway. Zhang Wenjing,Jia Linglu,Zhao Bin,Xiong Yixuan,Wang Ya-Nan,Liang Jin,Xu Xin International journal of molecular medicine Quercetin (Quer) is a typical antioxidant flavonoid from plants that is involved in bone metabolism, as well as in the progression of inflammatory diseases. Elevated levels of tumor necrosis factor‑α (TNF‑α), a typical pro‑inflammatory cytokine, can affect osteogenesis. In the present study, TNF‑α was used to establish an model of periodontitis. The effects of Quer on, as well as its potential role in the osteogenic response of human periodontal ligament stem cells (hPDLSCs) under TNF‑α‑induced inflammatory conditions and the underlying mechanisms were then investigated. Within the appropriate concentration range, Quer did not exhibit any cytotoxicity. More importantly, Quer significantly attenuated the TNF‑α induced the suppression of osteogenesis‑related genes and proteins, alkaline phosphatase (ALP) activity and mineralized matrix in the hPDLSCs. These findings were associated with the fact that Quer inhibited the activation of the NF‑κB signaling pathway, as well as the expression of NLRP3 inflammation‑associated proteins in the inflammatory microenvironment. Moreover, the silencing of NLRP3 by small interfering RNA (siRNA) was found to protect the hPDLSCs against TNF‑α‑induced osteogenic damage, which was in accordance with the effects of Quer. On the whole, the present study demonstrates that Quer reduces the impaired osteogenesis of hPDLSCs under TNF‑α‑induced inflammatory conditions by inhibiting the NF‑κB/NLRP3 inflammasome pathway. Thus, Quer may prove to be a potential remedy against periodontal bone defects. 10.3892/ijmm.2021.4872
    DYNAMICS OF CHANGES OF C-REACTIVE PROTEIN LEVEL IN BLOOD SERUM IN THE DEVELOPMENT AND COURSE OF EXPERIMENTAL PERIODONTITIS AND THEIR CORRECTION BY FLAVONOL. Demkovych Andrii,Hasiuk Petro,Korobeinikova Yuliia,Shcherba Vitaliy,Korobeinikov Leonid Wiadomosci lekarskie (Warsaw, Poland : 1960) OBJECTIVE:The aim: To study the value of C-reactive protein in the experimental animals blood serum with bacterial-immune periodontitis and its correction with quercetin. PATIENTS AND METHODS:Materials and methods: Modeling of periodontitis was performed by the following method: after thiopental anesthesia (at a dose of 40 mg / kg intramuscularly) rats were fixed. A subcostal injection of 0.01 ml of egg protein with cultures of Streptococcus hemolytic and Staphylococcus aureus at a dose of 4 CFU was performed in the area of periodontal tissues of the lower incisor as an initiating inflammatory factor. To enhance the immune process, a complete Freund's adjuvant was introduced into the animal's hind limb at the same time. RESULTS:Results: Analysis of the results of the study of the content of C-reactive protein in the blood serum of animals with experimental bacteria and immune periodontitis, receiving injections of quercetin, showed a significant decrease by 1.31 times, compared with animals with this simulated pathology on the 14th day of the experiment without the use of flavonol. When comparing this indicator on the 14th day of development of experimental periodontitis with correction, it was found that it remained slightly higher than the indicators of the intact group of rats. CONCLUSION:Conclusions: The level of C-reactive protein in the blood serum of experimental animals is an important indicator of the immune-inflammatory response, which increases its activation of the inflammatory system. The administration of flavonoid quercetin for 7 days helps to reduce the level of C-reactive protein in the blood serum of animals with experimental bacterial and immune periodontitis.
    Dalbergiones lower the inflammatory response in oral cells in vitro. Clinical oral investigations OBJECTIVES:Periodontitis is a global health burden that underlines the demand for anti-inflammatory treatment. Dalbergia melanoxylon being a rich source of flavonoids has been widely used in traditional medicine but the potential anti-inflammatory activity of its dalbergiones remains to be shown. MATERIAL AND METHODS:We have isolated 3'-hydroxy-4,4'-dimethoxydalbergione, 4-methoxydalbergione, and 4'-hydroxy-4-methoxydalbergione from Dalbergia melanoxylon and tested their potential anti-inflammatory activity. RESULTS:All dalbergiones are potent inhibitors of an LPS-induced inflammatory response of RAW 264.7 macrophages. This is specified by IL1β and IL6 production, and the p65 nuclear translocation. Consistently, in primary macrophages, the dalbergiones caused an M1-to-M2 polarization switch indicated by the decreased ration of IL1β and IL6 versus arginase 1 and YM1 expression. To implement oral cells, we have used gingival fibroblasts exposed to IL1β and TNFα. Consistently, all dalbergiones reduced the expression of IL6 and IL8 as well as the nuclear translocation of p65. CONCLUSION:These findings increase the accumulating knowledge on dalbergiones and extend it towards its capacity to lower the inflammatory response of oral cells. CLINICAL RELEVANCE:These findings are another piece of evidence that supports the use of herbal medicine to potentially lower inflammatory events related to dentistry. 10.1007/s00784-022-04509-7
    Cyanidin-3-O-glucoside downregulates ligation-activated endoplasmic reticulum stress and alleviates induced periodontal destruction in rats. Tu Hsiao-Pei,Kuo Chan-Yen,Fu Martin Ming-Jen,Chin Yu-Tang,Chiang Cheng-Yang,Chiu Hsien-Chung,Hsia Yi-Jan,Fu Earl Archives of oral biology OBJECTIVE:Cyanidin-3-O-glucoside (C3G), a dietary anthocyanin, possesses various biological properties, including alleviating endoplasmic reticulum (ER) stress. This study examined the effect of C3G on periodontitis via ER stress in rats. DESIGN:Periodontitis was induced by placing silk sutures around maxillary second molars. C3G (0, 3, or 9 mg/kg) was fed on the day before ligation (10 rats/group). Further, 10 non-ligation control rats received deionized water. On day 8, gingivae were obtained to determine CCAAT/enhancer-binding protein homologous protein (CHOP), c-Jun N-terminal kinase (JNK), phospho-JNK (p-JNK), and nuclear factor-κB (NF-κB) by immunoblotting. Periodontal destruction was evaluated using micro-computed tomography (μCT) and histology. RESULTS:Gingival expression of CHOP, p-JNK/JNK, and NF-κB significantly increased in ligation rats (0 mg/kg C3G) than that in controls. However, protein expression in ligation groups presented a negative association with C3G concentration. By μCT, the distance of cemento-enamel junction to bone significantly increased in ligation groups; however, distances showed a negative association with C3G concentration. In the region of interest, bone volume and trabecular thickness and number significantly decreased in ligation groups but they were positively associated with C3G concentration. In terms of trabecular separation, opposite results were found. Histologically, infiltrated connective tissue (ICT) and periodontal destructions increased in ligation groups; however, they were negatively associated with C3G concentration. Moreover, ICT area is positively correlated with μCT- and histologically measured destructions and protein expression of CHOP, p-JNK/JNK, or NF-κB. CONCLUSION:C3G promotes favorable modulation of ER stress and alleviates destruction of periodontitis, which may imply a new strategy. 10.1016/j.archoralbio.2021.105313
    Tectorigenin Promotes Osteoblast Differentiation and Bone Healing, but Suppresses Osteoclast Differentiation and Bone Resorption. Lee So-Youn,Kim Gyu-Tae,Yun Hyung-Mun,Kim Youn-Chul,Kwon Il-Keun,Kim Eun-Cheol Molecules and cells Although tectorigenin (TG), a major compound in the rhizome of , is conventionally used for the treatment of inflammatory diseases, its effects on osteogenesis and osteoclastogenesis have not been reported. The objective of this study was to investigate the effects and possible underlying mechanism of TG on osteoblastic differentiation and bone formation, as well as osteoclast differentiation and bone resorption. TG promoted the osteogenic differentiation of primary osteoblasts and periodontal ligament cells. Moreover, TG upregulated the expression of the BMP2, BMP4, and Smad-4 genes, and enhanced the expression of Runx2 and Osterix. studies involving mouse calvarial bone defects with μCT and histologic analysis revealed that TG significantly increased new bone formation. Furthermore, TG treatment inhibited osteoclast differentiation and the mRNA levels of osteoclast markers. studies of mice demonstrated that TG caused the marked attenuation of bone resorption. These results collectively demonstrated that TG stimulated osteogenic differentiation , increased bone regeneration, inhibited osteoclast differentiation , and suppressed inflammatory bone loss . These novel findings suggest that TG may be useful for bone regeneration and treatment of bone diseases. 10.14348/molcells.2018.0056
    Baicalein enhances the osteogenic differentiation of human periodontal ligament cells by activating the Wnt/β-catenin signaling pathway. Chen Li-Jiao,Hu Bi-Bo,Shi Xin-Lian,Ren Man-Man,Yu Wen-Bin,Cen Sheng-Dan,Hu Rong-Dang,Deng Hui Archives of oral biology OBJECTIVE:Periodontium regeneration is one of the most important processes for periodontitis therapy. Human periodontal ligament cells (hPDLCs) play a vital role in the repair and regeneration of periodontal tissues. Our study aimed to investigated the mechanisms underlying the promotion of hPLDCs osteogenic differentiation by baicalein. DESIGN:hPDLCs were obtained from periodontal ligament (PDL) tissues by primary culture. The MTT assay was used to determine the growth curves of hPDLCs treated with different concentrations of baicalein (1.25, 2.5, 5, or 10μM). Alkaline phosphatase (ALP) staining and Alizarin red S staining were performed to assess osteogenic differentiation of hPDLCs administered baicalein. Osteogenic differentiation-related gene and protein expression levels and Wnt/β-catenin pathway signal changes were assessed by qRT-PCR and Western blotting analysis. RESULTS:The results showed that baicalein decreased the growth of hPDLCs slightly and increased ALP activity and calcium deposition in a dose-dependent manner. The expression of runt-related transcription factor 2 (RUNX2), bone morphogenetic protein 2 (BMP2), Osterix (OSX) and osteocalcin (OCN) were elevated after baicalein administration. Moreover, baicalein strongly activated the Wnt/β-catenin pathway and up-regulated the expression of β-catenin, lymphoid enhancer factor 1 (LEF1) and Cyclin D1. Dickkopf-related protein 1 (DKK-1) significantly reversed the effects of baicalein on hPDLCs. CONCLUSIONS:Our findings indicated that baicalein enhanced the osteogenic differentiation of hPDLCs via the activation of the Wnt/β-catenin signaling pathway, which may represent a potential candidate for periodontitis therapy. 10.1016/j.archoralbio.2017.01.019
    Sudachitin Inhibits Matrix Metalloproteinase-1 and -3 Production in Tumor Necrosis Factor-α-Stimulated Human Periodontal Ligament Cells. Hosokawa Yoshitaka,Hosokawa Ikuko,Ozaki Kazumi,Matsuo Takashi Inflammation Sudachitin, a polymethoxylated flavonoid found in the skin of Citrus sudachi, is a biologically active substance. The aim of this study was to examine whether sudachitin could be used to inhibit the expression of matrix metalloproteinase (MMP)-1 and MMP-3, which are involved in the destruction of periodontal tissues in periodontal lesions, in tumor necrosis factor (TNF)-α-stimulated human periodontal ligament cells (HPDLC). Sudachitin suppressed TNF-α-induced MMP-1 and MMP-3 production in HPDLC. On the other hand, it enhanced tissue inhibitor of metalloproteinase (TIMP)-1 expression. The level of Akt phosphorylation in the TNF-α-stimulated HPDLC was decreased by sudachitin treatment. Moreover, an Akt inhibitor reduced MMP-1 and MMP-3 production and increased TIMP-1 production. These findings indicate that sudachitin reduces MMP-1 and MMP-3 production in TNF-α-stimulated HPDLC by inhibiting the Akt pathway. 10.1007/s10753-019-01007-z
    Effects of taxifolin on bone formation and apoptosis in experimental periodontitis in diabetic rats. Lektemur Alpan Aysan,Bakar Olcay,Kızıldağ Alper,Özdede Melih,Topsakal Şenay,Özmen Özlem Biotechnic & histochemistry : official publication of the Biological Stain Commission We investigated the therapeutic potential of taxifolin for treatment of alveolar bone loss (ABL) in experimental periodontitis in diabetic rats. Diabetes mellitus (DM) was induced by streptozotocin. Rats were divided into six groups: untreated control; DM only (D) group; ligature only (P) group; DM + ligature (DP) group; DM + ligature + 5 mg/kg/day taxifolin (Taxi-5) group; DM + ligature + 10 mg/kg/day taxifolin (Taxi-10) group. Experimental periodontitis was induced by ligation of the first molar and allowed to progress for 30 days before performing cone-beam computed tomographic (CBCT), histomorphometric and immunohistochemical analyses of periodontal tissue destruction. ABL was assessed using CBCT. ABL was greatest in the P and DP groups. Decreased ABL was observed in the Taxi-5 and Taxi-10 groups. Bone morphogenic protein (BMP-2), osteocalcin (OCN), receptor activator of nuclear factor kappa-Β ligand (RANKL), alkaline phosphatase (ALP), type I collagen, B cell lymphoma-associated X (Bax), and B-cell lymphoma 2 (Bcl-2) levels were investigated using immunohistochemistry. The Taxi-5 and Taxi-10 groups exhibited decreased RANKL expression, but increased BMP-2, ALP, type I collagen and OCN levels compared to the P and DP groups. Bax activity was increased in the D, P and DP groups. Taxi-5 and Taxi-10 groups exhibited increased Bcl-2 activity. Our findings suggest that taxifolin can reduce apoptosis and improve alveolar bone formation in diabetic rats with periodontitis. 10.1080/10520295.2021.1977997
    A semi-synthetic flavonoid from Bauhinia pulchella stem attenuates inflammatory osteolysis in periodontitis in rats: Impact on cytokine levels, oxidative stress, and RANK/RANKL/OPG pathway. Pinto Isabela R,Chaves Hellíada V,Freire Jordânia M O,de Sousa Luzia Herminia T,Monteiro Dina A M,Costa José Jackson do N,Pereira Karuza M A,Santiago Gilvandete Maria P,de Sousa Leôncio M,da Silva Marcos Reinaldo,Monteiro Aurélio de O,Montenegro Raquel de C,de Moraes Maria Elisabete A,Filho Gerardo C,Pinto Vicente de Paulo T,Bezerra Mirna M Archives of oral biology OBJECTIVES:Many species of theBauhinia genus have been widely used in folk medicine as analgesic, anti-inflammatory and antioxidant agents. (-)-Fisetinidol palmitate is a semi-syntetic flavonoid obtained from the ethanolic extract of the stem of Bauhinia pulchella. This study aimed to evaluate the antiresorptive effect of the semi-syntetic (-)-fisetinidol palmitate in ligature-induced periodontitis in rats. Also, it evaluated the mechanism of action of (-)-fisetinidol palmitate and its toxicity. DESIGN:Periodontitis was inducedvia a nylon thread ligature (3.0) around the second upper left molars. Rats were treated (oral gavage) once a day for 11 days with (-)-fisetinidol palmitate (0.01 or 0.1 mg/kg) or saline vehicle. RESULTS:(-)-Fisetinidol palmitate (0.1 mg/kg) reduced alveolar bone loss, increased bone alkaline phosphatase (BALP), superoxide dismutase (SOD), and catalase (CAT) activity; also, it decreased IL1-β, IL-8/CINC-1, nitrite/nitrate levels and myeloperoxidase activity. (-)-Fisetinidol palmitate reduced the mRNA levels of IL1-β, IL-6, RANK, and RANK-L, while it increased the OPG ones. No statistical differences (P > 0.05) were observed in the transaminases (ALT, AST) and Total Alkaline Phosphatase (TALP) levels among groups. (-)- CONCLUSIONS:Fisetinidol palmitate did not result in any signs of toxicity and had anti-resorptive effects in a pre-clinical trial of periodontitis, showing antioxidant activity with the involvement of the RANK/RANKL/OPG pathway. 10.1016/j.archoralbio.2020.104816
    Rutin promotes the formation and osteogenic differentiation of human periodontal ligament stem cell sheets in vitro. Zhao Bin,Zhang Yunpeng,Xiong Yixuan,Xu Xin International journal of molecular medicine Cell sheet technology is a novel tissue engineering technology that has been rapidly developed in recent years. As a novel technology, cell sheet technology is expected to become one of the preferred methods for cell transplantation. The present study investigated the biological effects of rutin on the formation of periodontal ligament stem cell (PDLSC) sheets and their resultant osteogenic properties. The results of Cell Counting Kit‑8 (CCK‑8) assay demonstrated that a concentration of 1x10‑6 mol/l rutin promoted the proliferation of PDLSCs more effectively compared with other designed concentrations. Rutin‑modified cell sheets could be induced by complete medium supplemented with 20 µg/ml vitamin C (VC) and 1x10‑6 mol/l rutin. Rutin‑modified cell sheets appeared thicker and more compact compared with the VC‑induced PDLSC sheets, demonstrating more layers of cells (3 or 4 layers), which secreted a richer extracellular matrix (ECM). Furthermore, the improved cell sheets exhibited varying degrees of increases in the mRNA and protein expression of collagen type I (COL1), alkaline phosphatase (ALP), runt‑related transcription factor 2 (RUNX2) and osteopontin (OPN). Combined treatment with VC and rutin promoted the formation of PDLSC sheets and enhanced the osteogenic differentiation potential of the cell sheets. Therefore, rutin‑modified cell sheets of PDLSCs are expected to play an important role in the treatment of periodontal tissue regeneration by stem cells. 10.3892/ijmm.2019.4384
    Baicalin Promotes Osteogenic Differentiation of Human Cementoblast Lineage Cells Via the Wnt/β Catenin Signaling Pathway. Kimura Aya,Kunimatsu Ryo,Yoshimi Yuki,Tsuka Yuji,Awada Tetsuya,Horie Kayo,Gunji Hidemi,Abe Takaharu,Nakajima Kengo,Kitagawa Masae,Miyauchi Mutsumi,Takata Takashi,Tanimoto Kotaro Current pharmaceutical design BACKGROUND:Baicalin constitutes a natural bioactive flavonoid extracted from Scutellaria baicalensis Georgi that mediates bone formation. However, the biological functions of baicalin in cementoblasts remain unclear. The purpose of this study was to examine the effects of baicalin on osteogenic differentiation of human cementoblast (HCEM) cells. METHODS:HCEM cells were cultured and treated with 0, 0.01, 0.1 or 1 µM baicalin. Alkaline phosphatase (ALP) and runt-related transcription factor 2 (Runx2) mRNA and protein levels were examined by real-time polymerase chain reaction and western blot analysis, respectively. Cell mineralization was assessed using Alizarin red staining. Glycogen synthase kinase-3 beta (GSK3β) phosphorylation was measured in 1 µM baicalin-treated HCEM cells with or without the Wnt signaling pathway inhibitor, DKK-1 using ELISA and western blotting. RESULTS:The protein levels of ALP and Runx2 and the intensity of Alizarin red staining were enhanced by baicalin in a dose-dependent manner compared to that of the non-treated control. The ratio of phosphorylated to total GSK3β increased in the presence of baicalin but was reduced by the addition of DKK-1. Treatment of HCEMs with baicalin up-regulated mRNA levels of ALP and Runx2, which were reduced by DKK-1. In addition, the protein levels of ALP and Runx2, ALP activity, and calcium deposition were also enhanced by baicalin, and these parameters were inhibited by DKK-1. CONCLUSION:Baicalin enhanced osteogenic differentiation of HCEM cells through the Wnt/beta catenin signaling pathway which may be useful for periodontal tissue regeneration. 10.2174/1381612824666181116103514
    Inhibitory Effects of Panduratin A on Periodontitis-Induced Inflammation and Osteoclastogenesis through Inhibition of MAPK Pathways In Vitro. Kim Haebom,Kim Mi-Bo,Kim Changhee,Hwang Jae-Kwan Journal of microbiology and biotechnology Periodontitis is an inflammatory disease caused by microbial lipopolysaccharide (LPS), destroying gingival tissues and alveolar bone in the periodontium. In the present study, we evaluated the anti-inflammatory and anti-osteoclastic effects of panduratin A, a chalcone compound isolated from , in human gingival fibroblast-1 (HGF-1) and RAW 264.7 cells. Treatment of panduratin A to LPS-stimulated HGF-1 significantly reduced the expression of interleukin-1β and nuclear factor-kappa B (NF-κB), subsequently leading to the inhibition of matrix metalloproteinase-2 (MMP-2) and MMP-8 compared with that in the LPS control (** < 0.01). These anti-inflammatory responses were mediated by suppressing the mitogen-activated protein kinase (MAPK) signaling and activator protein-1 complex formation pathways. Moreover, receptor activator of NF-κB ligand (RANKL)-stimulated RAW 264.7 cells treated with panduratin A showed significant inhibition of osteoclastic transcription factors such as nuclear factor of activated T-cells c1 and c-Fos as well as osteoclastic enzymes such as tartrate-resistant acid phosphatase and cathepsin K compared with those in the RANKL control (** < 0.01). Similar to HGF-1, panduratin A suppressed osteoclastogenesis by controlling MAPK signaling pathways. Taken together, these results suggest that panduratin A could be a potential candidate for development as a natural anti-periodontitis agent. 10.4014/jmb.1707.07042
    Anti-inflammatory effect of paeoniflorin combined with baicalin in oral inflammatory diseases. Yu Feiyan,Xu Na,Zhou Yu,Li Baoyin,Li Miao,Wang Qianqian,Yang Xi,Ge Xuejun,Zhang Fang,Ren Xiuyun Oral diseases OBJECTIVE:There are challenges in the treatment of chronic inflammatory diseases of oral mucosa. Both paeoniflorin (PF) and baicalin (BAI) exert anti-inflammatory effects, but the mechanism underlying their combined effects is still unclear. Here, we explored the anti-inflammatory function of the PF-BAI combination in the oral inflammatory response. MATERIALS AND METHODS:The CCK-8 assay was used to determine the proliferative capacity of HOKs with PF and BAI. Enzyme-linked immunosorbent (ELISA), Western blotting, reverse transcription polymerase chain reaction, and confocal immunofluorescence were performed to study the anti-inflammatory effects of PF-BAI in LPS-stimulated human oral keratinocytes (HOKs). Immunohistochemistry and ELISA were performed to detect the levels of NF-κB p65, IKKα and IL-6, TNF-α in OLP and healthy tissues. RESULTS:Compared to PF or BAI alone, the combination of PF-BAI at 5 µg/ml downregulated secretion of inflammatory cytokines more effectively (p < .05). Combined PF-BAI decreased NF-κB p65 and IκBα protein phosphorylation, leading to reduce nuclear translocation of NF-κB p65. Higher expression of TNF-α, IL-6, NF-κB p65, and IKKα were observed in OLP than in HC tissues (p < .01). CONCLUSION:The optimal combination concentration of PF and BAI at 5 µg/ml may have a positive effect on the treatment of oral inflammatory diseases, providing a novel therapeutic approach. 10.1111/odi.13171
    Identification of Interleukin-8-Reducing Lead Compounds Based on SAR Studies on Dihydrochalcone-Related Compounds in Human Gingival Fibroblasts (HGF-1 cells) In Vitro. Schueller Katharina,Hans Joachim,Pfeiffer Stefanie,Walker Jessica,Ley Jakob P,Somoza Veronika Molecules (Basel, Switzerland) In order to identify potential activities against periodontal diseases, eighteen dihydrochalcones and structurally related compounds were tested in an established biological in vitro cell model of periodontal inflammation using human gingival fibroblasts (HGF-1 cells). Methods: Subsequently to co-incubation of HGF-1 cells with a bacterial endotoxin ( lipopolysaccharide, LPS) and each individual dihydrochalcone in a concentration range of 1 µM to 100 µM, gene expression of interleukin-8 (IL-8) was determined by qPCR and cellular interleukin-8 (IL-8) release by ELISA. : Structure-activity analysis based on the dihydrochalcone backbone and various substitution patterns at its aromatic ring revealed moieties 2',4,4',6'-tetrahydroxy 3-methoxydihydrochalcone () to be the most effective anti-inflammatory compound, reducing the LPS-induced IL-8 release concentration between 1 µM and 100 µM up to 94%. In general, a 2,4,6-trihydroxy substitution at the A-ring and concomitant vanilloyl (4-hydroxy-3-methoxy) pattern at the B-ring revealed to be preferable for IL-8 release inhibition. Furthermore, the introduction of an electronegative atom in the A,B-linker chain led to an increased anti-inflammatory activity, shown by the potency of 4-hydroxybenzoic acid -vanillylamide (). Our data may be feasible to be used for further lead structure designs for the development of potent anti-inflammatory additives in oral care products. 10.3390/molecules25061382
    Systemic Dietary Hesperidin Modulation of Osteoclastogenesis, Bone Homeostasis and Periodontal Disease in Mice. International journal of molecular sciences This study aimed to evaluate the effects of hesperidin (HE) on in vitro osteoclastogenesis and dietary supplementation on mouse periodontal disease and femoral bone phenotype. RAW 264.7 cells were stimulated with RANKL in the presence or absence of HE (1, 100 or 500 µM) for 5 days, and evaluated by TRAP, TUNEL and Western Blot (WB) analyses. In vivo, C57BL/6 mice were given HE via oral gavage (125, 250 and 500 mg/kg) for 4 weeks. A sterile silk ligature was placed between the first and second right maxillary molars for 10 days and microcomputed tomography (μCT), histopathological and immunohistochemical evaluation were performed. Femoral bones subjected or not to dietary HE (500 mg/kg) for 6 and 12 weeks were evaluated using μCT. In vitro, HE 500 µM reduced formation of RANKL-stimulated TRAP-positive(+) multinucleated cells (500 µM) as well as c-Fos and NFATc1 protein expression ( &lt; 0.05), markers of osteoclasts. In vivo, dietary HE 500 mg/kg increased the alveolar bone resorption in ligated teeth ( &lt; 0.05) and resulted in a significant increase in TRAP+ cells ( &lt; 0.05). Gingival inflammatory infiltrate was greater in the HE 500 mg/kg group even in the absence of ligature. In femurs, HE 500 mg/kg protected trabecular and cortical bone mass at 6 weeks of treatment. In conclusion, HE impaired in vitro osteoclastogenesis, but on the contrary, oral administration of a high concentration of dietary HE increased osteoclast numbers and promoted inflammation-induced alveolar bone loss. However, HE at 500 mg/kg can promote a bone-sparing effect on skeletal bone under physiological conditions. 10.3390/ijms23137100
    The Polymethoxy Flavonoid Sudachitin Inhibits Interleukin-1-Induced Inflammatory Mediator Production in Human Periodontal Ligament Cells. Hosokawa Yoshitaka,Hosokawa Ikuko,Ozaki Kazumi,Matsuo Takashi BioMed research international Sudachitin, which is a polymethoxylated flavonoid found in the peel of Citrus sudachi, has some biological activities. However, the effect of sudachitin on periodontal resident cells is still uncertain. The aim of this study was to examine if sudachitin could decrease the expression of inflammatory mediators such as cytokines, chemokines, or matrix metalloproteinase (MMP) in interleukin- (IL-) 1-stimulated human periodontal ligament cells (HPDLC). Sudachitin inhibited IL-1-induced IL-6, IL-8, CXC chemokine ligand (CXCL)10, CC chemokine ligand (CCL)2, MMP-1, and MMP-3 production in HPDLC. On the other hand, tissue inhibitor of metalloproteinase- (TIMP-) 1 expression was increased by sudachitin treatment. Moreover, we found that the nuclear factor- (NF-) B and protein kinase B (Akt) pathways in the IL-1-stimulated HPDLC were inhibited by sudachitin treatment. These findings indicate that sudachitin is able to reduce inflammatory mediator production in IL-1-stimulated HPDLC by inhibiting NF-B and Akt pathways. 10.1155/2021/8826586
    Hyperoside ameliorates periodontitis in rats by promoting osteogenic differentiation of BMSCs via activation of the NF-κB pathway. Xu Tao,Wu Xiao,Zhou Zhou,Ye Yu,Yan Chaoting,Zhuge Nanshan,Yu Jinhua FEBS open bio Hyperoside, as an active compound, widely exists in a large number of Chinese herbal medicines and has been reported to possess anti-inflammatory and diuretic properties. However, the effects and underlying mechanisms of hyperoside on periodontitis have not been previously reported. In this study, we found that hyperoside ameliorates symptoms of periodontitis in a rat model, with improvements in alveolar bone resorption, relief of inflammatory infiltration, increase in orderly arrangement of collagen fibers and increase of osteogenic differentiation. In addition, hyperoside promoted proliferation, up-regulated EdU-positive cells, decreased cell-cycle distribution and increased the protein expression of Ki67 and PCNA in rat bone mesenchymal stem cells (rBMSCs), as revealed by Cell Counting Kit-8, EdU, flow cytometry and western blot analysis. Moreover, hyperoside significantly promoted osteogenic differentiation, as shown by quantitative RT-PCR, western blot and alizarin red staining assays. Furthermore, hyperoside activated the nuclear factor-κB (NF-κB) signaling pathway in rBMSCs, similar to the results observed in vivo. Finally, BMS345541, an inhibitor of the NF-κB signaling pathway, could reverse the effects of hyperoside on the biological functions in rBMSCs. In conclusion, our results suggest that hyperoside has potential therapeutic properties against periodontitis via promotion of proliferation and osteogenic differentiation of rBMSCs via activation of the NF-κB signaling pathway. 10.1002/2211-5463.12937
    Effect of puerarin on osteogenic differentiation of human periodontal ligament stem cells. Li Jun,Peng Youjian The Journal of international medical research OBJECTIVE:To investigate the effects of the flavonoid, puerarin, on osteogenic differentiation of human periodontal ligament stem cells (PDLSCs). METHODS:Human PDLSCs were isolated from patients undergoing orthodontic treatment, and the cell surface markers CD146, CD34, CD45, and STRO-1 were identified by immunofluorescence. Cell proliferation was detected by MTT assay; alkaline phosphatase (ALP) activity was measured, and calcium deposition was detected by alizarin red staining. PCR was then used to detect the distributions of , , , and , genes related to osteogenic differentiation. RESULTS:Staining was positive for cytokines CD146, CD34, CD45, and STRO-1 in the experimental group; staining was also positive for silk protein, but negative for keratin. After 7 days of culture, exposure to puerarin significantly promoted the level of intracellular ALP; increased puerarin concentration led to increased intracellular ALP. Red mineralized nodules appeared upon exposure to puerarin and the number of nodules was concentration-dependent. PCR analysis revealed that , , , and expression levels increased as puerarin concentration increased. CONCLUSIONS:Exposure to puerarin can promote proliferation and ALP activity in human PDLSCs, thus promoting both molecular and osteogenic differentiation; these findings may provide a theoretical basis for the clinical treatment of periodontal disease with puerarin. 10.1177/0300060519851641
    Nobiletin Decreases Inflammatory Mediator Expression in Tumor Necrosis Factor-Stimulated Human Periodontal Ligament Cells. Hosokawa Yoshitaka,Hosokawa Ikuko,Ozaki Kazumi Mediators of inflammation Nobiletin, a biologically active substance in the skin of citrus fruits, has been reported to be an effective anti-inflammatory, anticancer, and antimicrobial agent. In this study, we aimed to examine the anti-inflammatory effects of nobiletin on tumor necrosis factor- (TNF-) stimulated human periodontal ligament cells (HPDLCs). Our results demonstrated that nobiletin treatment could decrease the expressions of inflammatory cytokines (C-X-C motif chemokine ligand (CXCL)10, C-C motif chemokine ligand (CCL)2, and interleukin- (IL-) 8), matrix metalloproteinases (MMPs) (MMP1 and MMP3), and prostaglandin-endoperoxide synthase 2 (PTGS2) in TNF-stimulated HPDLCs. Moreover, we revealed that nobiletin could inhibit the activation of nuclear factor- (NF-) B and protein kinase B (AKT1) pathways in TNF-stimulated HPDLCs. Furthermore, nobiletin treatment enhanced nuclear factor, erythroid 2 like 2 (NFE2L2) and heme oxygenase 1 (HMOX1) expressions in TNF-stimulated HPDLCs. In conclusion, these findings suggest that nobiletin can inhibit inflammatory responses in TNF-stimulated HPDLCs by inhibiting NF-B and AKT1 activations and upregulating the NFE2L2 and HMOX1 expression. 10.1155/2021/5535844
    Protective effect of acacetin in human periodontal ligament cells regulation of autophagy and inflammation. Liu Jia,Wang Yu-Guang,Yu Shu-Yan,Li Chun-E,Kang Si-Meng Die Pharmazie Our study investigated the effects of acacetin, a natural flavonoid compound, on the survival and expression of inflammatory related cytokines in lipopolysaccharide (LPS)-stimulated human periodontal ligament (PDL) cells. Treatment with acacetin significantly promoted survival and suppressed apoptosis in LPS-stimulated PDL cells in a dose-dependent manner, as shown by CCK-8 and flow cytometry assays, respectively. Moreover, ELISA assay showed that acacetin dose-dependently attenuated LPS-induced increases of TNF-α, IL-6 and IL-1β in PDL cells. Western blot analysis showed that administration of acacetin dose-dependently increased the ratio of LC3II/LC3I, as well as the expression of beclin-1, as compared to LPS-stimulated PDL cells. Inhibition of autophagy by rapamycin, an autophagy inhibitor, increased the production of pro-inflammatory cytokines and decreased survival, abolishing the beneficial role of acacetin in LPS-stimulated PDL cells. In addition, the expression of GSK-3β, a regulator of autophagy, was suppressed by administration with acacetin in a dose-dependent manner. Acacetin treatment promotes survival and suppresses inflammation in LPS-stimulated PDL cells via regulating autophagy and GSK-3β signal in PDL cells, suggesting that acacetin may be a potential novel agent for the treatment of chronic periodontitis. 10.1691/ph.2020.0435
    Influence of epigallocatechin-3-gallate in promoting proliferation and osteogenic differentiation of human periodontal ligament cells. Liu Jie,Lu Yi,Liu Jin,Jin Changxiong,Meng Yuchen,Pei Dandan BMC oral health BACKGROUND:Epigallocatechin-3-gallate (EGCG) was recently proposed to have the potential to regulate bone metabolism, however, its influence on osteogenesis remains controversial. The present study aimed to investigate the effects of EGCG on the proliferation and osteogenesis of human periodontal ligament cells (hPDLCs). METHODS:Cells were cultured in osteogenic medium and treated with EGCG at various concentrations. Cell proliferation was analyzed using a CCK-8 assay and acridine orange (AO)/ethidium bromide (EB) staining. Flow cytometry was used to measure the intracellular reactive oxygen species (ROS) potential of hPDLCs. The expression levels of osteogenic marker genes and proteins in hPDLCs, including type I collagen (COL1), runt-related transcription factor 2 (RUNX2), osteopontin (OPN), and osterix (OSX), were determined by quantitative real-time polymerase chain reaction (qRT-PCR) and western blot analysis. In addition, alkaline phosphatase (ALP) activity was monitored both quantitatively and qualitatively. Extracellular matrix mineralization was further analyzed by alizarin red S staining. RESULTS:The results showed that EGCG concentrations from 6 to 10 μM increased the ROS level and inhibited the cell proliferation of hPDLCs. EGCG concentrations from 2 to 8 μM effectively increased extracellular matrix mineralization, in which 4 and 6 μM EGCG generated the most mineralizing nodules. The ALP activity and the mRNA and protein expression levels of the tested osteogenic markers were most strongly up-regulated by treatment with 4 and 6 μM EGCG. CONCLUSIONS:The present study demonstrated that EGCG might promote the osteogenesis of hPDLCs in a dose-dependent manner, with concentrations of 4 and 6 μM EGCG showing the strongest osteogenic enhancement without cytotoxicity, indicating a promising role for EGCG in periodontal regeneration in patients with deficient alveolar bone in the future. 10.1186/s12903-019-0768-7
    Cynaroside protects human periodontal ligament cells from lipopolysaccharide-induced damage and inflammation through suppression of NF-κB activation. Lee Seul Ah,Park Bo-Ram,Moon Sung-Min,Shin Sang Hun,Kim Jae-Sung,Kim Do Kyung,Kim Chun Sung Archives of oral biology OBJECTIVE:To investigate whether cynaroside protects human periodontal ligament (hPDL) cells from lipopolysaccharide (LPS)-induced damage and inflammation and to analyze the underlying mechanism. METHODS:LPS was used to stimulate hPDL and RAW264.7 cells. MTT assay was used to detect cell viability, and protein expression levels were measured via western blot analysis. Nitrite oxide and prostaglandin E were used to quantify the inflammatory response. Alizarin Red S staining was used to detect mineralized nodules. RESULTS:Cynaroside inhibited the expression of iNOS, COX-2, TNF-α, and IL-6 in LPS-stimulated hPDL and RAW264.7 cells without cytotoxicity. Furthermore, cynaroside significantly suppressed LPS-induced protein expression of matrix metalloproteinase 3. Additionally, cynaroside prevented LPS-induced NF-κB p65 subunit translocation to the nucleus by inhibiting the phosphorylation and degradation of IκB-α. Moreover, cynaroside could restore the mineralization ability of hPDL cells reduced by LPS. CONCLUSION:Cynaroside protected hPDL cells from LPS-induced damage and inflammation via inhibition of NF-κB activation. These results suggest that cynaroside may be a potential therapeutic agent for the alleviation of periodontitis. 10.1016/j.archoralbio.2020.104944
    Luteolin supports osteogenic differentiation of human periodontal ligament cells. Quan He,Dai Xiaopeng,Liu Meiyan,Wu Chuanjun,Wang Dan BMC oral health BACKGROUND:Previous research revealed that luteolin could improve the activation of alkaline phosphatase (ALP) and osteocalcin in mouse osteoblasts. We aimed to determine the effect of luteolin on osteogenic differentiation of periodontal ligament cells (PDLCs). METHODS:Cultured human PDLCs (HPDLCs) were treated by luteolin at 0.01, 0.1, 1, 10, 100 μmol/L, Wnt/β-catenin pathway inhibitor (XAV939, 5 μmol/L) alone or in combination with 1 μmol/L luteolin. Immunohistochemical staining was performed to ensure cells source. Cell activity and the ability of osteogenic differentiation in HPDLCs were determined by MTT, ALP and Alizarin Red S staining. Real-time Quantitative PCR Detecting System (qPCR) and Western blot were performed to measure the expressions of osteogenic differentiation-related genes such as bone morphogenetic protein 2 (BMP2), osteocalcin (OCN), runt-related transcription factor 2 (RUNX2), Osterix (OSX) and Wnt/β-catenin pathway proteins members cyclin D1 and β-catenin. RESULTS:Luteolin at concentrations of 0.01, 0.1, 1, 10, 100 μmol/L promoted cell viability, ALP activity and increased calcified nodules content in HPDLCs. The expressions of BMP2, OCN, OSX, RUNX2, β-catenin and cyclin D1 were increased by luteolin at concentrations of 0.01, 0.1, 1 μmol/L, noticeably, 1 μmol/L luteolin produced the strongest effects. In addition, XAV939 inhibited the expressions of calcification and osteogenic differentiation-related genes in HPDLCs, and 1 μmol/L luteolin availably decreased the inhibitory effect. CONCLUSION:1 μmol/L luteolin accelerated osteogenic differentiation of HPDLCs via activating the Wnt/β-catenin pathway, which could be clinically applied to treat periodontal disease. 10.1186/s12903-019-0926-y
    Enhanced efficacy of baicalin-loaded TPGS polymeric micelles against periodontitis. Liu Xiaochen,Chen Yunong,Chen Xin,Su Jiansheng,Huang Chen Materials science & engineering. C, Materials for biological applications As a chronic infectious disease, periodontitis is the main cause of teeth exfoliation due to its severe inflammatory reaction and periodontal tissue destruction. Recent reports have shown that baicalin could inhibit the NF-κB signaling pathway in inflammatory activity of periodontitis, but the efficacy of baicalin is limited due to its poor water solubility. In this work, we report the fabrication and application of baicalin encapsulated D-α-tocopherol polyethylene glycol 1000 succinate (TPGS) polymeric micelles (PMs) through thin-film hydration method. The monodispersed micelles showed a spherical shape in aqueous solution and a prolonged drug-release kinetic. After baicalin was loaded into PMs, cytotoxicity and apoptosis induction were both decreased. The expression of genes (including TNF-α, IL-1β, RANKL and NF-κB) and the phosphorylation level of NF-κB p65 protein in lipopolysaccharide (LPS)-induced rat gingival fibroblasts were also reduced. Further investigation of drug efficacy in a rat periodontal disease model confirmed that the use of baicalin-PMs could reduce the destruction of alveolar bone and gingival fiber. Moreover, the therapeutic effect of baicalin-PMs was significantly better than that of free baicalin. These results suggest that the direct injection of micelles containing water-insoluble drugs may become a simple but effective method for treating periodontitis. 10.1016/j.msec.2019.03.103
    Rutin promotes osteogenic differentiation of periodontal ligament stem cells through the GPR30-mediated PI3K/AKT/mTOR signaling pathway. Zhao Bin,Xiong Yixuan,Zhang Yunpeng,Jia Linglu,Zhang Wenjing,Xu Xin Experimental biology and medicine (Maywood, N.J.) 10.1177/1535370220903463
    Ameliorative effect of hesperidin on ligation-induced periodontitis in rats. Kuo Po-Jan,Fu Earl,Lin Chi-Yu,Ku Cheng-Te,Chiang Cheng-Yang,Fu Martin Mj,Fu Min-Wen,Tu Hsiao-Pei,Chiu Hsien-Chung Journal of periodontology BACKGROUND:This study evaluated the ameliorative effect of hesperidin (HES), an anti-inflammatory flavanone, in rats with ligation (Lig)-induced periodontitis. METHODS:A total of 48 rats were randomly divided into non-ligation group (NL), Lig group, and two ligation-plus-HES groups (L+H). HES was administered immediately after ligature placement at a dose of 75 or 150 mg/kg by intragastric feeding. Destruction of the ligated maxillary second and mandibular first molars were evaluated by dental radiography, microcomputed tomography (micro-CT), and histometry performed after sacrificing the rats on the seventh day. The expression levels of interleukin (IL)-1β, IL-6, and inducible nitric oxide synthase (iNOS) messenger (m)RNAs in the gingiva were determined by reverse-transcription polymerase chain reaction. The expression of iNOS was examined by immunohistochemistry. RESULTS:The dental radiography and micro-CT findings revealed significantly increased alveolar bone loss in the Lig group, which was significantly prevented by HES. The histometry results revealed less gingival inflammation and connective tissue loss in the L+H groups compared with that in the Lig group. The mRNA expression levels of IL-6, IL-1 β, and iNOS were significantly increased in the Lig group but were reduced in the L+H groups. The immunostaining results showed that the ligation-induced iNOS expression was also decreased by HES. CONCLUSIONS:Oral administration of HES promotes an ameliorative effect against the ligation-induced alveolar bone loss and effectively inhibits the production of proinflammatory mediators in rats with experimentally induced periodontitis. Therefore, HES may be a good candidate for modulating oral inflammatory diseases. 10.1002/JPER.16-0708
    Nanoparticle-encapsulated baicalein markedly modulates pro-inflammatory response in gingival epithelial cells. Li Xuan,Luo Wei,Ng Tsz Wing,Leung Ping Chung,Zhang Chengfei,Leung Ken Cham-Fai,Jin Lijian Nanoscale Severe gum disease (periodontitis), which is one of the major global oral diseases, results from microbe-host dysbiosis and dysregulated immuno-inflammatory responses. It seriously affects oral health and general wellbeing with significant socio-economic implications. It has been well documented that natural flavonoids such as baicalin (BA) and baicalein (BE) possess potent anti-inflammatory effects. However, their intrinsic poor solubility and low bioavailability severely limit their biomedical applications. In the present study, BA and BE were encapsulated in our synthesized and amine-modified mesoporous silica nanoparticles (MSNs) (Nano-BA and Nano-BE, respectively), and their loading efficiencies and releasing profiles were investigated. Their cytotoxicity was examined on primary human gingival epithelial cells (hGECs), and the cellular uptake of Nano-BA or Nano-BE was visualized via a transmission electron microscope. Their anti-inflammatory effects were evaluated in IL-1β-treated hGECs using the cytokine array and enzyme-linked immunosorbent assay. The present study shows that the amine-modified MSNs could encapsulate BA and BE, and nano-encapsulation greatly enhances the drug delivery rate and prolongs the release of BA and BE up to 216 h. Moreover, both Nano-BA and Nano-BE could be internalized by hGECs and retained intracellularly in nanoparticle-free media for at least 24 h. Note that Nano-BE pre-treatment effectively down-regulates the IL-1β-induced expression of IL-6 and IL-8 in hGECs. In conclusion, nanoparticle-encapsulated BE exhibits notable anti-inflammatory effects through effective release and cellular internalization approaches. This study may facilitate the development of novel drug delivery systems for improving oral care. 10.1039/c7nr02546g
    Isoliquiritigenin alleviates P. gingivalis-LPS/ATP-induced pyroptosis by inhibiting NF-κB/ NLRP3/GSDMD signals in human gingival fibroblasts. Lv Xiaofang,Fan Chun,Jiang Zhongxin,Wang Wenxuan,Qiu Xu,Ji Qiuxia International immunopharmacology OBJECTIVE:To investigate whether pyroptosis is induced by Porphyromonas gingivalis-lipopolysaccharide (P. gingivalis-LPS)/ adenosine triphosphate (ATP) through NF-κB/NLRP3/GSDMD signaling in human gingival fibroblasts (HGFs) and whether isoliquiritigenin (ISL) alleviates pyroptosis by inhibition of NF-κB/NLRP3/GSDMD signals. DESIGN:Periodontitis was optimally simulated using a combination of P. gingivalis-LPS and ATP. The expression levels of genes and proteins of NF-κB, NLRP3 inflammasome, GSDMD, and IL-1β was characterized by qRT-PCR, western blotting and ELISA. The 2',7'‑dichlorodihydrofluorescein diacetate fluorescence probe was used to determine the intracellular ROS level. Hoechst 33342 and PI double staining, cytotoxicity assay, and caspase-1 activity assay were used to confirm the influence of ISL on pyroptosis in P. gingivalis-LPS/ATP-treated HGFs. RESULTS:P. gingivalis-LPS/ATP stimulation significantly promoted expression of NF-κB, the NLRP3 inflammasome, GSDMD, and IL-1β at gene and protein levels. The proportion of membrane-damaged cells, caspase-1 activity, and the release of lactate dehydrogenase (LDH) were also elevated. However, pretreatment with ISL observably suppressed these effects. CONCLUSIONS:P. gingivalis-LPS/ATP induced pyroptosis in HGFs by activating NF-κB/NLRP3/GSDMD signals and ISL attenuated P. gingivalis-LPS/ATP-induced pyroptosis by inhibiting these signals. This evidence may provide a new direction for the treatment of periodontitis. 10.1016/j.intimp.2021.108338
    Evaluation of the effect of oleuropein on alveolar bone loss, inflammation, and apoptosis in experimental periodontitis. Taskan Mehmet Murat,Balci Yuce Hatice,Karatas Ozkan,Gevrek Fikret,Toker Hülya Journal of periodontal research THE OBJECTIVE:The present study aimed to evaluate the effects of oleuropein on ligature-induced alveolar bone loss. In this respect, osteoblastic activity, osteoclastic activity, inflammatory markers, and apoptosis were evaluated. BACKGROUND:Oleuropein is a flavonoid, which has potent anti-inflammatory and bone-protective effects. METHODS:Thirty-two Wistar rats were divided into four experimental groups as following: control (C, n = 8) group; periodontitis (P, n = 8) group; periodontitis and low-dose oleuropein group (12 mg/kg/day oleuropein, LDO group, n = 8); and periodontitis and high-dose oleuropein group (24 mg/kg/day oleuropein, HDO group, n = 8). Periodontitis was induced via ligatures. Study period was 14 days, and animals were sacrificed at end of this period. Mandibles were examined via a stereomicroscope and underwent histological procedures. Osteoblast, tartrate-resistant acid phosphatase (TRAP)-positive osteoclast, and inflammatory cell counts were determined in hematoxylin-eosin stained sections. Inducible nitric oxide synthase (iNOS), bone morphogenetic protein-4, the cluster of differentiation (CD)-68, cysteine-aspartic proteases-3 (Caspase 3), and B-cell lymphoma-2 (Bcl-2) expressions were evaluated via immunohistochemistry. RESULTS:Periodontitis group had highest alveolar bone loss, and these levels significantly decreased in LDO and HDO groups. Both 12 and 24 mg/kg oleuropein groups significantly increased osteoblast cell counts and decreased TRAP-positive osteoclast and inflammatory cell counts. BMP-4 and bcl-2 expressions were elevated in oleuropein groups while caspase-3 expressions decreased. iNOS and CD68 were higher in periodontitis group compared to control group, but there was no significant difference between other groups. CONCLUSION:Oleuropein successfully decreased alveolar bone loss as a result of decreased osteoclastic activity, inflammation, and apoptosis and increased osteoblastic activity. 10.1111/jre.12662
    Quercetin protects oral mucosal keratinocytes against lipopolysaccharide-induced inflammatory toxicity by suppressing the AKT/AMPK/mTOR pathway. Immunopharmacology and immunotoxicology BACKGROUND:Cytokines can induce a chronic inflammatory response in the periodontium, leading to periodontitis. Quercetin, a naturally occuring flavonoid, has been shown to inhibit periodontitis, but how it works is poorly understood. In this study, we assessed the impact of quercetin on lipopolysaccharide (LPS)-induced inflammatory damage in oral mucosal keratinocytes (hOMK107) and explored its underlying mechanism. METHODS:The viability and apoptosis of hOMK107 cells were measured after exposure to LPS, followed or not by quercetin. The production of IL-1β, IL-6, IL-8, TNF-ɑ, iNOS, and COX-2 was quantified by enzyme-linked immunosorbent assay (ELISA), while levels of Akt, AMPK, and mTOR and their phosphorylation were detected semi-quantitatively by western blotting. RESULTS:Quercetin significantly improved cell viability and apoptosis by reversing LPS-induced upregulation of Bax and downregulation of Bcl-2 in hOMK107 cells. Quercetin decreased the production of IL-1β, IL-6, IL-8, TNF-ɑ, iNOS, and COX-2, as well as signal transduction the Akt/AMPK/mTOR pathway. Inhibitors of Akt, AMPK, and mTOR strengthened the anti-apoptotic effects of quercetin, while agonists of Akt, AMPK, or mTOR or Akt overexpression weakened the anti-apoptotic effects. CONCLUSION:These results indicate that quercetin may have a potential protective effect against the chronic inflammation-related periodontitis suppressing Akt/AMPK/mTOR pathway. 10.1080/08923973.2021.1948565
    Effects of rutin on the oxidative stress, proliferation and osteogenic differentiation of periodontal ligament stem cells in LPS-induced inflammatory environment and the underlying mechanism. Zhao Bin,Zhang Wenjing,Xiong Yixuan,Zhang Yunpeng,Zhang Dongjiao,Xu Xin Journal of molecular histology Periodontitis can cause damage to dental support tissue and affect the function of periodontal ligament cells. Rutin, a common flavonoid, plays a key role in anti-inflammatory responses, tissue repair and bone development. The purpose of this study was to investigate the effects of rutin on the oxidative stress, proliferation, and osteogenic differentiation of human periodontal ligament stem cells (PDLSCs) in an inflammatory environment and the underlying mechanism. Lipopolysaccharide (LPS) was used to stimulate PDLSCs to mimic an inflammatory environment model. Reactive oxygen species (ROS) levels were detected by the dichlorodihydrofluorescein diacetate (DCFH-DA) probe and the oxidative stress factors were tested by an oxidative stress factor detection kit. Moreover, the proliferation of PDLSCs was evaluated by cell counting kit-8 (CCK-8) assay. In addition, the osteogenic differentiation of PDLSCs was determined by alkaline phosphatase (ALP) staining, ALP activity test, alizarin red staining, and alizarin red semi-quantitative analysis. Furthermore, the protein levels of AKT and p-AKT were detected by Western blot. The results showed that rutin inhibited the release of ROS and increased the secretion of oxidative stress factors [superoxide dismutase (SOD) and glutathione (GSH)] and promoted the proliferation of PDLSCs in an inflammatory environment. Moreover, rutin upregulated ALP activity and enhanced the number of mineralized nodules. Conversely, the use of LY294002 (an inhibitor of PI3K) blocked the activation of the PI3K/AKT signaling pathway and prevented the beneficial effects of rutin. In conclusion, rutin promoted the antioxidative stress ability, proliferation and osteogenic differentiation of PDLSCs through PI3K/AKT signaling pathway in LPS-induced inflammatory environment. 10.1007/s10735-020-09866-9
    Assessing the effect and related mechanism of naringenin on the proliferation, osteogenic differentiation and endothelial differentiation of human periodontal ligament stem cells. Zhang Li,He Haiyan,Zhang Min,Wu Yujie,Xu Xiaomei,Yang Maohua,Mei Li Biochemical and biophysical research communications Naringenin (NAR) is a natural flavonoid which exerts extensive biological activity, including anti-oxidation, anti-inflammation, anti-cancer, immune regulation and so on. However, the effect and mechanism of NAR in the alveolar bone regeneration are still unclear, which limits its clinical use. Hence, we investigated the effects of NAR in the proliferation, osteogenic and endothelial differentiation of human periodontal ligament stem cells (hPDLSCs) and explore the possible mechanism. The results showed that the proper concentrations (100 nM-10 μM) of NAR can promote the proliferation rate, osteogenic and endothelial differentiation of hPDLSCs. And the 1 μM NAR had the best proliferation promoting effect, while the 10 μM NAR had the best ability of promoting osteogenic and endothelial differentiation. NAR also promoted the mRNA expression of SDF-1 in a concentration dependent manner in PDLSCs. After adding the selective CXCR4 antagonist AMD3100, the osteogenic effect of NAR on PDLSCs is slightly enhanced, while the endothelial differentiation effect of NAR on hPDLSCs is attenuated. In summary, these results indicated that NAR promoted the proliferation of hPDLSCs, and promoted endothelial differentiation of hPDLSCs via SDF-1 to activate SDF-1/CXCR4 signaling pathway. However, the mechanism of which SDF-1 related signaling pathway is activated by NAR to enhance the osteogenic differentiation of hPDLSCs still needs to be investigated. 10.1016/j.bbrc.2020.11.081
    Kaempferol promotes proliferation and osteogenic differentiation of periodontal ligament stem cells via Wnt/β-catenin signaling pathway. Nie Fujiao,Zhang Wenjuan,Cui Qun,Fu Yajing,Li Hongkun,Zhang Jun Life sciences AIMS:Kaempferol, a type of flavonoid, is widely present in fruits, vegetables and medicinal herbs. This study was designed to investigate the effects of kaempferol on proliferation and osteogenesis of periodontal ligament stem cells (PDLSCs) and to identify the related pathway. MATERIALS AND METHODS:PDLSCs were isolated from extracted premolars and cultured in vitro. Cell-counting kit-8 (CCK-8) and colony formation assays were performed to determine the effect of kaempferol, at various concentrations, on the proliferation of PDLSCs. Alkaline phosphatase (ALP) activity was analyzed both quantitatively and qualitatively, and extracellular matrix mineralization was examined by alizarin red-S staining. In addition, the mRNA and protein expression levels of ALP, RUNX Family Transcription Factor 2 (RUNX2), Sp7 Transcription Factor (SP7; Osterix), Bone Gamma-Carboxyglutamate Protein (BGLAP; osteocalcin) and catenin beta 1 (CTNNB1; β-catenin) were monitored by qPCR and Western blot analysis. Additionally, the tankyrase inhibitor, XAV939, was used to determine the role of the Wnt/β-catenin pathway. KEY FINDINGS:The results illustrated that 10 M kaempferol markedly promoted the proliferation, ALP activity and mineral deposition of PDLSCs (P < 0.05). The expression levels of ALP, RUNX2, SP7, BGLAP and β-catenin were all upregulated (P < 0.05). After blocking the Wnt/β-catenin pathway with XAV939, the effects of kaempferol were apparently reversed. SIGNIFICANCE:kaempferol enhanced proliferation and osteogenesis of PDLSCs by activating the Wnt/β-catenin signaling, which suggests the potential application of kaempferol for periodontal tissue regeneration. 10.1016/j.lfs.2020.118143
    Inhibitory Effects of Standardized Boesenbergia pandurata Extract and Its Active Compound Panduratin A on Lipopolysaccharide-Induced Periodontal Inflammation and Alveolar Bone Loss in Rats. Kim Haebom,Kim Changhee,Kook Kyo Eun,Yanti ,Choi Seungmok,Kang Wonku,Hwang Jae-Kwan Journal of medicinal food Periodontitis, an inflammatory disease of the gingival tissue, triggered by microbial-derived elements, such as lipopolysaccharide (LPS), collapses the periodontal tissues and resorbs the alveolar bone. This study evaluated the inhibitory effects of standardized Boesenbergia pandurata extract (BPE) and panduratin A (PAN) on periodontitis-induced inflammation and alveolar bone loss. Sprague-Dawley rats with LPS-induced periodontitis were orally administered BPE (50 and 200 mg/kg/day) and PAN (20 mg/kg/day) for 8 days. Histological analysis revealed that BPE- and PAN-administered groups showed decreased cell infiltration and alveolar bone resorption. Furthermore, the BPE and PAN significantly alleviated the mRNA and protein expression levels of nuclear factor kappa B (NF-κB), interleukin-1β, matrix metalloproteinase (MMP)-2, and MMP-8. BPE and PAN also inhibited the expression of nuclear factor of activated T cells, cytoplasmic 1, c-Fos, and ostoclastogenesis-related enzymes, including cathepsin K and tartrate-resistant acid phosphatase (ALP). BPE and PAN not only upregulated the osteoblastogenesis-associated markers, such as collagen type I (COL1A1) and ALP, but also increased the ratio of osteoprotegerin to receptor activator of NF-κB ligand. Collectively, BPE and PAN efficiently prevent destruction of periodontal tissues and stimulating the loss of alveolar bone tissues, strongly indicative of their potential as natural antiperiodontitis agents. 10.1089/jmf.2017.4155
    Quercetin Decreased Alveolar Bone Loss and Apoptosis in Experimentally Induced Periodontitis Model in Wistar Rats. Taskan Mehmet Murat,Gevrek Fikret Anti-inflammatory & anti-allergy agents in medicinal chemistry BACKGROUND:Quercetin is a flavonoid which has potent anti-inflammatory, antibacterial, and antioxidant effect. Purpose of this study was to evaluate effects of quercetin on alveolar bone loss and histopathological changes in ligature-induced periodontitis in rats. METHODS:Wistar rats were divided into four experimental groups: non-ligated control (C, n=8) group; periodontitis (P, n=8) group; ligature and low dose quercetin group (75 mg/kg/day quercetin, Q75 group, n=8); ligature and high dose quercetin group (150 mg/kg/day quercetin, Q150 group, n=8). Silk ligatures were placed at gingival margin of lower first molars of mandibular right quadrant. Study duration was 15 days, and animals were sacrificed end of this period. Changes in alveolar bone levels were clinically measured and tissues were immunohistochemically examined, matrix metalloproteinase 8 (MMP 8), inducible nitric oxide synthase (iNOS), tissue inhibitor of metalloproteinase 1 (TIMP 1), Cysteine-aspartic proteases 3 (Caspase 3), and tartrate-resistant acid phosphatase (TRAP) positive osteoclast cells, osteoblast, and neutrophil counts were also determined. RESULTS AND DISCUSSION:Alveolar bone loss was highest in P group, and differences among P, Q75, and Q150 groups were significant. Both doses of quercetin decreased TRAP+ osteoclast cells and increased osteoblast cells. Inflammation in P group was also higher than those of C, Q75, and Q150 groups indicating anti-inflammatory effect of quercetin. iNOS, MMP-8, and caspase-3 levels were highest, and TIMP-1 expression was lowest in P group; differences were statistically significant. CONCLUSION:Within limits of this study, it can be suggested that quercetin administration may reduce alveolar bone loss by increasing osteoblastic activity, decreasing osteoclastic activity, apoptosis, and inflammation in an experimental model of periodontitis. 10.2174/1871523019666200124114503
    The effects of taxifolin on alveolar bone in experimental periodontitis in rats. Lektemur Alpan Aysan,Kızıldağ Alper,Özdede Melih,Karakan Nebi Cansın,Özmen Özlem Archives of oral biology OBJECTIVE:This study aimed to evaluate the effect of taxifolin, a powerful antioxidant, on the progression of periodontitis by immunohistochemical and cone-beam computed tomography (CBCT) examination. DESIGN:This study was performed with 32 rats in four experimental groups: a non-ligated group (Control, n = 8), periodontitis group (Perio, n = 8), periodontitis with 1 mg/kg/day taxifolin group (Taxi-1, n = 8), and periodontitis with 10 mg/kg/day taxifolin group (Taxi-10, n = 8). A ligature-induced experimental periodontitis design was used. All rats were sacrificed at 30 days. Alveolar bone loss was determined by CBCT. Hematoxylin-eosin stained slides were examined. The expression levels of bone morphogenetic protein 2 (BMP-2), osteocalcin (OCN), alkaline phosphatase (ALP), collagen type I (Col 1), Bcl-2, Bax, and receptor activator of NF-κB ligand (RANKL) were determined immunohistochemically. RESULTS:Both doses of taxifolin showed a decrease in alveolar bone loss. The inflammatory reaction was higher in the Perio group and lower in the taxifolin groups. BMP-2, OCN, ALP, and Col 1 expression were dose-dependently elevated in the taxifolin groups. RANKL immunoexpression decreased with both doses of taxifolin. Bcl-2 expression increased and Bax expression decreased in the taxifolin groups. CONCLUSION:Taxifolin successfully reduced apoptosis and improved bone formation in alveolar bone in this experimental periodontitis model. 10.1016/j.archoralbio.2020.104823
    Ipriflavone promotes proliferation and osteogenic differentiation of periodontal ligament cells by activating GPR30/PI3K/AKT signaling pathway. Han Yuanyuan,Wang Xuxia,Ma Dan,Wu Xiaoxiao,Yang Panpan,Zhang Jun Drug design, development and therapy OBJECTIVES:This study was performed to investigate the effects and mechanism of ipriflavone (IP) on the proliferation and osteoblastic differentiation of periodontal ligament cells in vitro and periodontal tissue remodeling following orthodontic tooth movement (OTM) in vivo. MATERIALS AND METHODS:Human periodontal ligament cells (hPDLCs) were cultured in vitro and cell counting kit-8, alkaline phosphatase (ALP) activity assay, plate clone formation assay, and alizarin red staining were used to test proliferation and osteogenic differentiation of hPDLCs. What is more, the expression of ALP, Runx2, and GPR30 was examined by real-time polymerase chain reaction and Western blot. To find out if PI3K/AKT signaling pathway was involved in the process, AKT and p-AKT were examined by Western blot. LY294002 (PI3K signaling pathway inhibitor) and small interfering RNA targeting GPR30 mRNA (siGPR30) were used to verify the function of GPR30-mediated PI3K/AKT pathway in this process. Twenty-four male Wistar rats were randomized into 2 groups, the control group with force application and the IP group with force application plus IP. Morphological changes in the periodontal tissue between roots of teeth were investigated using hematoxylin and eosin (HE) staining and bone morphogenetic protein-2 was detected to assess bone remodeling by immunohistochemical staining. RESULTS:In vitro, 10 M IP was selected significantly promoting proliferation, ALP activity, colony forming efficiency, and mineral deposition (<0.05) on hPDLCs. Gene expressions of , , , and were all upregulated than the control group (<0.05). According to the mechanism, promotion of ALP and Runx2 interdicted by LY294002 and siGPR30 reduced the activation of PI3K/AKT signaling pathway. In addition, HE staining and immunohistochemical staining results showed that the IP group had more new bone formation in the periodontal tissue compared to the control group in vivo. CONCLUSION:IP can promote the expression of ALP and Runx2 which was probably related to the GPR30-mediated PI3K/AKT signaling pathway. Moreover, IP coordination seemed to have the potential to prevent relapsing following OTM. 10.2147/DDDT.S148457
    Epigallocatechin gallate affects the proliferation of human alveolar osteoblasts and periodontal ligament cells, as well as promoting cell differentiation by regulating PI3K/Akt signaling pathway. Ding Cheng,Fu Shulei,Chen Xing,Chen Chongchong,Wang Huiming,Zhong Liangjun Odontology Human periodontal ligament cells (hPDLCs) and human alveolar osteoblasts (hAOBs) play pivotal roles in periodontium. The regulatory effects of epigallocatechin gallate (EGCG) on hPDLCs and hAOBs remained unclear. This study probed into the functions of EGCG treating periodontal diseases. Cultured hAOBs and hPDLCs were passaged and observed by microscopic examination, and alkaline phosphatase (ALP) and immumohistochemical staining were performed for verification. hAOBs and hPDLCs were treated with EGCG and LY294002 + EGCG, then the proliferation of the two cells was assayed by MTT. Mineralization of the treated hAOBs and hPDLCs was detected by ALP activity experiment and Alizarin Red S staining kit. Quantitative real-time polymerase chain reaction (qRT-PCR) and Western blotting were performed for the detection of the expressions of differentiation-related mRNAs and PI3K/Akt signaling pathway-related proteins in the two cells. The third passage of hAOBs mainly showed triangle shape and were positive by ALP staining. hPDLCs in passage 3 adhered to the wall in spiral or radial pattern with positively stained vimentin and negatively stained keratin. Cell proliferation and ALP activity of the hAOBs and hPDLCs were increased by EGCG treatment. The mineralized nodules and expressions of differentiation-related mRNAs, the phosphorylation of PI3K and Akt of the hAOBs and hPDLCs were promoted by EGCG treatment, while the effects of LY294002 treatment were opposite to EGCG treatment. Epigallocatechin gallate affected the proliferation and differentiation of hAOBs and hPDLCs through regulating PI3K/Akt signaling pathway. 10.1007/s10266-021-00597-1
    The effect of luteolin in prevention of periodontal disease in Wistar rats. Balci Yuce Hatice,Toker Hulya,Yildirim Ali,Tekin Mehmet Bugrul,Gevrek Fikret,Altunbas Nilufer Journal of periodontology BACKGROUND:Periodontal disease is the chronic infectious disease of the periodontium. Because of irreversibility, prevention of disease is one of the most important goals of periodontal treatment. The aim of this study was to evaluate the effect of luteolin, a powerful anti-inflammatory agent, on the prevention of experimental periodontitis by determining morphological and histological tissue alterations. METHODS:This study consisted of 28 rats and four experimental groups: healthy control group (C, n = 6); periodontitis group (P, n = 6); periodontitis and 50 mg/kg luteolin administered group (L-50, n = 8); and periodontitis and 100 mg/kg luteolin administered group (L-100, n = 8). Experimental periodontitis was induced via ligature method around lower right first molar teeth. All rats were euthanized 11 days after. The severity of periodontal destruction was determined by measuring alveolar bone loss under a stereomicroscope. Osteoblast and inflammatory cell counts were counted on hematoxylin-eosin-stained slides and osteoclasts were counted on tartrate-resistant acid phosphatase-stained slides. The levels of inducible nitric oxide synthase (iNOS), bone morphogenetic protein (BMP)-2, matrix metalloproteinase (MMP)-8, tissue inhibitor of MMP (TIMP)-1, receptor activator of nuclear factor κB ligand (RANKL), and osteoprotegerin (OPG) were determined by immunohistochemistry. RESULTS:The highest alveolar bone loss was observed in the periodontitis group and the luteolin administration decreased bone loss in both groups. Osteoblast cell number was higher and osteoclast and inflammatory cell numbers were lower in the P group compared to C, L-50, and L-100 groups. Luteolin, dose-dependently increased osteoblast cell counts. Luteolin attenuated periodontal inflammation in both L-50 and L-100 groups. Like osteoblast cell numbers, BMP-2 expressions were also elevated in luteolin groups. Both doses of luteolin significantly increased TIMP-1 and BMP-2 expressions and decreased MMP-8 levels. iNOS expressions increased in P group and L-100 significantly decreased iNOS levels. RANKL increased and OPG decreased in P group and 100 mg/kg luteolin increased OPG and decreased RANKL levels significantly. CONCLUSIONS:Within the limits of present experimental study, luteolin successfully improved periodontal health in a ligature-induced experimental periodontitis model in Wistar rats. The decrease in inflammation, osteoclastic and collagenase activity and increase in osteoblastic activity are possibly involved in this process. 10.1002/JPER.18-0584
    Characteristics and biologic effects of thermosensitive quercetin-chitosan/collagen hydrogel on human periodontal ligament stem cells. Arpornmaeklong Premjit,Sareethammanuwat Maytha,Apinyauppatham Komsan,Boonyuen Supakorn Journal of biomedical materials research. Part B, Applied biomaterials Thermosensitive hydrogels could function as scaffolds and delivery vehicle of natural flavonoids. The current study aimed to investigate effects of chitosan/collagen ratios on properties of thermosensitive beta-glycerophosphate (bGP) chitosan/collagen hydrogels as delivery vehicle of quercetin and then examined effects of quercetin-hydrogels on growth and cell viability of human periodontal ligament stem cells (hPDLSCs). Microstructure and physical, mechanical and antioxidant properties and quercetin release profiles of the hydrogels were investigated. Fourier transform infrared spectroscopy and X-ray powder diffraction analyses were performed to examine gelation process of the hydrogels. Antioxidant assays were conducted to measure antioxidant capacity of quercetin-hydrogels. It was found that bGP-chitosan/collagen hydrogels exhibited porous structures with interconnected pore architecture and could sustain quercetin release. Chitosan content improved well defined porous structure, increased porosity of the hydrogels and decreased releasing rate of quercetin from the hydrogels. The quercetin-bGP-2:1 (wt/wt) chitosan/collagen hydrogels exhibited antioxidant capacity and were able to promote growth of hPDLSCs in a dose dependent manner. In conclusion, the thermosensitive quercetin-bGP-2:1 (wt/wt) chitosan/collagen hydrogel demonstrated optimal properties of scaffolds for bone tissue engineering and sustained release of natural flavonoids. Incorporating quercetin in the chitosan/collagen hydrogel enhanced bioactive microenvironment that supported stem cell encapsulation. 10.1002/jbm.b.34823
    Fisetin attenuates periodontitis through FGFR1/TLR4/NLRP3 inflammasome pathway. Huang Xin,Shen Hong,Liu Yiran,Qiu Sainan,Guo Yan International immunopharmacology The purpose of the present study was to investigate the pharmacological effect of Fisetin on experimental periodontitis in rats and explore its potential mechanism. The ligature/LPS method was used to induce periodontitis in rats. LPS was employed to cause inflammation in Human gingival fibroblasts (HGF). The transfections with FGFR1 SiRNA, NLRP3 SiRNA and the selective TLR4 inhibitor TAK242 were used to investigate the mechanism of Fisetin-mediated inflammatory reaction in LPS-induced HGF. As a result, Fisetin reduced the alveolar bone gap, reversed histopathological lesion and inhibited serum inflammatory cytokine concentration in periodontitis rats. Fisetin decreased the inflammatory cytokine contents in the supernatant of LPS-induced HGF. The inhibitory effect of Fisetin might be attributed to FGFR1/TLR4/NLRP3 inflammasome pathway both in vivo and in vitro. The suppressions of FGFR1, TLR4 and NLRP3 proved that FGFR1/TLR4/NLRP3 signaling was involved in the Fisetin-mediated inflammatory response. Fisetin also inhibited NLRP3 priming. The data demonstrated that Fisetin attenuated periodontitis by inhibiting inflammatory reaction via FGFR1/TLR4/NLRP3 inflammasome pathway. 10.1016/j.intimp.2021.107505
    Local icariin application enhanced periodontal tissue regeneration and relieved local inflammation in a minipig model of periodontitis. Zhang Xiuli,Han Nannan,Li Guoqing,Yang Haoqing,Cao Yangyang,Fan Zhipeng,Zhang Fengqiu International journal of oral science Periodontitis is an inflammatory autoimmune disease. Treatment should alleviate inflammation, regulate the immune reaction and promote periodontal tissue regeneration. Icariin is the main active ingredient of Epimedii Folium, and it is a promising compound for the enhancement of mesenchymal stem cell function, promotion of bone formation, inhibition of bone resorption, alleviation of inflammation and regulation of immunity. The study investigated the effect of icariin on periodontal tissue regeneration in a minipig model of periodontitis. The minipig model of periodontitis was established. Icariin was injected locally. The periodontal clinical assessment index, a computed tomography (CT) scan, histopathology and enzyme-linked immune sorbent assay (ELISA) were used to evaluate the effects of icariin. Quantitative analysis results 12 weeks post-injection demonstrated that probing depth, gingival recession, attachment loss and alveolar bone regeneration values were (3.72 ± 1.18) mm vs. (6.56 ± 1.47) mm, (1.67 ± 0.59) mm vs. (2.38 ± 0.61) mm, (5.56 ± 1.29) mm vs. (8.61 ± 1.72) mm, and (25.65 ± 5.13) mm vs. (9.48 ± 1.78) mm in the icariin group and 0.9% NaCl group, respectively. The clinical assessment, CT scan, and histopathology results demonstrated significant enhancement of periodontal tissue regeneration in the icariin group compared to the 0.9% NaCl group. The ELISA results suggested that the concentration of interleukin-1 beta (IL-1β) in the icariin group was downregulated compared to the 0.9% NaCl group, which indicates that local injection of icariin relieved local inflammation in a minipig model of periodontitis. Local injection of icariin promoted periodontal tissue regeneration and exerted anti-inflammatory and immunomodulatory function. These results support the application of icariin for the clinical treatment of periodontitis. 10.1038/s41368-018-0020-3
    Quercetin Prevents Oxidative Stress-Induced Injury of Periodontal Ligament Cells and Alveolar Bone Loss in Periodontitis. Drug design, development and therapy PURPOSE:Emerging evidence has indicated that oxidative stress (OS) contributes to periodontitis. Periodontal ligament cells (PDLCs) are important for the regeneration of periodontal tissue. Quercetin, which is extracted from fruits and vegetables, has strong antioxidant capabilities. However, whether and how quercetin affects oxidative damage in PDLCs during periodontitis remains unknown. The aim of this study was to assess the effects of quercetin on oxidative damage in PDLCs and alveolar bone loss in periodontitis and underlying mechanisms. MATERIALS AND METHODS:The tissue block culture method was used to extract human PDLCs (hPDLCs). First, a cell counting kit 8 (CCK-8) assay was used to identify the optimal concentrations of hydrogen peroxide (HO) and quercetin. Subsequently, a 2,7-dichlorodihydrofluorescein diacetate (DCFH-DA) probe, RT-qPCR, Western blotting and other methods were used to explore the effects of quercetin on OS in hPDLCs and the underlying mechanism. Finally, quercetin was administered to mice with periodontitis through gavage, and the effect of quercetin on the level of OS and alveolar bone resorption in these mice was observed by immunofluorescence, microcomputed tomography (micro-CT), hematoxylin and eosin staining (H&E) staining and so on. RESULTS:Quercetin at 5 μM strongly activated NF-E2-related factor 2 (NRF2) signaling, alleviated oxidative damage and enhanced the antioxidant capacity of hPDLCs. In addition, quercetin reduced cellular senescence and protected the osteogenic ability of hPDLCs. Finally, quercetin activated NRF2 signaling in the periodontal ligaments, reduced the OS level of mice with periodontitis, and slowed the absorption of alveolar bone in vivo. CONCLUSION:Quercetin can increase the antioxidant capacity of PDLCs and reduce OS damage by activating the NRF2 signaling pathway, which alleviates alveolar bone loss in periodontitis. 10.2147/DDDT.S315249
    Quercetin-Loaded Ceria Nanocomposite Potentiate Dual-Directional Immunoregulation via Macrophage Polarization against Periodontal Inflammation. Wang Yu,Li Chunyan,Wan Yao,Qi Manlin,Chen Qiuhan,Sun Yue,Sun Xiaolin,Fang Jiao,Fu Li,Xu Lin,Dong Biao,Wang Lin Small (Weinheim an der Bergstrasse, Germany) Macrophage polarization toward M1 phenotype (pro-inflammation) is closely associated with the destructive phase of periodontal inflammation. Nanoceria is verified to inhibit M1 polarization of macrophages by the favorable ability of reactive oxygen species (ROS) scavenging. However, the function of nanoceria on macrophage polarization toward M2 phenotype (anti-inflammation) in reparative phase of periodontal inflammation is quite limited. In this work, by introducing an antioxidant drug quercetin onto nano-octahedral ceria, synergistic and intense regulation of host immunity against periodontal disease is realized. Such nanocomposite can control the phenotypic switch of macrophages by not only inhibition of M1 polarization for suppressing the damage in the destructive phase but also promotion of M2 polarization for regenerating the surrounding tissues in reparative phase of periodontal disease. As-prepared nanocomposite can effectively increase the M2/M1 ratio of macrophage polarization in inflammatory cellular models by lipopolysaccharide stimulation. More importantly, the nanocomposite also exerts an improved therapeutic potential against local inflammation by significant downregulation of pro-inflammatory cytokines and upregulation of anti-inflammatory cytokines in an animal model with periodontal inflammation. Therefore, this newly developed nanomedicine is efficient in ROS scavenging and driving pro-inflammatory macrophages to the anti-inflammatory phenotype to eliminate inflammation, thereby providing a promising candidate for treating periodontal inflammation. 10.1002/smll.202101505
    Icariin induces the growth, migration and osteoblastic differentiation of human periodontal ligament fibroblasts by inhibiting Toll-like receptor 4 and NF-κB p65 phosphorylation. Liu Hai-Jiang,Liu Xue-Yang,Jing De-Bao Molecular medicine reports The proliferation, migration and differentiation capacities of human periodontal ligament fibroblasts (HPDLCs) are important for the treatment of periodontal diseases. The aim of the present study was to investigate whether icariin could promote these abilities in HPDLCs, and explore the cellular mechanisms therein. The results indicated that icarrin markedly blocked apoptosis, and increased the viability and migration of HPDLCs, particularly at the concentrations of 20 and 50 µM. In addition, icariin significantly promoted HPDLCs to synthesize extracellular matrix, which was reflected by the decreased expression of matrix matalloproteinase-1 and increased expression of tissue inhibitor of metalloproteinase-1. Furthermore, the levels of bone morphogenetic protein 2, collagen I, osteoprotegerin and alkaline phosphatase were markedly elevated by icariin, indicating that icariin was able to promote the osteogenic differentiation capability of HPDLCs. Icariin also inactivated the Toll-like receptor 4 (TLR)-4/nuclear factor (NF)-κB signaling pathway by suppressing the expression levels of TLR-4 and phosphorylated p65, and by blocking p65 nuclear translocation. These results suggested that icarrin increased the survival, migration and osteoblastic differentiation of HPDLCs by inhibiting the TLR-4/NF-κB signaling pathway. 10.3892/mmr.2018.9302
    The effect of ethanolic extract of Brazilian green propolis and artepillin C on aFGF-1, Eselectin, and CD40L secreted by human gingival fibroblasts. Central-European journal of immunology INTRODUCTION:Periodontal diseases are among the most common diseases of the oral cavity in the worldwide population. The prevention of gingivitis and periodontitis is based on the removal of bacterial plaque from the teeth with use of toothpaste containing active substances. Noteworthy is the ethanolic extract of Brazilian green propolis (EEP-B), which, due to the high content of artepillin C, has anti-inflammatory, antibacterial, or immunostimulatory effects. Little is known about interactions between EEP-B and gingival fibroblasts within the oral cavity. The purpose of the article is to determine the role of acidic fibroblast growth factor (aFGF-1), E-selectin, and ligand of CD40 (CD40L) secreted by human gingival fibroblasts (HGF-1) in the gingiva. MATERIAL AND METHODS:We performed our experiments on gingival fibroblasts (HGF-1), which are an ideal in vitro model for studying the processes taking place within the gingiva. We incubated cells with EEP-B or artepillin C at the concentrations of 1 µg/ml and 10 µg/ml. The aFGF-1, E-selectin, and CD40L were detected using the Bio-Plex Magnetic Luminex Assay and the Bio-Plex 200 System. RESULTS:Ethanolic extract of Brazilian green propolis and artepillin C increased the levels of aFGF-1 secreted by HGF-1. Moreover, EEP-B decreased the levels of E-selectin in both tested concentrations, which was not proved for artepillin C. No changes in the concentration of CD40L released by HGF-1 were observed. CONCLUSIONS:The obtained results may suggest that EEP-B, due to the mixture of various compounds including flavonoids, accelerates the wound healing effects and has anti-inflammatory activity. 10.5114/ceji.2021.111215
    Treatment with Luteolin Improves Lipopolysaccharide-Induced Periodontal Diseases in Rats. Casili Giovanna,Ardizzone Alessio,Lanza Marika,Gugliandolo Enrico,Portelli Marco,Militi Angela,Cuzzocrea Salvatore,Esposito Emanuela,Paterniti Irene Biomedicines Periodontitis is a dental disease that produces the progressive destruction of the bone surrounding the tooth. Especially, lipopolysaccharide (LPS) is involved in the deterioration of the alveolar bone, inducing the release of pro-inflammatory mediators, which cause periodontal tissue inflammation. Luteolin (Lut), a molecule of natural origin present in a large variety of fruits and vegetables, possess beneficial properties for human health. On this basis, we investigated the anti-inflammatory properties of Lut in a model of periodontitis induced by LPS in rats. Animal model predicted a single intragingival injection of LPS (10 μg/μL) derived from . Lut administration, was performed daily at different doses (10, 30, and 100 mg/kg, orally), starting from 1 h after the injection of LPS. After 14 days, the animals were sacrificed, and their gums were processed for biochemical analysis and histological examinations. Results showed that Lut (30 and 100 mg/kg) was equally able to reduce alveolar bone loss, tissue damage, and neutrophilic infiltration. Moreover, Lut treatment reduced the concentration of collagen fibers, mast cells degranulation, and NF-κB activation, as well as the presence of pro-inflammatory enzymes and cytokines. Therefore, Lut implementation could represent valid support in the pharmacological strategy for periodontitis, thus improving the well-being of the oral cavity. 10.3390/biomedicines8100442
    Tea polyphenols inhibit the activation of NF-κB and the secretion of cytokines and matrix metalloproteinases by macrophages stimulated with Fusobacterium nucleatum. Lagha Amel Ben,Grenier Daniel Scientific reports Fusobacterium nucleatum has been associated with both periodontal disease and inflammatory bowel disease. This Gram-negative bacterium possesses a high inflammatory potential that may contribute to the disease process. We hypothesized that green and black tea polyphenols attenuate the inflammatory response of monocytes/macrophages mediated by F. nucleatum. We first showed that the tea extracts, EGCG and theaflavins reduce the NF-κB activation induced by F. nucleatum in monocytes. Since NF-κB is a key regulator of genes coding for inflammatory mediators, we tested the effects of tea polyphenols on secretion of IL-1β, IL-6, TNF-α, and CXCL8 by macrophages. A pre-treatment of macrophages with the tea extracts, EGCG, or theaflavins prior to a stimulation with F. nucleatum significantly inhibited the secretion of all four cytokines and reduced the secretion of MMP-3 and MMP-9, two tissue destructive enzymes. TREM-1 expressed by macrophages is a cell-surface receptor involved in the propagation of the inflammatory response to bacterial challenges. Interestingly, tea polyphenols inhibited the secretion/shedding of soluble TREM-1 induced by a stimulation of macrophages with F. nucleatum. The anti-inflammatory properties of tea polyphenols identified in the present study suggested that they may be promising agents for the prevention and/or treatment of periodontal disease and inflammatory bowel disease. 10.1038/srep34520
    [Effect of resveratrol on expression of TLR4 and inflammatory factors in gingival epithelial cells under high glucose environment]. Lv Jia-Shu,Jiang Xue-Wei,Zhang Yan,Zhen Lei Shanghai kou qiang yi xue = Shanghai journal of stomatology PURPOSE:Through a study of the molecular mechanism of the effect of resveratrol(RSV) on expression of TLR4 and inflammatory factors in gingival epithelial cells under high glucose environment, the therapeutic effect and molecular mechanism of resveratrol on periodontitis in patients with diabetes mellitus was investigated. METHODS:Gingival epithelial cells were cultured in vitro; according to the way of action, the cultured cells were divided into control group, high glucose group(HG) and HG+RSV group. The mRNA expression of TLR4 was detected by PCR; The third generation of gingival epithelial cells were pre-treated with or without RSV for 24 h under high glucose conditions, and subsequently treated with LPS at 100 ng/mL for 2 h. ELISA was used to detect the secretion of IL-1 beta, IL-6, IL-8 and TNF- alpha; the activation of TLR4 downstream signaling molecules NF-κB p65, p38 MAPK, and STAT3 was determined by Western blot. SPSS17.0 software package was used for statistical analysis. RESULTS:RSV could reverse the increase of TLR4 level in gingival epithelial cells in high glucose medium.LPS markedly increased the expression and secretion of IL-1β, IL-6, IL-8, and TNF-α in GECs cultured in high glucose medium, which was partly blocked in the presence of RSV. Furthermore, Western blot results showed that RSV significantly suppressed the phosphorylation of TLR4 downstream factors NF-κB p65, p38MAPK, and STAT3. CONCLUSIONS:RSV reduces inflammatory cytokine secretion in gingival epithelial cells, through negative regulation of TLR4 signaling pathway.
    Regulation of matrix metalloproteinase secretion by green tea catechins in a three-dimensional co-culture model of macrophages and gingival fibroblasts. Morin Marie-Pierre,Grenier Daniel Archives of oral biology OBJECTIVES:Elevated levels of matrix metalloproteinases (MMPs) have been associated with the active phases of tissue and bone destruction in periodontitis, an inflammatory disease characterized by a significant breakdown of tooth support. In the present study, we used a three-dimensional (3D) co-culture model of macrophages and gingival fibroblasts to investigate the ability of a green tea extract and its major constituent epigallocatechin-3-gallate (EGCG) to regulate the secretion of MMP-3, -8, and -9. METHODS:The 3D co-culture model was composed of gingival fibroblasts embedded in a type I collagen matrix overlaid with macrophages. Two arbitrary ratios were tested. The ratio composed of 1 macrophage to 10 fibroblasts was used to mimic a slightly inflamed periodontal site while the ratio composed of 10 macrophages to 1 fibroblast was used to mimic a severely inflamed periodontal site. The 3D co-culture model was pre-treated for 2h with either the green tea extract or EGCG. It was then stimulated with Aggregatibacter actinomycetemcomitans lipopolysaccharide (LPS). The model was also first stimulated with LPS for 2h and then incubated with the green tea extract or EGCG. The concentrations of secreted MMP-3, -8, and -9 were quantified by enzyme-linked immunoassays. RESULTS:When the 3D co-culture model was stimulated with A. actinomycetemcomitans LPS, the 10:1 ratio of macrophages to gingival fibroblasts was associated with a highest secretion of MMP-3 and -9 and, to a lesser extent, MMP-8, than the 1:10 ratio. Non-cytotoxic concentrations of the green tea extract or EGCG reduced the basal secretion levels of all three MMPs. A 2-h treatment with the green tea extract or EGCG prior to the stimulation with LPS resulted in a dose-dependent decrease in MMP secretion, with MMP-9 showing the most significant decrease. A decrease in MMP secretion was also observed when the green tea extract or EGCG was added following a 2-h stimulation with LPS. CONCLUSIONS:Our results suggested that green tea catechins, and more specifically EGCG, offer promising prospects for the development of a novel adjunctive treatment for periodontitis because of their ability to decrease the secretion of MMPs, which are important tissue-destructive enzymes produced by mucosal and immune cells. 10.1016/j.archoralbio.2016.10.035
    Cytotoxicity and potential anti-inflammatory activity of velutin on RAW 264.7 cell line differentiation: Implications in periodontal bone loss. Brito Carlos,Stavroullakis Alexander,Oliveira Tatiane,Prakki Anuradha Archives of oral biology OBJECTIVES:Hypoxia-inducible factor-1α (HIF-1α) has been implicated in periodontal tissue inflammation and possibly in osteoclast differentiation, while polyphenols are known to be anti-inflammatory natural compounds that are capable of regulating the NF-κB protein complex pathway. The objective of this study was to investigate cytotoxicity and HIF-1α expression through the NF-κB pathway by polyphenol velutin (Euterpe oleracea Mart.), found in the pulp of acai fruit, during inflammatory RAW 264.7 differentiation. DESIGN:RAW 264.7 mouse monocyte macrophage cells were stimulated with RANKL (30ng/mL) and Porphyromonas gingivalis lipopolysaccharide (1μg/mL). Cells were treated with various concentrations of velutin (0.5-2μM) to check for viability, morphology, osteoclast differentiation, and HIF-1α expression (Western blot). RESULTS:Alamar blue cell viability assay showed no toxicity to RAW cells with the use of velutin in all concentrations tested (p>0.05). Velutin did not induce cell apoptosis based on caspase 3/7 assay (p>0.05). Fluorescence images stained by DAPI showed no alteration in the morphology of RAW cell nuclei (p>0.05) treated with velutin. TRAP assays demonstrated a dose-dependent reduction in osteoclast formation by velutin when compared with control (p<0.05). Velutin showed a reduction in HIF-1α expression related to IκB phosphorylation when compared with control (p<0.001). CONCLUSIONS:At the tested concentrations, velutin was not cytotoxic to RAW 264.7 and differentiated cells. Velutin reduced osteoclast differentiation and downregulated HIF-1α through the NF-κB pathway. 10.1016/j.archoralbio.2017.09.001
    Proliferative and Anti-Inflammatory Effects of Resveratrol and Silymarin on Human Gingival Fibroblasts: A View to the Future. Shahidi Minoo,Vaziri Farzaneh,Haerian Ahmad,Farzanegan Amir,Jafari Soudeh,Sharifi Roya,Shirazi Fatemeh Sadeghi Journal of dentistry (Tehran, Iran) OBJECTIVES:It has been demonstrated that polyphenol components such as silymarin and resveratrol have anti-inflammatory properties. Periodontitis is a chronic inflammatory disease that leads to the breakdown of dental supporting tissues and tooth loss. The purpose of this study was to investigate the anti-inflammatory effects of silymarin and resveratrol on lipopolysaccharide (LPS)-induced inflammatory response in human gingival fibroblasts (HGFs). MATERIALS AND METHODS:HGFs were treated with different concentrations of silymarin and/or resveratrol (25, 50, 100 and 200μg/ml). The effects of silymarin and resveratrol on cell viability and proliferation were assessed by MTT assay and cell cycle analysis, respectively. Also, HGFs were treated with silymarin and/or resveratrol and were stimulated with LPS. The levels of Interleukin-6 (IL-6) and IL-8 were assessed by enzyme-linked immunosorbent assay (ELISA). RESULTS:After treatment with silymarin, the viability of fibroblasts significantly increased, whereas treatment with resveratrol did not have any significant effect on cell viability. However, the combination of these flavonoids (50μg/ml silymarin and 100μg/ml resveratrol) significantly increased the viability of fibroblasts. Resveratrol significantly inhibited LPS-induced IL-6 and IL-8 secretion by HGFs, but silymarin did not show such a significant effect. CONCLUSIONS:The findings of the present study demonstrated the anti-inflammatory effects of resveratrol and its combination with silymarin. Therefore, the combination of silymarin and resveratrol may be useful as a therapeutic agent for treatment of periodontal diseases.
    Resveratrol Suppresses Matrix Metalloproteinase-2 Activation Induced by Lipopolysaccharide in Mouse Osteoblasts via Interactions with AMP-Activated Protein Kinase and Suppressor of Cytokine Signaling 1. Yu Yaqiong,Li Xiaolin,Mi Jing,Qu Liu,Yang Di,Guo Jiajie,Qiu Lihong Molecules (Basel, Switzerland) () lipopolysaccharide (LPS) is associated with the progression of bone resorption in periodontal and periapical diseases. Matrix metalloproteinase-2 (MMP-2) expression and activity are elevated in apical periodontitis and have been suggested to participate in bone resorption. Therefore, inhibiting MMP-2 activation may be considered a therapeutic strategy for treating apical periodontitis. Resveratrol is a natural non-flavonoid polyphenol that has been reported to have antioxidant, anti-cancer, and anti-inflammatory properties. However, the capacity of resveratrol to protect osteoblast cells from LPS insults and the mechanism of its inhibitory effects on MMP-2 activation is poorly understood. Here, we demonstrate that cell viability is unchanged when 10 mg L LPS is used, and MMP-2 expression is drastically induced by LPS in a concentration- and time-dependent manner. Twenty micromolar resveratrol did not reduce MC3T3-E1 cell viability. Resveratrol increased AMP-activated protein kinase (AMPK) phosphorylation, and Compound , a specific AMPK inhibitor, partially abolished the resveratrol-mediated phosphorylation of AMPK. In addition, AMPK inhibition blocked the effects of resveratrol on MMP-2 expression and activity in LPS-induced MC3T3-E1 cells. Treatment with resveratrol also induced suppressor of cytokine signaling 1 (SOCS1) expression in MC3T3-E1 cells. SOCS1 siRNA negated the inhibitory effects of resveratrol on LPS-induced MMP-2 production. Additionally, resveratrol-induced SOCS1 upregulation was reduced by treatment with compound . These results demonstrate that AMPK and SOCS1 activation are important signaling events during resveratrol-mediated inhibition of MMP-2 production in response to LPS in MC3T3-E1 cells, and there is crosstalk between AMPK and SOCS1 signaling. 10.3390/molecules23092327
    Mangiferin promotes the osteogenic differentiation of human periodontal ligament stem cells via TGF‑β/SMAD2 signaling. Molecular medicine reports Mangiferin (MAG) is a polyphenolic compound present in mangoes. This compound suppresses inflammation and decreases bone destruction. This study aimed to determine whether MAG directly promotes proliferation and osteogenic differentiation of human periodontal ligament stem cells (hPDLSCs). Cell proliferation and osteogenic differentiation experiments were performed in hPDLSCs, and MAG was used as a stimulator during osteogenic induction. Alkaline phosphatase (ALP) activity and Alizarin red staining were analyzed, and the expression of osteogenesis‑associated genes was investigated by reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR) and western blot analysis to determine the effect of MAG on the osteogenic differentiation of hPDLSCs. Galunisertib was used to selectively inhibit TGF‑β/SMAD2 signaling. Western blotting was performed to study the underlying mechanism. Cell Counting Kit‑8 assay showed that MAG did not promote the proliferation of hPDLSCs. MAG (200 µM) significantly promoted ALP activity, mRNA levels of alkaline phosphatase biomineralization associated, collagen type 1, and runt‑related transcription factor‑2, protein levels of SMAD5, alkaline phosphatase and bone morphogenetic protein 2 protein expression and mineralized nodule formation in hPDLSCs. Furthermore, MAG significantly promoted the phosphorylation of SMAD2. Galunisertib inhibited the activation of SMAD2 and partially reversed the MAG‑mediated promotion of hPDLSC osteogenic differentiation. These data indicated that MAG promoted osteogenic differentiation of hPDLSCs potentially through TGF‑β/SMAD2 signaling. Therefore, MAG may help improve periodontal bone loss. 10.3892/mmr.2022.12782
    Salvianolic Acid C Attenuates LPS-Induced Inflammation and Apoptosis in Human Periodontal Ligament Stem Cells via Toll-Like Receptors 4 (TLR4)/Nuclear Factor kappa B (NF-κB) Pathway. Duan Yan,An Wei,Wu Hongmei,Wu Yunxia Medical science monitor : international medical journal of experimental and clinical research BACKGROUND Periodontitis is a chronic inflammatory disease that causes gingival detachment and disintegration of alveolar bone. Salvianolic acid C (SAC) is a polyphenol compound with anti-inflammatory and antioxidant activities that is isolated from Danshen, a traditional Chinese medicine made from the roots of Salvia miltiorrhiza Bunge. The aim of this study was to investigate the mechanisms of underlying its protective effects and its inhibition effect on inflammation and apoptosis in human periodontal ligament stem cells (hPDLSCs). MATERIAL AND METHODS LPS-induced hPDLSCs, as a model mimicking an inflammatory process of periodontitis in vivo, were established to investigate the therapeutic effect of SAC in periodontitis. The inflammatory cytokines secretion and oxidative stress status were measured by use of specific commercial test kits. The hPDLSCs viability was analyzed by Cell Counting Kit-8 assay. The cell apoptosis and cell cycle were assayed with flow cytometry. Expressions levels of proteins involved in apoptosis, osteogenic differentiation, and TLR4/NF-kappaB pathway were evaluated by Western blotting. Alkaline phosphatase (ALP) activity was detected by ALP assay kit and ALP staining. The mineralized nodules formation of hPDLSCs was checked by Alizarin Red S staining. RESULTS Our results showed that LPS induced increased levels of inflammatory cytokines and oxidative stress and mediated the phosphorylation and nuclear translocation of NF‑kappaB p65 in hPDLSCs. SAC reversed the abnormal secretion of inflammatory cytokines and inhibited the TLR4/NF‑kappaB activation induced by LPS. SAC also upregulated cell viability, ALP activity, and the ability of osteogenic differentiation. The anti-inflammation and TLR4/NF‑kappaB inhibition effects of SAC were reversed by TLR4 overexpression. CONCLUSIONS Taken together, our results revealed that SAC effectively attenuates LPS-induced inflammation and apoptosis via the TLR4/NF-kappaB pathway and that SAC is effective in treating periodontitis. 10.12659/MSM.918940
    Dimethyl fumarate inhibits osteoclasts via attenuation of reactive oxygen species signalling by augmented antioxidation. Yamaguchi Yuuki,Kanzaki Hiroyuki,Katsumata Yuta,Itohiya Kanako,Fukaya Sari,Miyamoto Yutaka,Narimiya Tsuyoshi,Wada Satoshi,Nakamura Yoshiki Journal of cellular and molecular medicine Bone destructive diseases are common worldwide and are caused by dysregulation of osteoclast formation and activation. During osteoclastogenesis, reactive oxygen species (ROS) play a role in the intracellular signalling triggered by receptor activator of nuclear factor-κB ligand (RANKL) stimulation. Previously, we demonstrated that induction of antioxidant enzymes by Nrf2 activation using Nrf2-gene transfer, an ETGE-peptide or polyphenols, successfully ameliorated RANKL-dependent osteoclastogenesis. Dimethyl fumarate (DMF) has been shown to activate Nrf2 signalling and has been lately used in clinical trials for neurodegenerative diseases. In this study, we hypothesized that Nrf2 activation by DMF would inhibit osteoclastogenesis and bone destruction via attenuation of intracellular ROS signalling through antioxidant mechanisms. RAW 264.7 cells were used as osteoclast progenitor cells. We found that DMF induced Nrf2 translocation to the nucleus, augmented Nrf2 promoter-luciferase reporter activity and increased antioxidant enzyme expression. Using flow cytometry, we found that DMF attenuated RANKL-mediated intracellular ROS generation, which resulted in the inhibition of RANKL-mediated osteoclastogenesis. Local DMF injection into the calvaria of male BALB/c mice resulted in attenuated bone destruction in lipopolysaccharide-treated mice. In conclusion, we demonstrated in a preclinical setting that DMF inhibited RANKL-mediated osteoclastogenesis and bone destruction via induction of Nrf2-mediated transcription of antioxidant genes and consequent decrease in intracellular ROS levels. Our results suggest that DMF may be a promising inhibitor of bone destruction in diseases like periodontitis, rheumatoid arthritis and osteoporosis. 10.1111/jcmm.13367
    Resveratrol suppresses the alveolar bone resorption induced by artificial trauma from occlusion in mice. Matsuda Y,Minagawa T,Okui T,Yamazaki K Oral diseases OBJECTIVE:Besides inflammatory bone loss, trauma from occlusion (TO)-induced alveolar bone loss increases the risk of future tooth loss. We have shown that resveratrol, a polyphenol, possesses anti-inflammatory characteristics and a suppressive effect on osteoclastogenesis. Therefore, we investigated the effects of resveratrol on TO-induced bone loss in mice. MATERIAL AND METHODS:Trauma from occlusion was induced by overlaying composite resin onto the maxillary first molar of C57BL/6 mice. TO-induced mice were administered either resveratrol or vehicle for 15 days from 5 days before TO induction. The mice administered vehicle only served as controls. The effect of resveratrol on bone resorption was assessed histologically. Gene expression in gingival and periodontal ligament tissues was analyzed. In vitro effect of resveratrol on the differentiation of RAW 264.7 cells and bone marrow-derived macrophages into osteoclastic cells was analyzed. RESULTS:Resveratrol administration significantly decreased the bone loss and suppressed the elevated expression of osteoclastogenesis-related gene in periodontal ligament tissue by TO. Resveratrol treatment also suppressed the differentiation of both RAW 264.7 cells and bone marrow-derived macrophages into osteoclastic cells. CONCLUSION:Resveratrol administration suppressed the TO-induced alveolar bone loss by suppressing osteoclast differentiation, suggesting that resveratrol is effective in preventing both inflammation and mechanical stress-induced alveolar bone resorption. 10.1111/odi.12785
    Resveratrol enhances the functionality and improves the regeneration of mesenchymal stem cell aggregates. Wang Yi-Jing,Zhao Pan,Sui Bing-Dong,Liu Nu,Hu Cheng-Hu,Chen Ji,Zheng Chen-Xi,Liu An-Qi,Xuan Kun,Pan Ya-Ping,Jin Yan Experimental & molecular medicine Mesenchymal stem cell (MSC)-based regeneration, specifically cell aggregate or cell sheet engineering, is a promising approach for tissue reconstruction. Considering the advantages of ease of harvest and lack of immune rejection, the application of autologous MSCs (i.e., patients' own MSCs) in regenerative medicine has developed considerable interest. However, the impaired cell viability and regenerative potential following MSCs impacted by disease remain a major challenge. Resveratrol (RSV) exhibits reliable and extensive rejuvenative activities that have received increasing clinical attention. Here, we uncovered that resveratrol enhances the functionality and improves the regeneration of mesenchymal stem cell aggregates. Periodontal ligament MSCs (PDLSCs) from normal control subjects (N-PDLSCs) and periodontitis patients (P-PDLSCs) were investigated. Compared to N-PDLSCs, P-PDLSCs were less capable of forming cell aggregates, and P-PDLSC aggregates showed impaired osteogenesis and regeneration. These functional declines could be mimicked in N-PDLSCs by tumor necrosis factor alpha (TNF-α) treatment. Notably, a TNF-α-induced functional decline in N-PDLSC aggregates was rescued by RSV application. More importantly, in both N-PDLSCs and P-PDLSCs, RSV promoted cell aggregate formation and improved their osteogenic potential. Furthermore, as proven ectopically in vivo, the tissue regenerative capability of P-PDLSC aggregates was also enhanced after RSV treatment during aggregate formation in vitro. Finally, in a rat in situ regeneration model, we successfully applied both N-PDLSC aggregates and P-PDLSC aggregates to repair periodontal defects upon long-term functional improvements by RSV preconditioning. Together, our data unravel a novel methodology for using pharmacology (i.e., RSV)-based cell aggregate engineering to improve the functionality and facilitate the regeneration of MSCs from both healthy and inflammatory microenvironments, shedding light on improving the application of autologous MSC-mediated regenerative medicine. 10.1038/s12276-018-0109-y
    Activation of Functional Somatic Stem Cells Promotes Endogenous Tissue Regeneration. Journal of dental research Periodontal ligament derived stem cells (PDLSCs) are capable of differentiating into multiple cell types and inducing a promising immunomodulation for tissue regeneration and disease treatment. However, it is still challenging to develop a practical approach to activate endogenous stem cells for tissue self-healing and regeneration. In this study, transcriptome analysis reveals that resveratrol promotes PDLSC stemness through activation of stem cell, osteoprogenitor, and chondroprogenitor markers. Self-renewal and multipotent differentiation abilities are also improved in resveratrol-treated PDLSCs. In addition, immunomodulation of PDLSCs is dramatically increased after resveratrol treatment. Mechanistically, we show that resveratrol activates ERK/WNT crosstalk through elevation of olfactory and growth factor signaling pathways to upregulate the expression levels of RUNX2 and FASL for osteogenesis and immunomodulation, respectively. By using a periodontitis animal model, administration of resveratrol partially rescues bone loss through activation of endogenous somatic stem cells and inhibition of inflammatory T-cell infiltration. Taken together, our findings identify a novel pharmacological approach to achieve autotherapies for endogenous tissue regeneration. 10.1177/00220345211070222
    Resveratrol and Celastrol Loaded Collagen Dental Implants Regulate Periodontal Ligament Fibroblast Growth and Osteoclastogenesis of Bone Marrow Macrophages. Wang Ruijie,Bao Bin,Bao Chunling,Wang Shujun,Ur Rahman Saeed,Hou Chunyu,Elango Jeevithan,Wu Wenhui Chemistry & biodiversity Collagen is widely used for dental therapy in several ways such as films, 3D matrix, and composites, besides traditional Chinese medicine (TCM), has been used in tissue regeneration and wound healing application for centuries. Hence, the present study was targeted for the first time to fabricate collagen film with TCM such as resveratrol and celastrol in order to investigate the human periodontal ligament fibroblasts (HPLF) growth and bone marrow macrophages (BMM) derived osteoclastogenesis. Further, the physicochemical, mechanical and biological activities of collagen-TCM films crosslinked by glycerol and EDC-NHS (1-ethyl-3-(3-dimethylaminopropyl)carbodiimide-N-hydroxysulfosuccinimide) were investigated. Collagen film characterization was significantly regulated by the nature of plasticizers like hydrophobic and degree of polarity. Interestingly, the collagen film's denaturation temperature was increased by EDC-NHS than glycerol. FT-IR data confirmed the functional group changes due to chemical interaction of collagen with TCM. Morphological changes of HPLF cells cultured in control and collagen films were observed by SEM. Importantly, the addition of resveratrol upregulated the proliferation of HPLF cells, while osteoclastogenesis of BMM cells treated with mCSF-RANKL was significantly downregulated by celastrol. Accordingly, the collagen-TCM film could be an interesting material for dental regeneration, and especially it is a therapeutic target to restrain the elevated bone resorption during osteoporosis. 10.1002/cbdv.202000295
    Resveratrol rescues TNF‑α‑induced inhibition of osteogenesis in human periodontal ligament stem cells via the ERK1/2 pathway. Yuan Jiakan,Wang Xuxia,Ma Dan,Gao Hui,Zheng Dehua,Zhang Jun Molecular medicine reports Periodontitis is a common inflammatory disorder affecting the tissues surrounding the teeth, which can lead to the destruction of periodontal tissue and tooth loss. Resveratrol, a natural phytoalexin, exerts multiple biological effects. For example, its anti‑inflammatory activity has been widely studied for the treatment of inflammatory bowel disease for a number of years. However, its effect on bone repair and new bone formation in an inflammatory microenvironment is not well understood. Accordingly, the effect of resveratrol on inflammation‑affected human periodontal ligament stem cells (hPDLSCs) requires further investigation. In the present study, the effect of tumor necrosis factor‑α (TNF‑α), resveratrol, or the combination of both on the osteogenic differentiation of hPDLSCs, as well as the underlying mechanisms involved, were investigated. Cell Counting Kit‑8 assay, alkaline phosphatase staining, Alizarin red staining, Oil Red O staining, reverse transcription‑quantitative PCR and western blotting were used in the present study. It was demonstrated that resveratrol enhanced hPDLSC osteogenesis and reversed the inhibitory effects of TNF‑α on this process. Further mechanistic studies indicated that resveratrol exerted anti‑inflammatory activity by activating the ERK1/2 pathway, decreasing the secretion of interleukin (IL)‑6 and IL‑8 induced by TNF‑α, and enhancing hPDLSCs osteogenesis. The present study suggested that resveratrol may be a novel and promising therapeutic choice for periodontitis. 10.3892/mmr.2020.11021
    Comparative evaluation of the effect of curcumin and chlorhexidine on human fibroblast viability and migration: An study. Journal of Indian Society of Periodontology BACKGROUND AND OBJECTIVE:Chemical plaque control acts as an adjunct to mechanical periodontal therapy. Chlorhexidine (CHX) is considered as the gold standard in chemical plaque control, but the main concern is about its fibroblast cytotoxicity. Curcumin, a lipophilic polyphenol, may offer as a promising antiplaque agent. This study was conducted to compare the effect of curcumin (0.003%, 0.03%, 0.06%, 0.1%, and 0.12%) and CHX (0.03%, 0.06%, 0.1%, 0.12%, and 0.2%) on gingival fibroblast cell viability and wound healing at different time periods (1, 2, 4, 6, 8, and 10 min). MATERIALS AND METHODS:The minimum inhibitory concentration (MIC50) was determined before the evaluation of cytotoxicity and wound healing property. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and morphological examination by direct invert microscopy were carried out to determine cytotoxicity. Wound healing was evaluated by scratch wound assay. RESULTS AND DISCUSSION:The MIC50 of CHX and curcumin was at 0.1% and 0.003%, respectively. The mean percentage of fibroblast viability at different concentrations of CHX and curcumin at each time period showed a significant difference. Curcumin exhibited less cytotoxicity as compared to CHX at all concentrations and at varying time periods. There was a significant difference between mean percentage of fibroblast viability at MIC50 of CHX (0.1%) and curcumin (0.003%) at different time periods. The difference between percentage wound healing at antibacterial concentrations of CHX and curcumin at varying time periods was significant. CONCLUSION:The antibacterial concentration of curcumin (0.003%) exhibits less fibroblast cytotoxicity and excellent wound healing property as compared to CHX. Curcumin may offer as a promising chemical plaque control agent which is less cytotoxic, cost-effective, safe, easily available, and with a possibly beneficial effect on wound healing. 10.4103/jisp.jisp_173_19
    Radical-scavenging and Pro-/anti-inflammatory Activity of Tetracycline and Related Phenolic Compounds With or Without Visible Light Irradiation. Murakami Yukio,Kawata Akifumi,Suzuki Seiji,Fujisawa Seiichiro In vivo (Athens, Greece) BACKGROUND/AIM:Microbial tetracycline (TC) pastes have been employed to treat oral bacterial infection. In the present study, we investigated the kinetic radical-scavenging and pro-/anti-inflammatory activity of TC with or without visible light irradiation (VLI). MATERIALS AND METHODS:The radical-scavenging activity of TC and minocycline (MC) was determined by differential scanning calorimetry (DSC). The stoichiometric factor (n) and the rate constant of inhibition and propagation (k/k) were determined. The levels of cyclooxygenase-2 (Cox2), tumor necrosis factor-α (Tnfα) or nitric oxide synthase 2 (Nos2) mRNA in RAW264.7 cells stimulated with lipopolysaccharide (LPS) were investigated using real-time reverse transcriptase-polymerase chain reaction. RESULTS:The n and k/k values for 1 mM TC in 2,2'-azobisisobutyronitrile and benzoyl peroxide systems were 0.1-0.2 and 119-250, respectively, whereas the corresponding values for quercetin (QU) and resveratrol (RE) were 2-4 and 7-15, respectively. In RAW264.7 cells stimulated with LPS, Cox2 and Tnfα mRNA were over-expressed in the presence of TC. MC down-regulated only the expression of Cox2 by about 50% in LPS-stimulated cells. The anti-inflammatory activity determined on the basis of Cox2 inhibition declined in the order QU>RE>MC>TC. Upon application of VLI, only TC down-regulated the expression of LPS-stimulated Cox2 and Tnfα mRNA. After exposure to VLI, TC, but not MC, markedly up-regulated hemoxygenase-1 (Ho-1) expression. CONCLUSION:TC is a chain-breaking antioxidant with a large k Upon activation by VLI, TC may undergo degradation and its degradation products affect pleiotropic mediators such as Cox2, Tnfα and Ho-1. TC may be useful as a local photodynamic therapy for periodontal diseases. 10.21873/invivo.11748
    The Effect of Liposomal Curcumin as an Anti-Inflammatory Strategy on Lipopolysaccharide e from Treated Endothelial Committed Neural Crest Derived Stem Cells: Morphological and Molecular Mechanisms. Diomede Francesca,Fonticoli Luigia,Guarnieri Simone,Della Rocca Ylenia,Rajan Thangavelu Soundara,Fontana Antonella,Trubiani Oriana,Marconi Guya Diletta,Pizzicannella Jacopo International journal of molecular sciences Curcumin, a yellow polyphenol extracted from the turmeric root is used as a diet supplement. It exhibits anti-inflammatory, antioxidant, and antitumor properties by modulating different intracellular mechanisms. Due to their low solubility in water, the curcumin molecules must be encapsulated into liposomes to improve the bioavailability and biomedical potential. For the periodontal tissue and systemic health, it is essential to regulate the local inflammatory response. In this study, the possible beneficial effect of liposomes loaded with curcumin (CurLIP) in neural crest-derived human periodontal ligament stem cells (hPDLSCs) and in endothelial-differentiated hPDLSCs (e-hPDLSCs) induced with an inflammatory stimulus (lipopolysaccharide obtained from , LPS-G) was evaluated. The CurLIP formulation exhibited a significant anti-inflammatory effect by the downregulation of Toll-like receptor-4 (TLR4)/Myeloid differentiation primary response 88 (MyD88)/nuclear factor kappa light chain enhancer of activated B cells (NFkB)/NLR Family Pyrin Domain Containing 3 (NLRP3)/Caspase-1/Interleukin (IL)-1β inflammation cascade and reactive oxygen species (ROS) formation. Moreover, the exposure to LPS-G caused significant alterations in the expression of epigenetic modifiers, such as DNA Methyltransferase 1 (DNMT1) and P300, while the CurLIP treatment showed physiological expression. Overall, our in vitro study provides novel mechanistic insights into the intracellular pathway exert by CurLIP in the regulation of inflammation and epigenetic modifications. 10.3390/ijms22147534
    Grafting resveratrol onto mesoporous silica nanoparticles towards efficient sustainable immunoregulation and insulin resistance alleviation for diabetic periodontitis therapy. Journal of materials chemistry. B Host-modulation therapy is generally accepted as a novel promising method for diabetic periodontitis (DP) treatment and screening an appropriate drug model is the key to success. Resveratrol (RSV), because of its viable antioxidative and anti-inflammatory properties and its ability to control glucose metabolism, is considered a potential candidate. However, poor water solubility, rapid decomposition and short serum half-life period significantly limit its application. Therefore, in this study, we designed a RSV-grafted mesoporous silica nanoparticle (MSN-RSV) drug carrier system to enhance RSV's stability effectively and prolong its duration. Further analyses have verified the indispensable role of MSNs in improving the bioavailability of RSV, which could result in a more favorable therapeutic efficacy in DP related to regulating the polarization of the macrophage. The reason for this could be explained by activating the SIRT1/AMPK signaling pathway and inhibiting the NF-B signaling pathway. This study also focused on the auxiliary effect of MSN-RSV on alleviating insulin resistance (IR) and controlling glucose metabolism. In brief, the study has provided a potential alternative strategy for DP therapy. It is also helpful for future intensive research topics like the immunoregulatory mechanisms in the bidirectional relationship between diabetes and periodontitis. 10.1039/d2tb00484d
    The Impact of Resveratrol Supplementation on Blood Glucose, Insulin, Insulin Resistance, Triglyceride, and Periodontal Markers in Type 2 Diabetic Patients with Chronic Periodontitis. Zare Javid Ahmad,Hormoznejad Razie,Yousefimanesh Hojat Allah,Zakerkish Mehrnoosh,Haghighi-Zadeh Mohammad Hosein,Dehghan Parvin,Ravanbakhsh Maryam Phytotherapy research : PTR The aim of this study was to investigate the impact of resveratrol supplementation along with non-surgical periodontal treatment on blood glucose, insulin, insulin resistance, triglyceride (TG), and periodontal markers in patients with type 2 diabetes with periodontal disease. In this double-blind clinical trial study, 43 patients with diabetes with chronic periodontitis were participated. Subjects were randomly allocated to intervention and control groups. The intervention and control groups received either 480 mg/day of resveratrol or placebo capsules (two pills) for 4 weeks. Fasting blood glucose, insulin, insulin resistance (homeostasis model assessment of insulin resistance), TGs, and pocket depth were measured in all subjects' pre-intervention and post-intervention. The mean serum levels of fasting insulin and insulin resistance (homeostasis model assessment of insulin resistance) were significantly lower in the intervention group compared with control group (10.42 ± 0.28 and 10.92 ± 0.9; 3.66 ± 0.97 and 4.49 ± 1.56, respectively). There was a significant difference in the mean pocket depth between intervention and control groups (2.35 ± 0.6 and 3.38 ± 0.5, respectively) following intervention. No significant differences were observed in the mean levels of fasting blood glucose and TGs between two groups' post-intervention. It is recommended that resveratrol supplementation may be beneficial as adjuvant therapy along with non-surgical periodontal treatment in insulin resistance and improving periodontal status among patients with diabetes with periodontal disease. Copyright © 2016 John Wiley & Sons, Ltd. 10.1002/ptr.5737
    Anti-inflammatory, anti-osteoclastic, and antioxidant activities of genistein protect against alveolar bone loss and periodontal tissue degradation in a mouse model of periodontitis. Bhattarai Govinda,Poudel Sher Bahadur,Kook Sung-Ho,Lee Jeong-Chae Journal of biomedical materials research. Part A Genistein, a dietary polyphenol primarily found in soy products, has beneficial effects on bone. However, the effect of genistein on inflammatory periodontal destruction has not been investigated in detail. We explored whether genistein protects against lipopolysaccharide (LPS)/ligature-induced periodontitis in mice. We also examined the effect of genistein on LPS-stimulated inflammatory and oxidative stress using RAW 264.7 macrophages and human gingival fibroblasts (hGFs). The results from μCT and histological analyses revealed that intraperitoneal injection of genistein (20 mg/kg body weight) daily for three weeks inhibited LPS-mediated alveolar bone loss and periodontal tissue degradation. The administration of genistein also inhibited osteoclast formation and the expression of inflammation-related molecules in the inflamed region of mice with periodontitis. Treatment with 30-70 μM genistein significantly prevented osteoclast differentiation in receptor activator of nuclear factor κB ligand- or LPS-stimulated macrophages by suppressing the expression of osteoclast-specific molecules. The addition of genistein led to a dose-dependent inhibition of the expression of inflammation-related molecules both in LPS-stimulated macrophages and hGFs. In addition, genistein at 50 μM protected hGFs from LPS-mediated stresses such as mitochondrial impairment and cellular ROS accumulation. However, such protection was significantly diminished by combined treatment with 25 nM bafilomycin A1, a chemical autophagy inhibitor. Collectively, our results indicate that genistein protects against inflammatory periodontal damage by regulating autophagy induction and inhibiting osteoclast activation, the production of inflammation mediators, and mitochondrial oxidative damage. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 2510-2521, 2017. 10.1002/jbm.a.36109
    Resveratrol attenuates the pathogenic and inflammatory properties of Porphyromonas gingivalis. Ben Lagha Amel,Andrian Elisoa,Grenier Daniel Molecular oral microbiology Porphyromonas gingivalis has been strongly associated with chronic periodontitis, which affects tooth-supporting tissues. This Gram-negative anaerobic bacterium produces a repertoire of virulence factors that modulate tissue destruction directly or indirectly by the induction of inflammatory processes. The aim of this study was to investigate the effects of resveratrol, a major polyphenol found in grapes and wine, on the growth and virulence properties of P. gingivalis as well as on gingival keratinocyte tight junction integrity and the host inflammatory response. Resveratrol exhibited antibacterial activity that may result from damage to the bacterial cell membrane. Resveratrol also killed a pre-formed P. gingivalis biofilm and reduced bacterial adherence to matrix proteins. In addition, resveratrol had a protective effect on the integrity of the keratinocyte tight junctions by inhibiting its breakdown by P. gingivalis. This may be related to the ability of resveratrol to inhibit the protease activities of P. gingivalis. Lastly, resveratrol reduced P. gingivalis-mediated activation of the NF-κB signaling pathway and attenuated TREM-1 gene expression as well as soluble TREM-1 secretion in monocytes. The effect on NF-κB activation likely results from the ability of resveratrol to act as a PPAR-γ agonist. In summary, the antibacterial, anti-adherence, and antiprotease properties of resveratrol, as well as its ability to protect the gingival keratinocyte barrier and attenuate the inflammatory response in monocytes suggest that it may be a promising novel therapeutic agent for treating periodontal disease. 10.1111/omi.12260
    Catechin ameliorates Porphyromonas gingivalis-induced inflammation via the regulation of TLR2/4 and inflammasome signaling. Lee Hyun Ah,Song Yu Ri,Park Mi Hee,Chung Hae-Young,Na Hee Sam,Chung Jin Journal of periodontology BACKGROUND:Porphyromonas gingivalis is a major periodontopathogen found in patients with chronic periodontitis that can lead to alveolar bone or tooth loss. Interleukin-1β (IL-1β), a proinflammatory cytokine, is most relevant to the pathogenesis of periodontitis. Catechin is one of the main polyphenol compounds found in green tea and possesses a range of health benefits. This study examined the anti-inflammatory effects of catechin in THP-1-derived macrophages infected with P. gingivalis as well as its effects on P. gingivalis-induced periodontitis in a mouse model. METHODS:The cytokine levels and relevant protein expression in THP-1 cells were measured using an enzyme-linked immunosorbent assay and Western blot analysis, respectively. An apoptosis-associated speck-like protein containing a caspase recruitment domain (ASC) pyroptosome formation was measured by confocal laser scanning microscopy. Micro-computed tomography was used to determine the level of bone loss induced by a P. gingivalis oral infection. RESULTS:Catechin attenuated the production of IL-1β by inhibiting pro-IL-1β expression via the downregulation of nuclear factor-κB, p38 mitogen-activated protein kinase, and Toll-like receptor signaling. In addition, catechin inhibited the activation of inflammasomes induced by P. gingivalis, but did not affect the growth of P. gingivalis. Catechin reduced the level of alveolar bone loss in a P. gingivalis-induced periodontitis mouse model. CONCLUSION:Catechin possesses anti-inflammatory properties by reducing the level of IL-1β production, suggesting that it can potentially be used for the prevention and treatment of periodontal inflammation caused by P. gingivalis. 10.1002/JPER.18-0004
    Quebecol Shows Potential to Alleviate Periodontal Tissue Damage and Promote Bone Formation in Models. Cardinal Sébastien,Ben Lagha Amel,Azelmat Jabrane,Grenier Daniel ACS omega Quebecol is a polyphenolic compound initially isolated from Canadian maple syrup in 2011. Recently, our group demonstrated in a macrophage model that quebecol inhibits the secretion of pro-inflammatory cytokines and reduces the activation of the NF-κB transcription factor. In this study, we further explored the therapeutic potential of quebecol against periodontal disease, an inflammatory disorder of bacterial origin affecting tooth-supporting tissues. More specifically, the effects of this natural compound on matrix metalloproteinase (MMP) activity and macrophage secretion, as well as on the mineralization activity of osteoblasts (bone-forming cells), were investigated. Results showed that exposing lipopolysaccharide (LPS)-treated macrophages to quebecol led to a significant decrease in the secretion of MMP-8 and MMP-9. In addition, quebecol dose dependently inhibited the catalytic activity of MMP-9. Quebecol also enhanced the mineralization activity of osteoblasts. This study brought forward additional evidence to support the potential of quebecol as a nutraceutical agent against periodontitis. 10.1021/acsomega.1c03312
    Anti-histaminic Effects of Resveratrol and Silymarin on Human Gingival Fibroblasts. Farzanegan Amir,Shokuhian Mohammad,Jafari Soudeh,Shirazi Fatemeh Sadeghi,Shahidi Minoo Inflammation Periodontitis as a chronic inflammatory disease leads to the destruction of the supportive tissues of affected teeth. Crosstalk between periodontitis and the host immune system plays a crucial role in the pathogenesis of this disease. Since polyphenol components such as silymarin and resveratrol have anti-bacterial and anti-inflammatory effects on periodontal tissues, the purpose of this study was to investigate the anti-histaminic effects of silymarin and resveratrol on human gingival fibroblasts (HGFs). HGFs were treated with a concentration of silymarin or resveratrol (100 μg/ml) and a combination of these two polyphenols (50/100 or 100/200 μg/ml silymarin/resveratrol). The effect of silymarin and resveratrol on cell viability was assessed by MTT assay. Also, HGFs were treated with silymarin and/or resveratrol and were stimulated by histamine. The levels of interleukin-6 (IL-6), interleukin-8 (IL-8), tumor necrosis factor alpha (TNF-α), and tissue plasminogen activator 1 (TPA-1) were assessed by enzyme-linked immunosorbent assay (ELISA). After treatment with silymarin, the viability of fibroblast cells significantly increased, whereas treatment with resveratrol and combinations of these flavonoids (silymarin 50 μg/ml and resveratrol 100 μg/ml) did not have any significant effect on cell viability after 24 h. Treatment with 100/200 μg/ml silymarin/resveratrol significantly decreased the cell viability after 48 h. Resveratrol inhibited histamine-induced IL-6 secretion by HGFs significantly, whereas silymarin showed significant effect on TNF-α. A blend of silymarin and resveratrol displayed more valuable results. In conclusion, combination of resveratrol and silymarin could significantly inhibit inflammatory effects of histamine on cultured HGFs by reduction of IL-6, IL-8, TPA-1, and TNF-α. 10.1007/s10753-019-01023-z
    Resveratrol and insulin association reduced alveolar bone loss and produced an antioxidant effect in diabetic rats. Cirano Fabiano Ribeiro,Molez Andréia Manetta,Ribeiro Fernanda Vieira,Tenenbaum Howard C,Casati Marcio Z,Corrêa Mônica Grazieli,Pimentel Suzana Peres Journal of periodontology BACKGROUND:The present investigation studied the effects of systemic administration of resveratrol (RSV) on the development of experimental periodontitis (EP) and on the release of markers of inflammation, bone metabolism, and oxidative stress in diabetic rats. METHODS:Seventy-five male rats were divided into five groups: DM+PLAC: Diabetes Mellitus + placebo solution; DM+INS: DM + insulin therapy; DM+RSV: DM + RSV; DM+RSV+INS: DM + RSV and insulin; NDM: non-diabetic. Streptozotocin was used to induce DM and EP was induced by the placement of a ligature at the fist mandibular and the second maxillary molars. Euthanasia occurred 30 days after the initiation of the study and mandible specimens were subjected for morphometric analysis of bone level. Gingival tissues from mandibular molars were collected for quantification of inflammatory and oxidative stress markers by multiplex assay system and ELISA assay, respectively. Maxillary gingival tissues were processed for real-time polymerase chain reaction (real-time PCR) assessment of markers of bone metabolism and oxidative stress. RESULTS:Morphometric analysis revealed greater bone loss in DM+PLAC and DM+INS in comparison to the other treatments (P < 0.05). RSV used in conjunction with INS reduced the levels of interleukin (IL)-1β, IL-6, IL-17, interferon-gamma (IFN-γ) and superoxide dismutase 1 (SOD) (P < 0.05). RSV alone reduced nicotinamide adenine dinucleotide phosphatase oxidase (NADPH oxidase) levels, in comparison to DM+INS and DM+RSV+INS (P < 0.05). All treatments upregulated mRNA levels for osteoprotegerin (OPG) in comparison to PLAC (P < 0.05). Sirtuin 1 (SIRT) mRNA levels were lower in PLAC when compared to DM+RSV, DM+RSV+INS and NDM (P < 0.05). CONCLUSION:RSV reduced the progression of EP and the levels of NADPH oxidase. Co-treatment with RSV and insulin reduced the levels of pro-inflammatory factors (either proteins or mRNA) and increased the levels of SOD. The data also demonstrated that treatment with RSV and INS alone or in combination had beneficial effects on bone loss. 10.1002/JPER.19-0718
    Inhibitory effect of oolonghomobisflavan B on osteoclastogenesis by suppressing p38 MAPK activation. Lim Soomin,Kim Tae Hoon,Ihn Hye Jung,Lim Jiwon,Kim Gi-Young,Choi Yung Hyun,Bae Jong-Sup,Jung Jae-Chang,Shin Hong-In,Kim Jung-Eun,Park Eui Kyun Bioorganic & medicinal chemistry letters Suppression of differentiation and/or function of osteoclasts is considered an effective therapeutic strategy for osteolytic bone diseases such as periodontitis and osteoporosis. Evidence regarding the health benefits of oolong tea consumption is accumulating, and tea polyphenols have various pharmacological properties such as anti-cancer and anti-diabetes effects. In this study, we investigated the effect of oolonghomobisflavan B (OFB), a polyphenolic compound in oolong tea, on osteoclast differentiation. OFB suppressed receptor activator of nuclear factor-κB (RANKL)-induced formation of tartate-resistant acid phosphatase-positive multinuclear cells without cytotoxicity. OFB also significantly attenuated p38 phosphorylation, which is essential for RANKL-induced osteoclastogenesis, and inhibited the expressions of nuclear factor of activated T cells, cytoplasmic 1 (NFATc1) and osteoclast-specific target genes, including dendritic cell-specific transmembrane protein and cathepsin K. Our findings suggest that OFB exhibits an anti-osteoclastogenic activity by inhibiting RANKL-mediated p38 activation, which is useful for the prevention and treatment of osteolytic bone diseases. 10.1016/j.bmcl.2020.127429
    Impact of resveratrol supplementation on inflammatory, antioxidant, and periodontal markers in type 2 diabetic patients with chronic periodontitis. Javid Ahmad Zare,Hormoznejad Razie,Yousefimanesh Hojat Allah,Haghighi-Zadeh Mohammad Hosein,Zakerkish Mehrnoosh Diabetes & metabolic syndrome BACKGROUND:Diabetes mellitus and periodontal disease are two common and chronic diseases with bidirectional relationship influence public health and quality of life. The aims of this study was to study the impact of resveratrol supplementation in adjunct with non-surgical periodontal therapy on inflammatory, antioxidant, and periodontal markers in patients with type 2 diabetes with periodontal disease. MATERIALS AND METHODS:In this randomized clinical trial, 43 patients with diabetes and chronic periodontitis were randomly allocated into two intervention and control groups receiving either resveratrol supplements or placebo for 4 weeks. Serum levels of interleukin 6 (IL6), tumor necrosis factor α (TNFα), total antioxidant capacity (TAC) and clinical attachment loss (CAL) as the main index of periodontal marker were measured pre-intervention and post-intervention. RESULTS:In the intervention group, the mean serum level of IL6 was reduced significantly (P = 0.039) post-intervention (2.19 ± 1.09 and 1.58 ± 1.06). No significant differences were seen in the mean levels of IL6, TNFα, TAC and CAL between two groups post-intervention. CONCLUSIONS:It is suggested that daily consumption of resveratrol supplement may not change TNFα, TAC and CAL, but it would be beneficial in reducing serum levels of IL6. Therefore, further studies are suggested to investigate the effects of resveratrol supplementation along with NST on periodontal status. 10.1016/j.dsx.2019.07.042
    A potential role for the silent information regulator 2 homologue 1 (SIRT1) in periapical periodontitis. Kudo H,Takeichi O,Hatori K,Makino K,Himi K,Ogiso B International endodontic journal AIM:To investigate the role played by silent information regulator 2 homologue 1 (SIRT1) during angiogenesis of periapical periodontitis. METHODOLOGY:Periapical granulomas were subjected to dual-colour immunofluorescence imaging and real-time polymerase chain reactions assaying the expression levels of SIRT1, vascular endothelial growth factor (VEGF) and VE-cadherin. The association between Ki-67 and SIRT1 expression was also examined. Human umbilical vein endothelial cells (HUVECs) were treated with a combination of lipopolysaccharide and resveratrol (a SIRT1 activator) or sirtinol (a SIRT1 inhibitor); and the levels of mRNAs encoding SIRT1, VEGF and VE-cadherin were determined. HUVEC tube formation was assayed in the presence of resveratrol or sirtinol. The Mann-Whitney U-test or the Tukey-Kramer test was used for statistical analysis. RESULTS:Ki-67-expressing cells, including endothelial cells, lay adjacent to SIRT1-expressing cells in periapical granulomas. In addition, SIRT1-expressing cells were detected adjacent to VEGF-expressing cells and VEGF- or VE-cadherin-expressing endothelial cells. SIRT1, VEGF and VE-cadherin mRNA expression levels in periapical granulomas were significantly higher (P = 0.0054, 0.0090 and 0.0090, respectively) than those in healthy gingival tissues. HUVECs treated with resveratrol exhibited significantly higher expression of mRNAs encoding SIRT1, VEGF and VE-cadherin (P = 0.0019, 0.00005 and 0.0045, respectively) compared with controls, but sirtinol inhibited such expression. Resveratrol caused HUVECs to form tube-like structures, whilst sirtinol inhibited this process. CONCLUSIONS:These findings suggest that SIRT1 may stimulate angiogenesis in periapical granulomas by triggering the proliferation of endothelial cells and inducing VEGF and VE-cadherin expression. 10.1111/iej.12894
    Pterostilbene complexed with cyclodextrin exerts antimicrobial and anti-inflammatory effects. Lim Yi Rong Ivan,Preshaw Philip M,Lim Lum Peng,Ong Marianne Meng Ann,Lin Hai-Shu,Tan Kai Soo Scientific reports Resveratrol (RES) is a natural polyphenol with potential as an adjunctive therapeutic modality for periodontitis. However, its inferior pharmacokinetics and toxicity concerns about its commonly used solvent dimethyl sulfoxide (DMSO) hinder translation to clinical applicability. Our study aimed to investigate the comparative antimicrobial properties of RES and its analogues (pterostilbene [PTS], oxyresveratrol [OXY] and piceatannol [PIC]), utilizing 2-hydroxypropyl-β-cyclodextrin (HPβCD) as a solubiliser, which has a well-documented safety profile and FDA approval. These properties were investigated against Fusobacterium nucleatum, a key periodontal pathogen. PTS demonstrated the most potent antibacterial effects in HPβCD, with MIC > 60-fold lower than that of RES, OXY and PIC. In addition, PTS inhibited F. nucleatum biofilm formation. PTS exerted antimicrobial effects by eliciting leakage of cellular contents, leading to loss of bacterial cell viability. PTS also conferred immunomodulatory effects on F. nucleatum-challenged macrophages via upregulation of antioxidant pathways and inhibition of NF-κB activation. Given the superior antimicrobial potency of PTS against F. nucleatum compared to RES and other analogues, and coupled with its immunomodulatory properties, PTS complexed with HPβCD holds promise as a candidate nutraceutical for the adjunctive treatment of periodontitis. 10.1038/s41598-020-66031-8
    Effects of theaflavins on tissue inflammation and bone resorption on experimental periodontitis in rats. Wu Ya-Hsin,Kuraji Ryutaro,Taya Yuji,Ito Hiroshi,Numabe Yukihiro Journal of periodontal research BACKGROUND AND OBJECTIVE:Theaflavins (TFs), the major polyphenol in black tea, have the ability to reduce inflammation and bone resorption. The aim of this study was to evaluate the effects of TFs on experimental periodontitis in rats. MATERIAL AND METHODS:Thirty rats were divided into five groups: Control (glycerol application without ligation), Ligature (glycerol application with ligation), TF1 (1 mg/mL TF application with ligation), TF10 (10 mg/mL TF application with ligation), and TF100 (100 mg/mL TF application with ligation). To induce experimental periodontitis, ligatures were placed around maxillary first molars bilaterally. After ligature placement, 100 μL glycerol or TFs were topically applied to the rats daily, and rats were euthanized 7 days after ligature placement. Micro-computed tomography was used to measure bone resorption in the left side of the maxilla, and quantitative polymerase chain reaction was used to measure the expression of interleukin (IL)-6, growth-regulated gene product/cytokine-induced neutrophil chemoattractant (Gro/Cinc-1, rat equivalent of IL-8), matrix metalloproteinase-9 (Mmp-9), receptor activator of nuclear factor-kappa Β ligand (Rankl), osteoprotegerin (Opg), and the Rankl/Opg ratio in gingival tissue. With tissue from the right side of the maxilla, hematoxylin and eosin staining was used for histological analysis, immunohistochemical staining for leukocyte common antigen (CD45) was used to assess inflammation, and tartrate-resistant acid phosphatase (TRAP) staining was used to observe the number of osteoclasts. RESULTS:The TF10 and TF100 groups, but not the TF1 group, had significant inhibition of alveolar bone loss, reduction in inflammatory cell infiltration in the periodontium, and significantly reduced numbers of CD45-positive cells and TRAP-positive osteoclasts compared with the Ligature group. Correspondingly, the TF10 and TF100 groups had significantly downregulated gene expression of IL-6, Gro/Cinc-1(IL-8), Mmp-9, and Rankl, but not of Opg. Consequently, Rankl/Opg expression was significantly increased in the Ligation group but was attenuated in the TF10 and TF100 groups. CONCLUSION:The results of this study suggest that topical application of TFs may reduce inflammation and bone resorption in experimental periodontitis. Therefore, TFs have therapeutic potential in the treatment of periodontal disease. 10.1111/jre.12600
    The histopathological and morphometric investigation of the effects of systemically administered humic acid on alveolar bone loss in ligature-induced periodontitis in rats. Çalışır M,Akpınar A,Poyraz Ö,Göze F,Çınar Z Journal of periodontal research BACKGROUND:Humic acid is a soil extract found widely around the world. This product includes some trace elements important for human's health. The purpose of this study was to evaluate the morphometric and histopathological changes associated with an experimental periodontitis model in rats in response to systemic administration of humic acid. MATERIAL AND METHODS:Thirty-eight male Wistar rats were divided into five experimental groups: non-ligated (NL, n = 6) group; ligature-only (LO, n = 8) group; ligature + systemic administration of humic acid (20, 80 and 150 mg/kg body weight per day for 15 d respectively) (S-20, S-80 and S-150) groups. 4/0 silk ligatures were placed at the gingival margin of lower first molars of the mandibular quadrant. The animals were killed at the end of 15 d. Changes in alveolar bone levels were clinically measured, using a stereomicroscope (× 25), as the distance from the cementoenamel junction to the alveolar bone crest. Tissues were histopathologically examined to assess the differences of osteoclast numbers, osteoblastic activity and inflammatory cell infiltration among the study groups. Enzyme-linked immunosorbent assay interleukin (IL)-1β and IL-10 levels in serum and gingival homogenates were evaluated. RESULTS:At the end of 15 d, the alveolar bone loss was significantly higher in the LO group compared to the NL, S-80 and S-150 groups (p < 0.05). In addition, the alveolar bone loss in the S-80 group was significantly lower than the LO and S-20 groups (p < 0.05). The osteoblastic activity in the S-80 and S-150 groups was significantly higher than the other groups (p < 0.05). The osteoclast number in the LO group was significantly higher than the NL, S-80 and S-150 groups (p < 0.05). Inflammatory cell infiltration was significantly higher in LO and S-20 groups than the other groups (p < 0.05). The highest serum and gingival homogenate IL-10 levels were determined in the S-80 group (p < 0.05). The serum and gingival homogenate IL-1β levels in the LO group were significantly higher than the other groups (p < 0.05). Both 80 and 150 mg/kg dosages of humic acid significantly reduced the periodontitis-related bone loss and inflammation, but the differences between these two groups were not statistically significant (p > 0.05). CONCLUSIONS:Within the limits of this study, it can be suggested that humic acid, when administered systemically as an 80 mg/kg dose, may prevent alveolar bone loss and reduce inflammation in the rat model. 10.1111/jre.12329
    Humic Acid, a Polyphenolic Substance, Decreases Alveolar Bone Loss in Experimental Periodontitis in Rats. Çalışır Metin,Akpınar Aysun,Poyraz Ömer,Göze Fahrettin,Çınar Ziynet Journal of veterinary dentistry The purpose of this study was to evaluate the biochemical, morphometric, and histopathological changes associated with experimental periodontitis in rats in response to local administration of humic acid. Thirty-eight Wistar rats were divided into 5 experimental groups: nonligated (NL) group, ligature-only (LO) group, and ligature + local administration of humic acid (20, 80, and 150 mg/kg body weight per day for 15 days, respectively; L-20, L-80, and L-150 groups). Changes in alveolar bone levels were clinically measured as the distance from the cementoenamel junction to the alveolar bone crest with a stereomicroscope. Tissues were histopathologically examined to assess the osteoclast numbers, osteoblastic activity, and inflammatory cell infiltration among the study groups. Enzyme-linked immunosorbent assay interleukin1β (IL-1β) and IL-10 levels in serum and gingival homogenates were evaluated. At the end of 15 days, the alveolar bone loss was significantly higher in the LO group compared to the NL, L-20, and L-150 groups ( < .05). The osteoclast number in the LO group was significantly higher than the NL, L-20, and L-150 groups ( < .05). Inflammatory cell infiltration was significantly higher in the LO and L-80 groups than the other groups ( < .05). The highest serum and gingival homogenate IL-10 levels were determined in the NL group ( < .05). The serum and gingival homogenate IL-1β levels in LO group were significantly higher than the NL, L-20, and L-150 groups ( < .05). Within the limits of this study, it can be suggested that humic acid, when administered locally at 20 and 80 mg/kg doses, may prevent alveolar bone loss in the rat model. 10.1177/0898756420910531
    Resveratrol decreases local inflammatory markers and systemic endotoxin in patients with aggressive periodontitis. Medicine BACKGROUND:Inflammation is hypothesized to contribute to the pathogenesis of periodontitis. Resveratrol (RV) is known for its anti-inflammatory properties. The purpose of this study was to investigate the inhibitory effect of RV on local inflammatory markers and systemic endotoxin in patients with periodontitis. METHODS:A total of 160 patients with periodontitis were enrolled in this study. The selected patients were randomly divided into four groups and received placebo, high-dose (500 mg/d) of RV (HRV, n = 40), middle-dose (250 mg/d) of RV (middle dose RV (MRV), n = 40) and low-dose (125 mg/d) of RV (low dose RV (LRV), n = 40) with orally administration. All patients received an 8-week treatment. The periodontal status of patients with periodontitis was recorded by using clinical attachment level (CAL), bleeding index (BI), oral hygiene index-simplified (OHI-S), and probing pocket depth (PPD). The levels of inflammatory markers in serum and gingival crevicular fluid (GCF), and systemic levels of endotoxin were evaluated using high sensitivity enzyme-linked immuno sorbent assay. RESULTS:Outcomes showed that symptoms of periodontitis determined by CAL, BI OHI-S and PPD were improved by RV compared to placebo. RV treatment decreased inflammatory markers in serum and GCF compared to placebo in patient with periodontitis. Systemic endotoxin declined more in the RV group than the placebo-treated group. CONCLUSION:In conclusion, data in the current study indicate that RV is an efficient drug for the treatment of patients with periodontitis. The findings of the present study find that RV inhibits systemic local inflammatory markers and systemic endotoxin and suggest that 500 mg/d RV is the ideal dose for patients with periodontitis. 10.1097/MD.0000000000029393
    Mangiferin ameliorates Porphyromonas gingivalis-induced experimental periodontitis by inhibiting phosphorylation of nuclear factor-κB and Janus kinase 1-signal transducer and activator of transcription signaling pathways. Li H,Wang Q,Ding Y,Bao C,Li W Journal of periodontal research BACKGROUND AND OBJECTIVE:Mangiferin is a natural polyphenol compound with anti-inflammatory properties. However, there have been few reports on the effect of mangiferin on periodontitis. Here, we investigated the anti-inflammatory effects of this compound on experimental periodontitis and the underlying mechanisms. MATERIAL AND METHODS:Mice were inoculated with Porphyromonas gingivalis to induce periodontitis, and treated with mangiferin orally (50 mg/kg bodyweight, once a day) for 8 wk. Then, the alveolar bone loss was examined using a scanning electronic microscope. Expression of tumor necrosis factor-α (TNF-α) and the phosphorylation levels of nuclear factor-κB (NF-κB) and Janus kinase 1-signal transducer and activator of adhesion (JAK1-STAT) pathways in the gingival epithelium were detected using western blot analysis and immunohistochemical staining. RESULTS:The results showed that mice with periodontitis exhibited greater alveolar bone loss, stronger expression of TNF-α and higher phosphorylation levels of NF-κB and JAK1-STAT1/3 pathways in gingival epithelia, compared with control mice with no periodontitis. Moreover, treatment with mangiferin could significantly inhibit alveolar bone loss, TNF-α production and phosphorylation of NF-κB and JAK1-STAT1/3 pathways in gingival epithelia. CONCLUSION:Mangiferin has anti-inflammatory effects on periodontitis, which is associated with its ability to down-regulate the phosphorylation of NF-κB and JAK1-STAT1/3 pathways in gingival epithelia. 10.1111/jre.12360
    Systemic treatment with resveratrol reduces the progression of experimental periodontitis and arthritis in rats. Corrêa Mônica G,Pires Paula Rodrigues,Ribeiro Fernanda Vieira,Pimentel Suzana Peres,Cirano Fabiano Ribeiro,Napimoga Marcelo Henrique,Casati Marcio Z,Casarin Renato Corrêa Viana PloS one Rheumatoid arthritis and periodontitis are chronic inflammatory diseases which has been closely associated due to the nature of immune-inflammatory imbalance response. Resveratrol is a naturall product with biological proprieties that may promote immunomodulatory effects on host response. This study investigated resveratrol continuous administration effect on experimental periodontitis and arthritis progression in rats. Thirty-five rats were assigned to the following groups: 1-experimental arthritis + experimental periodontitis + placebo (RA+EP +PL) (n = 12); 2 -RA+EP+ ibuprofen (RA+PE+IB) (n = 11); 3-RA+EP+ resveratrol (RA+PE+RSV) (n = 11). After euthanasia, the specimens were processed for morphometric analysis of bone loss, and the gingival tissue surrounding the first molar was collected for quantification of inflammatory markers using a Luminex/MAGpix assay and anti-citrullinated protein antibody (ACCPA) levels were measured by ELISA assay. Serum level of rheumatoid factor (RF) was measured by ELISA assay. Paw edema was analyzed using a plethysmometer. Higher bone loss was observed in PL group, when compared to IB and RSV groups. RSV group presented higher IL-4 concentration than PL and IB groups. Resveratrol reduced RF serum levels and both IB and RSV decreased ACCPA gingival levels. Besides, paw swelling level was significantly lower in IB and RSV groups in the 21th day and only in RSV group in the 28th day. Histological analyzes showed smooth articular surface and higher width of the subchondral cortical in RSV group. Resveratrol showed modulatory effect and seems to reduce the inflammatory signs of arthritis and articular damage throughout the time. 10.1371/journal.pone.0204414
    Resveratrol attenuates oxidative stress during experimental periodontitis in rats exposed to cigarette smoke inhalation. Corrêa Mônica Grazieli,Absy Samir,Tenenbaum Howard,Ribeiro Fernanda Vieira,Cirano Fabiano Ribeiro,Casati Marcio Z,Pimentel Suzana Peres Journal of periodontal research OBJECTIVES:This study aimed at investigating the effect of the systemic administration of resveratrol (RESV) on oxidative stress during experimental periodontitis in rats subjected to cigarette smoke inhalation. MATERIAL AND METHODS:Experimental periodontitis (EP) was induced in 26 male Wistar rats by the insertion of a ligature around one of the first mandibular and maxillary molars. The animals were assigned randomly to the following groups: cigarette smoke inhalation (CSI; 3 times/d, 8 minutes/d) + resveratrol (10 mg/Kg), that is, SMK + RESV (n = 13) and cigarette smoke inhalation + placebo, that is, SMK + PLAC (n = 13). The substances were administered daily for 30 days (19 days prior and 11 days following EP induction), and then, the animals were euthanized. The maxillary specimens were processed for morphometric analysis of bone loss, and the tissue surrounding the first maxillary molars was collected for mRNA quantification of Sirtuin 1 (SIRT1) by real-time PCR. The gingival tissues surrounding the mandibular first molars were collected for quantification of superoxide dismutase 1 (SOD1) and nicotinamide adenine dinucleotide phosphatase oxidase (NADPH) using an ELISA assay. RESULTS:Reduced bone loss was demonstrated in animals in the SMK + RESV group as compared to those in the SMK + PLAC (P < 0.05) group on the basis of morphometric analysis. Resveratrol promoted higher levels of SIRT and SOD (P < 0.05) as well as reduced levels of NADPH oxidase (P < 0.05) were found in tissues derived from animals in the SMK + RESV group when compared to those in the SMK + PLAC group. CONCLUSION:Resveratrol is an efficient therapeutic agent that reduces exacerbation of bone loss found in animals with EP that were also exposed to smoke. The results suggest that its effects could be mediated, at least in part, by its antioxidant and anti-inflammatory properties which attenuate the effects of oxidative stress on EP in the presence of cigarette smoke. 10.1111/jre.12622
    Green tea polyphenols enhance gingival keratinocyte integrity and protect against invasion by Porphyromonas gingivalis. Lagha Amel Ben,Groeger Sabine,Meyle Joerg,Grenier Daniel Pathogens and disease The gingival epithelium, a stratified squamous tissue that acts as an interface between the external environment and the underlying connective tissue, plays an active role in maintaining periodontal health. The aim of the present study was to investigate the ability of green tea catechins to enhance gingival epithelial barrier function and protect against the disruption of epithelial integrity induced by Porphyromonas gingivalis. Both the green tea extract and epigallocatechin-3-gallate (EGCG) dose- and time-dependently increased the transepithelial electrical resistance (TER) of a gingival keratinocyte model and decreased the permeability of the cell monolayer to fluorescein isothyocyanate-conjugated 4.4-kDa dextran. This was associated with the increased expression of zonula occludens-1 (ZO-1) and occludin, two tight junction proteins. Treating the gingival keratinocyte monolayer with P. gingivalis caused a reduction in TER and affected the distribution of ZO-1 and occludin, allowing P. gingivalis to translocate through the cell monolayer. These deleterious effects mediated by P. gingivalis were abolished by the green tea extract and EGCG. This protection may be in part related to the ability of tea catechins to inhibit the protease activities of P. gingivalis. Given the above properties, green tea catechins may represent promising preventive and therapeutic molecules against periodontal disease. 10.1093/femspd/fty030
    Resveratrol Inhibits Periodontitis-Related Bone Loss in Rats Subjected to Cigarette Smoke Inhalation. Ribeiro Fernanda V,Pino Danilo S,Franck Felipe C,Benatti Bruno B,Tenenbaum Howard,Davies John E,Pimentel Suzana P,Casarin Renato C,Cirano Fabiano R,Casati Marcio Z Journal of periodontology BACKGROUND:Alternative therapeutic approaches have been explored to modulate host response to periodontal disease. Knowledge of new strategies to treat periodontitis is particularly relevant in patients presenting augmented risk to periodontitis, such as smokers. The aim of this study is to investigate the impact of resveratrol (RESV) on progression of experimental periodontitis (EP) in the presence of cigarette smoke inhalation (CSI). METHODS:Rats were assigned to one of three groups: 1) CSI+RESV (n = 20); 2) CSI+placebo (n = 20); and 3) non-CSI (n = 20). CSI was initiated 1 week prior to initiation of RESV or placebo administration (systemically for 30 days) and was continued until the end of the study. EP was induced around the first mandibular and second maxillary molars using ligatures. Specimens from the mandible were processed for morphometric and microcomputed tomography examination of bone volume/levels. Gingival tissues surrounding mandibular molars were collected for quantification of interleukin (IL)-1β, IL-4, IL-6, IL-17, and tumor necrosis factor-α using an assay system. Additional analyses of immunoinflammatory mediator performance (T-helper Type 17 [Th17]/Th2 and Th1/Th2 cell levels) were performed according to Th cell responses in gingival tissues. Gingival tissues of maxillary molars were subjected to real-time polymerase chain reaction for assessment of osteoprotegrin, runt-related transcription factor-2, receptor activator of nuclear factor-kappa B ligand (RANKL), sclerostin, and Dickkopf Wnt signaling pathway inhibitor 1 levels. RESULTS:Higher linear alveolar bone loss (ABL) and lower interradicular bone density were detected in ligated molars in the CSI+placebo group (P <0.05). IL-4 level was the highest, and Th17/Th2 levels were the lowest in RESV-treated rats compared with placebo rats (P <0.05). RESV reduced expression of messenger RNA for RANKL in animals receiving CSI (P <0.05). CONCLUSION:RESV inhibits EP and CSI-induced supporting ABL and has a beneficial effect on osteo-immunoinflammatory markers. 10.1902/jop.2017.170025
    Resveratrol enhances bone formation by modulating inflammation in the mouse periodontitis model. Adhikari Nirpesh,Prasad Aryal Yam,Jung Jae-Kwang,Ha Jung-Hong,Choi So-Young,Kim Ji-Youn,Lee Tae-Hoon,Kim Sang-Hyun,Yamamoto Hitoshi,Suh Jo-Young,An Chang-Hyeon,Lee Youngkyun,Sohn Wern-Joo,An Seo-Young,Kim Jae-Young Journal of periodontal research OBJECTIVE:To evaluate the effect of resveratrol on periodontal bone regeneration after local delivery and to determine its effect on inflammatory mediators. BACKGROUND:Resveratrol is considered an anti-inflammatory polyphenolic stilbene involved in the modulation of inflammation. MATERIALS AND METHODS:Periodontitis was induced in mouse molars using a 5-day ligature model followed by the left second molar extraction and 50 µM resveratrol treatment for 1 and 2 weeks. We then examined specimens treated for 1 week histologically and with immunostaining. Microfocus-computed tomography (micro-CT) was used to examine the bone volume formation. RESULTS:After 1 week of treatment, proinflammatory cytokine levels (TNF-alpha and IL6), cells exhibiting neutrophil and macrophage marker (MPO), cell proliferation marker (Ki67), and preosteoblastic marker (RUNX2) reactivity decreased in the resveratrol-treated specimens compared to the control group. In contrast, we observed a higher number of CD31-, F4/80-, and osteocalcin- (OCN-) positive cells in the resveratrol-treated specimens. After 2 weeks, micro-CT confirmed an increased bone mass in the region of the extraction socket in the resveratrol-treated group. CONCLUSION:After 1 week, the resveratrol-treated specimens revealed evidence of inflammation modulation compared to the control group. These data suggest that resveratrol not only affects inflammation control but also is useful for treating periodontitis-related tissue defects and bone regeneration. 10.1111/jre.12870
    Remodeling immune microenvironment in periodontitis using resveratrol liposomes as an antibiotic-free therapeutic strategy. Shi Junyu,Zhang Yi,Zhang Xiaomeng,Chen Ruiying,Wei Jianxu,Hou Jiazhen,Wang Bing,Lai Hongchang,Huang Yongzhuo Journal of nanobiotechnology BACKGROUND:Periodontitis is a complicated inflammatory disease that damages the tooth-supporting tissues, with limited pharmacotherapy available. Macrophage-targeting therapy is promising for inflammatory diseases. Resveratrol (RSV), a nonflavonoid polyphenol, is known for its anti-inflammatory and immunomodulatory effects. However, its medical application is limited by its poor stability and water-solubility, as well as its low bioavailability. RESULT:A therapeutic resveratrol-loaded liposomal system (Lipo-RSV) was developed to treat periodontitis. The physical properties of Lipo-RSV and its ability to regulate macrophages were investigated. The results showed that Lipo-RSV had good biocompatibility and could re-educate the inflammatory macrophages from M1- to M2-like phenotype through activating p-STAT3 and downregulating p-STAT1. Besides, the Lipo-RSV could scavenge ROS and inhibit the NF-κB signal and inflammasomes, thereby reducing the pro-inflammatory cytokines IL-1β, IL-6, and TNF-α. CONCLUSION:These results revealed that Lipo-RSV could be a potential therapeutic system for the antibiotic-free treatment for periodontal diseases. 10.1186/s12951-021-01175-x
    Effect of red wine or its polyphenols on induced apical periodontitis in rats. Dal-Fabbro Renan,Cosme-Silva Leopoldo,Rezende Silva Martins de Oliveira Fernanda,Capalbo Letícia Cabrera,Plazza Flávia Alfredo,Ervolino Edilson,Cintra Luciano Tavares Angelo,Gomes-Filho João Eduardo International endodontic journal AIM:To evaluate the effect of red wine consumption or its polyphenols on the inflammation/resorption processes associated with apical periodontitis in rats. METHODOLOGY:Thirty-two three-month-old Wistar rats had apical periodontitis induced in four first molars and were then arranged into four groups: control (C)-rats with apical periodontitis; wine (W)-rats with apical periodontitis receiving 4.28 ml/kg of red wine; resveratrol+quercetin (R+Q)-rats with apical periodontitis receiving 4.28 ml/kg of a solution containing 1.00 mg/L of quercetin and 0.86 mg/L of resveratrol and alcohol (ALC)-rats with apical periodontitis receiving the alcoholic dose contained in the wine. The oral gavage treatments were administered daily, from day 0 to day 45. On the 15th day, apical periodontitis was induced, and on the 45th day, the animals were euthanized. Histological, immunohistochemical (RANKL, OPG, TRAP, IL-10, TNF-⍺ and IL-1β) and micro-computed tomography for bone resorption analysis were performed in the jaws. The Kruskal-Wallis with Dunn's test was performed for nonparametric data, and the anova with Tukey's test for parametric data, p < .05. RESULTS:The median score of the inflammatory process was significantly lower in the R+Q group (1) compared to the C (2) (p = .0305) and ALC (3) (p = .0003) groups, and not different from the W (1.5) group. The immunolabeling for OPG was significantly higher in the R+Q group (p = .0054) compared to all groups; the same was observed for IL-10 (p = .0185), different from groups C and ALC. The R+Q group had the lowest TRAP cell count (p < .0001), followed by the W group, both inferior to C and ALC groups. The lowest bone resorption value was in the R+Q group (0.50mm  ± 0.21mm ), significantly lower (p = .0292) than the C group (0.88mm  ± 0.10mm ). The W group (0.60 mm  ± 0.25 mm ) and R+Q group had less bone resorption compared to the ALC group (0.97 mm  ± 0.22 mm ), p = .0297 and p = .0042, respectively. CONCLUSION:Red wine administration to rats for 15 days before induction of apical periodontitis decreased inflammation, TRAP marking and periapical bone resorption compared to alcohol. Resveratrol-quercetin administration reduced the inflammatory process in apical periodontitis, periapical bone resorption, and altered the OPG, IL-10 and TRAP expression compared to C and ALC groups. 10.1111/iej.13633
    2,3,5,4'-Tetrahydroxystilbene-2-O-β-glucoside Isolated from Polygoni Multiflori Ameliorates the Development of Periodontitis. Chin Yu-Tang,Hsieh Meng-Ti,Lin Chi-Yu,Kuo Po-Jan,Yang Yu-Chen S H,Shih Ya-Jung,Lai Hsuan-Yu,Cheng Guei-Yun,Tang Heng-Yuan,Lee Chen-Chen,Lee Sheng-Yang,Wang Ching-Chiung,Lin Hung-Yun,Fu Earl,Whang-Peng Jacqueline,Liu Leroy F Mediators of inflammation Periodontitis, a chronic infection by periodontopathic bacteria, induces uncontrolled inflammation, which leads to periodontal tissue destruction. 2,3,5,4'-Tetrahydroxystilbene-2-O-beta-glucoside (THSG), a polyphenol extracted from Polygoni Multiflori, reportedly has anti-inflammatory properties. In this study, we investigated the mechanisms of THSG on the Porphyromonas gingivalis-induced inflammatory responses in human gingival fibroblasts and animal modeling of ligature-induced periodontitis. Human gingival fibroblast cells were treated with lipopolysaccharide (LPS) extracted from P. gingivalis in the presence of resveratrol or THSG to analyze the expression of TNF-α, IL-1β, and IL-6 genes. Increased AMP-activated protein kinase (AMPK) activation and SirT1 expression were induced by THSG. Treatment of THSG decreased the expression of LPS-induced inflammatory cytokines, enhanced AMPK activation, and increased the expression of SirT1. In addition, it suppressed the activation of NF-κB when cells were stimulated with P. gingivalis LPS. The anti-inflammatory effect of THSG and P. Multiflori crude extracts was reproduced in ligature-induced periodontitis animal modeling. In conclusion, THSG inhibited the inflammatory responses of P. gingivalis-stimulated human gingival fibroblasts and ameliorated ligature-induced periodontitis in animal model. 10.1155/2016/6953459
    Systemic treatment with resveratrol and/or curcumin reduces the progression of experimental periodontitis in rats. Corrêa M G,Pires P R,Ribeiro F V,Pimentel S Z,Casarin R C V,Cirano F R,Tenenbaum H T,Casati M Z Journal of periodontal research BACKGROUND AND OBJECTIVE:Periodontitis is a chronic inflammatory disease of periodontal tissues that leads to the destruction of bone and other connective tissues. Resveratrol and curcumin are plant-derived substances with biological properties that may have immunomodulatory properties. This study investigated the effect of continuous administration of resveratrol and curcumin and the association of resveratrol and curcumin on the progression of experimental periodontitis in rats. MATERIAL AND METHODS:Forty Wistar rats were assigned randomly to the following groups: group 1, experimental periodontitis + placebo (PL) (n = 10); group 2, experimental periodontitis + resveratrol (RSV) (n = 10); group 3, experimental periodontitis + curcumin (C) (n = 10); and group 4, experimental periodontitis + resveratrol + curcumin (COMBI) (n = 10). Periodontitis was induced in rats by tying a silk suture, as a ligature, around one of the first molars. Daily administration of the placebo solution, 10 mg/kg of resveratrol, 100 mg/kg of curcumin or 10 mg/kg of resveratrol plus 100 mg/kg of curcumin was carried out from day 0 to day 30. At the end of the relevant experimental periods, rats were killed and the specimens obtained were processed for morphometric analysis of bone loss. Gingival tissues surrounding the first molar were collected for quantification of interleukin (IL)-1β, IL-4, interferon-gamma (IFN-γ) and tumor necrosis factor-alpha (TNF-α) using a Luminex/MAGPIX assay. RESULTS:Intergroup comparisons of the morphometric outcomes revealed higher bone-loss values in the PL group (p < 0.05) when compared with RSV, C and COMBI groups. There was no difference in bone-loss values among RSV, C and COMBI groups (p > 0.05). The immunoenzymatic assay of the gingival tissue showed a lower concentration of IL-1β in the COMBI group in comparison with the PL group (p < 0.05). Higher values of IL-4 were demonstrated in groups RSV, C and COMBI in comparison with the PL group (p < 0.05). Only RSV caused a reduction in the levels of IFN-γ (p < 0.05). There was no difference in the concentration of TNF-α amongst the four groups (p > 0.05). CONCLUSION:Resveratrol and curcumin are capable of reducing alveolar bone loss in an animal model of periodontitis. This occurred when these agents were added singly or in combination with one another, but there did not appear to be either synergistic or additive effects. 10.1111/jre.12382
    Tea polyphenols protect gingival keratinocytes against TNF-α-induced tight junction barrier dysfunction and attenuate the inflammatory response of monocytes/macrophages. Lagha Amel Ben,Grenier Daniel Cytokine Tea, an aromatic beverage prepared with the leaves of the Camellia sinensis plant, is well known to contain bioactive polyphenols. Green tea contains mainly catechins such as epigallocatechin-3-gallate (EGCG), while black tea is characterized by the presence of theaflavins. TNF-α, which is a pro-inflammatory cytokine that activates the endogenous inflammatory cascade, plays a key role in periodontitis. In the present study, we investigated the ability of tea compounds to attenuate TNF-α-mediated activation of the host inflammatory response in monocytes/macrophages as well as the protective effect of green and black tea polyphenols on gingival keratinocyte barrier dysfunction induced by TNF-α. Tea compounds inhibited both the activation of NF-κB and caspase-1 as well as IL-1β secretion by monocytes/macrophages. TNF-α time-dependently damaged keratinocyte tight junction barrier integrity, as determined by changes in transepithelial electrical resistance and FITC-dextran transport. Green tea extract, EGCG, theaflavins, and to a lesser extent, black tea extract protected keratinocytes against the TNF-α-mediated breakdown of barrier integrity. The treatment of keratinocytes with tea polyphenols markedly mitigated the morphological changes of tight junction proteins such as zonula occludens-1 and occludin compared to cells exposed only to TNF-α, as determined by immunofluorescence. Tea polyphenols also time-dependently decreased the paracellular flux of TNF-α-treated keratinocytes. In conclusion, the ability of tea polyphenols to exert an anti-inflammatory effect and to attenuate the gingival epithelial barrier dysfunction induced by TNF-α supports their potential for the prevention and treatment of periodontal disease. 10.1016/j.cyto.2018.12.009