Calcitonin gene-related peptide enhances osteogenic differentiation and recruitment of bone marrow mesenchymal stem cells in rats.
Jia Sen,Zhang Shi-Jian,Wang Xu-Dong,Yang Zi-Hui,Sun Ya-Nan,Gupta Anand,Hou Rui,Lei De-Lin,Hu Kai-Jin,Ye Wei-Min,Wang Lei
Experimental and therapeutic medicine
The present study evaluated the effects of calcitonin gene-related peptide (CGRP) on bone marrow mesenchymal stem cells (BMMSCs) and in a rat model of mandibular distraction osteogenesis (MDO). Rat BMMSCs were isolated then treated with CGRP or CGRP antagonist (CGRP8-37). The proliferation and migration ability of BMMSCs was determined using 5-bromo-2'-deoxyuridine and Transwell assays, respectively. Osteogenic-related gene expression was analyzed with reverse transcription-quantitative polymerase chain reaction. For the analysis, thirty MDO rats were randomly assigned to control, CGRP or CGRP8-37 groups. To evaluate the mobilization of BMMSCs, nestin and stromal cell-derived factor 1 (SDF-1) were detected by immunohistochemistry and ELISA. Rats were sacrificed following 14 days and new bone formation was assessed by histological and micro-computed tomography analysis. In the results, the CGRP group demonstrated significantly higher migration and proliferation, as well as enhanced alkaline phosphatase and runt-related transcription factor 2 expression compared with the control. In the experiments, bone mineral density of the newly formed bone in the CGRP group was significantly higher than controls. The nestin and SDF-1 expression in the CGRP group was also significantly upregulated. In conclusion, the present study demonstrated that CGRP administration increased new bone formation, possibly via enhancing BMMSC migration and differentiation in MDO rats.
10.3892/etm.2019.7659
Norepinephrine inhibits mesenchymal stem cell chemotaxis migration by increasing stromal cell-derived factor-1 secretion by vascular endothelial cells via NE/abrd3/JNK pathway.
Wu Baolei,Wang Lei,Yang Xi,Mao Ming,Ye Chen,Liu Peng,Yang Zihui,Yang Xinjie,Lei Delin,Zhang Chenping
Experimental cell research
Mesenchymal stem cells (MSCs), which are physiologically maintained in vascular endothelial cell (VEC)-based niches, play a critical role in tissue regeneration. Our previous studies demonstrated that sympathetic denervation could promote MSC mobilization, thereby enhancing bone formation in distraction osteogenesis (DO), a self-tissue engineering for craniofacial and orthopeadic surgeries. However, the mechanisms on how sympathetic neurotransmitter norepinephrine (NE) regulates MSC migration are not well understood. Here we showed that deprivation of NE by transection of cervical sympathetic trunk (TCST) inhibited stromal cell-derived factor-1 (SDF-1) expression in the perivascular regions in rat mandibular DO. In vitro studies showed that NE treatment markedly upregulated p-JNK and therefore stimulated higher SDF-1 expression in VECs than control groups, and siRNA knockdown of the abrd3 gene abolished the NE-induced p-JNK activation. On the other hand, osteoblasts differentiated from MSCs showed an increase in SDF-1 secretion with lack of NE. Importantly, NE-treated VECs inhibited the MSC chemotaxis migration along the SDF-1 concentration gradient as demonstrated in a novel 3-chamber Transwell assay. Collectively, our study suggested that NE may increase the SDF-1 secretion by VECs via NE/abrd3/JNK pathway, thereby inhibiting the MSC chemotaxis migration from perivascular regions toward bone trabecular frontlines along the SDF-1 concentration gradient in bone regeneration.
10.1016/j.yexcr.2016.09.007
Isoquercitrin, a flavonoid glucoside, exerts a positive effect on osteogenesis in vitro and in vivo.
Li Jing,Wang Xuxia,Wang Yingzi,Lu Chengyan,Zheng Dehua,Zhang Jun
Chemico-biological interactions
Isoquercitrin (quercetin-3-O-β-d-glucopyranoside) possess various pharmacological effect as a biologically active compound. The aim of the present study was to investigate its potential effects on the proliferation and osteoblastic differentiation of bone marrow-derived mesenchymal stem cells (BMSCs) in vitro and bone formation in the mid-palatal suture during rapid maxillary expansion (RME) in vivo. Cell proliferation of rat BMSCs was detected by cell-counting kit- 8 (CCK-8) assay. Alkaline phosphatase (ALP) activity assay and alizarin red staining were used to evaluate osteogenic differentiation of BMSCs. Furthermore, the expression levels of osteogenesis-related genes including runt-related transcription factor 2 (Runx2), bone sialoprotein (BSP) and activating transcription factor 6 (ATF6) were measured by real-time RT-PCR. In vivo, thirty-six male 6-week-old Wistar rats with or without suture expansion receive systemic administration of isoquercitrin or saline solution. Micro-CT, HE and Masson staining were used to compare the morphological changes between the groups. Bone morphogenetic protein 2 (BMP2) expression in the suture was detected using immunohistochemical staining. Our results showed that isoquercitrin significantly promoted cell proliferation, ALP activity and mineral deposition in the range from 0.01 to 1 μM. Moreover, the expression levels of Runx2, BSP and ATF6 were also upregulated. The measurement of micro-CT imaging and histological examinations demonstrated that daily oral administration of isoquercitrin (10 mg/kg) increased bone formation compared to the other groups. Furthermore, the expression level of BMP2 was also augmented in the presence of isoquercitrin. Consequently, those findings showed that isoquercitrin exerts stimulatory effects on osteogenesis in vitro and in vivo, suggesting that isoquercitrin could be a potential candidate for preventing relapse following RME within palatal sutures.
10.1016/j.cbi.2018.10.018
Matrine enhances osteogenic differentiation of bone marrow-derived mesenchymal stem cells and promotes bone regeneration in rapid maxillary expansion.
Li Jing,Wang Xuxia,Yang Fan,Yuan Jiakan,Cui Qun,Nie Fujiao,Zhang Jun
Archives of oral biology
OBJECTIVE:The aim of this study was to evaluate the influence of matrine on osteogenic differentiation of rat bone marrow-derived mesenchymal stem cells (BMSCs) as well as on bone metabolism in a rat rapid maxillary expansion (RME) model. METHODS:In in vitro experiments, rat BMSCs were adopted and cell proliferation of BMSCs was measured. Meanwhile, the osteogenic differentiation of BMSCs was detected by alkaline phosphatase (ALP) activity assay, Alizarin red S staining and gene expression. In vivo bone regeneration was analyzed in a rat RME model. Eighteen rats were divided into 3 groups: one group without any treatment, one group receiving only RME, and a group with RME and matrine treatment. After 2 weeks, new bone formation was detected by Micro-CT and histology. Immunohistochemical staining was used to evaluate ALP and BMP2 expression. RESULTS:Overall, we found that matrine upregulated cell proliferation dose-dependently. Also, ALP activity and mineralized matrix generation were enhanced. Moreover, the osteoblast-related gene expression (ALP, bone sialoprotein and osteocalcin) by BMSCs was also promoted. Micro-CT revealed that matrine significantly promoted in vivo bone formation after 2 weeks. Concomitantly, histological examination of haematoxylin-eosin, safranin-O and toluidine blue staining confirmed these findings. In addition, the levels of ALP and BMP2 in the palatal suture tissues of rats with matrine treatment were the highest among three groups. CONCLUSION:This work suggests that matrine regulates osteogenesis and enhances bone regeneration. Matrine treatment may be beneficial in improving the stability of maxillary expansion.
10.1016/j.archoralbio.2020.104862
A preliminary investigation of the effect of relaxin on bone remodelling in suture expansion.
Duarte Carolina,Kobayashi Yukiho,Morita Jumpei,Kawamoto Tatsuo,Moriyama Keiji
European journal of orthodontics
Background and objectives:Relaxin (RLN) is an insulin-like hormone associated with extracellular matrix degradation, osteoclastogenesis, and osteoblast differentiation. This study aimed to assess the effect of RLN during and after lateral expansion of murine calvarial sagittal sutures. Materials and methods:RLN was injected topically using a nano-sized liposome carrier into the sagittal sutures of 8- to 10-week-old wild type mice just before lateral expansion. Suture morphology, bone mineral density (BMD), and bone volume were analysed by micro-computed tomography. Collagen deposition and osteoclast differentiation were observed by Verhoeff-Van Gieson (VVG) and tartrate-resistant acid phosphatase (TRAP) staining, respectively. Results:Less collagen staining and higher tissue-specific relaxin/insulin-like family peptide receptor (Rxfp)-1 and -2 expression were observed in the RLN-treated samples after 48 hours. Increased BMD and volume, and thick well-organised osteoid tissue, with multinucleated TRAP-positive cells, were observed in RLN-treated samples after 1 week. Increased Rxfp-1 expression was observed in the sagittal sutures in the mid-suture fibrous tissue following RLN treatment. Rxfp-2 was only expressed in the calvarial bone under tensile stimulation and RLN treatment further increased its expression. Limitations:RLN-liposomes were not detected at any instance under the current experimental conditions. This is a preliminary study and the sample number limits the power of its results. VVG staining cannot quantify collagen contents but can provide preliminary information on the presence of collagen fibres. Conclusions:RLN treatment may modify bone remodelling and collagen metabolism during and after suture expansion.
10.1093/ejo/cjw037
Effects of curcumin and melatonin on bone formation in orthopedically expanded suture in rats: A biochemical, histological and immunohistochemical study.
Orthodontics & craniofacial research
OBJECTS:To investigate the effects of curcumin (CUR) and melatonin (MEL) on new bone formation following rapid maxillary expansion (RME) in rats. SETTING AND SAMPLE POPULATION:For this study, 24 12-week-old adult male Wistar albino rats from the Animal Laboratory at Adnan Menderes University, Faculty of Medicine, were used. MATERIALS AND METHODS:The rats were randomly divided into the following 3 groups (n = 8 each): only expansion (OE), expansion plus MEL (MEL) and expansion plus CUR (CUR). CUR and MEL were given to the rats during the study period. After the sacrifice of the animals, biochemical, histological and immunohistochemical examinations were performed. RESULTS:Serum bone alkaline phosphatase levels in the MEL group were statistically (P = .007) higher than in the OE group. Serum glutathione peroxidase and catalase activities in the CUR and MEL groups were significantly higher than in the OE group (P = .007 and P = .021, respectively). Inflammatory cell infiltration, new bone formation and capillary intensity parameters did not demonstrate statistically significant differences between the groups (P = .865, P = .067 and P = .055, respectively). The immunohistochemical findings revealed that IL-1, IL-6 and TNF-α H scores showed considerable differences between the groups (all P < .001). The highest IL-1, IL-6 and TNF-α H scores were found in the OE groups rather than in the other groups (P < .001). CONCLUSION:CUR and MEL treatments may be effective in accelerating new bone formation and beneficial in preventing relapse following the RME procedures.
10.1111/ocr.12232
Systemic propolis stimulates new bone formation at the expanded suture: a histomorphometric study.
Altan Burcu A,Kara Isa M,Nalcaci Ruhi,Ozan Fatih,Erdogan Serif M,Ozkut Mahmut M,Inan Sevinc
The Angle orthodontist
OBJECTIVE:To investigate the effects of systemically given propolis on the expanded premaxillary suture in a rat study model. MATERIALS AND METHODS:The 24 rats were randomly divided into three groups-only expansion (OE), expansion plus propolis (PRO), and nonexpansion (control) groups. After the 5-day expansion period was completed, the OE and PRO groups underwent 12 days of mechanical retention. At the end of this period, the animals were euthanatized and their pre-maxillae were dissected and fixed. Histomorphometric examination was performed to determine the number of osteoclasts, osteoblasts, and capillaries as well as the intensity of inflammatory cells and amount of new bone formation. RESULTS:Statistical analysis showed that the intensities of inflammatory cells, number of osteoblasts, and amount of new bone formation were greater in the PRO group than in the other groups. The PRO group also had more osteoclasts and new capillaries. CONCLUSION:Systemic use of propolis may hasten new bone formation at the expanded suture in rats.
10.2319/032612-253.1
Effect of St John's wort on bone formation in the orthopaedically expanded premaxillary suture in rats: a histological study.
Halicioglu Koray,Çörekçi Bayram,Akkaş İsmail,Irgin Celal,Özan Fatih,Yilmaz Fahri,Türker Arzu
European journal of orthodontics
BACKGROUND/OBJECTIVE:The aim of this study was to investigate the effect of systemic St John's wort (Hypericum perforatum) on bone formation in the expanded premaxillary suture in rats. MATERIALS/METHODS:A total of 28 rats were randomly divided into four groups of equal numbers: control (C); only expansion (OE); St John's wort extract given only during the expansion and retention period (a total of 17 days; SJW group); and St John's wort extract given during the nursery phase before expansion (a period of 40 days), and during the expansion and retention periods (a total of 57 days; N + SJW group). After the 5 day expansion period was completed, the rats in the OE, SJW, and N + SJW groups underwent 12 days of mechanical retention, following which they were killed, and their premaxilla dissected and fixed. Histological examination was performed to determine the number of osteoclasts and capillaries, as well as the number of osteoblasts, inflammatory cell infiltration, and the amount of new bone formation. RESULTS:Statistical analysis showed that the number of osteoclasts and capillaries, and the inflammatory cell infiltration, as well as new bone formation, were higher in the SJW and N + SJW groups than in the other groups. However, statistical analysis demonstrated that among these two groups, all parameters, with the exception of the number of capillaries, were higher in the N + SJW group than the SJW group. CONCLUSIONS/IMPLICATIONS:Although more effective in long-term usage, systemic use of St John's wort hastens new bone regeneration at the premaxillary suture and may help prevent relapse after expansion.
10.1093/ejo/cju028