The cytoptrotection of small intestinal submucosa-derived gel in HL-1 cells during hypoxia/reoxygenation-induced injury.
Wang Su-Ya,Sang Jiang-Wei,Ding Wei,Qin Ting-Wu,Bai Lin,Zhang Jie,Luo Jing-Cong
Journal of tissue engineering and regenerative medicine
Small intestinal submucosa (SIS)-derived gel injected into infarcted myocardium has been shown to promote repair and regeneration after myocardial infarction (MI); however, the specific impact of SIS gel on cardiomyocytes remained unknown. The aim of this study was to characterise SIS gel function in hypoxia-reoxygenation (H/R)-induced cardiomyocyte damage and its potential mechanism. HL-1 cardiomyocytes seeded on SIS matrix-coated plates, SIS gel, and uncoated plates were subjected to H/R, cell viability, apoptosis, expression of caspase-3, Bcl-2, and Bax were investigated. SIS gel and SIS matrix as coating substrates markedly improved cell viability, preventing cell apoptosis compared with uncoated plates, with SIS gel yielding the best cytoprotective effects. SIS gel down-regulated expression of pro-inflammatory cytokines (TNF-α, CCL2, and IL-6) by inhibiting the JNK-mitogen-activated protein kinase (MAPK)/NF-κB pathways. Furthermore, SIS gel protected cardiomyocytes from apoptosis by activating protein kinase B (AKT) and extracellular-signal-regulated kinase (ERK) pathways, and markedly up-regulated antiapoptotic Bcl-2 expression but inhibited that of proapoptotic Bax and c-caspase 3. Together, these findings show that SIS gel could decrease H/R-induced cell apoptosis through a mechanism potentially related to its ability to regulate expression of inflammatory cytokines and antiapoptosis signalling pathways to prevent cell apoptosis. Our findings thereby shed light on the mechanism related to SIS gel therapeutic efficacy for MI.
10.1002/term.2878
Human umbilical cord blood-derived mesenchymal stromal cells and small intestinal submucosa hydrogel composite promotes combined radiation-wound healing of mice.
Lee Changsun,Shim Sehwan,Jang Hyosun,Myung Hyunwook,Lee Janet,Bae Chang-Hwan,Myung Jae Kyung,Kim Min-Jung,Lee Seung Bum,Jang Won-Suk,Lee Sun-Joo,Kim Hwi-Yool,Lee Seung-Sook,Park Sunhoo
Cytotherapy
BACKGROUND AIMS:Mesenchymal stromal cells (MSCs) are a promising agent for treating impaired wound healing, and their therapeutic potential may be enhanced by employing extracellular matrix scaffolds as cell culture scaffolds or transplant cell carriers. Here, we evaluated the effect of human umbilical cord blood-derived (hUCB)-MSCs and a porcine small intestinal submucosa (SIS)-derived extracellular matrix scaffold in a combined radiation-wound mouse model of impaired wound healing. METHODS:hUCB-MSCs and SIS hydrogel composite was applied to the excisional wound of whole-body irradiated mice. Assessment of wound closing and histological evaluation were performed in vivo. We also cultured hUCB-MSCs on SIS gel and examined the angiogenic effect of conditioned medium on irradiated human umbilical vein endothelial cells (HUVECs) in vitro. RESULTS:hUCB-MSCs and SIS hydrogel composite treatment enhanced wound healing and angiogenesis in the wound site of mice. Conditioned medium from hUCB-MSCs cultured on SIS hydrogel promoted the chemotaxis of irradiated HUVECs more than their proliferation. The secretion of angiogenic growth factors hepatocyte growth factor, vascular endothelial growth factor-A and angiopoietin-1 from hUCB-MSCs was significantly increased by SIS hydrogel, with HGF being the predominant angiogenic factor of irradiated HUVECs. CONCLUSIONS:Our results suggest that the wound healing effect of hUCB-MSCs is enhanced by SIS hydrogel via a paracrine factor-mediated recruitment of vascular endothelial cells in a combined radiation-wound mouse model.
10.1016/j.jcyt.2017.06.007
In vivo release of bovine serum albumin from an injectable small intestinal submucosa gel.
Kang Kkot Nim,Kim Da Yeon,Yoon So Mi,Kwon Jin Seon,Seo Hyo Won,Kim E Sle,Lee Bong,Kim Jae Ho,Min Byoung Hyun,Lee Hai Bang,Kim Moon Suk
International journal of pharmaceutics
We aimed to develop a delivery system capable of maintaining a sustained release of protein drugs at specific sites using potentially biocompatible biomaterials. Here, we used bovine serum albumin (BSA) as a test protein to explore the potential utility of an injectable small intestine submucosa (SIS) as a depot for protein drugs. The prepared SIS powder was dispersed in PBS. The SIS suspension easily entrapped BSA in pharmaceutical formulations at room temperature. When this was suspension subcutaneously injected into rats, it gelled, forming an interconnecting three-dimensional network SIS structure to allow BSA to penetrate through it. The amount of BSA-FITC released from the SIS gel was determined in rat plasma and monitored by real-time in vivo molecular imaging. The data indicated the sustained release of BSA-FITC for 30 days in vivo. In addition, SIS gel provoked little inflammatory response. Collectively, our results show that the SIS gel described here could serve as a minimally invasive therapeutics depot with numerous benefits compared to other injectable biomaterials.
10.1016/j.ijpharm.2011.08.047
Production and Characterization of Composite Chitosan Hydrogel Containing Extracellular Matrix Particles for Tissue Engineering Applications.
Deng Xue-Qin,Chao Ning-Ning,Ding Wei,Qin Ting-Wu,Wang Wei,Zhang Yi,Luo Jing-Cong
Journal of biomedical nanotechnology
Chitosan-based hydrogels have been extensively used for tissue regeneration due to the excellent biocompatibility and biodegradability. For lack of endogenous extracellular biomacromolecules, its application is obviously limited. Because of robust biological activity, porcine small intestinal submucosa (SIS) has been considered as promising candidates to increase the bioactivity of hydrogels. Herein, a facile method for the fabrication of SIS powders (SISP)/chitosan chloride (CSCl)--glycerol phosphate (GP)-hydroxyethyl cellulose (HEC) hydrogel was developed. FTIR imaging results demonstrated that SISP and CSCl could be well mixed to form porous three-dimensional SISP/CSCl composite, which underwent sol-gel phage transition from solution to non-flowing hydrogel at 37 °C. Interestingly, the sustained release of VEGF and b-FGF within the composite hydrogel was determined and no initial burst release was observed. SISP/CSCl composite supported the survival and proliferation of NIH 3T3 cells and good biocompatibility in the SD rats subcutis up to 8 weeks. Furthermore, incorporated with SISP into CSCl delayed the degradation of SISP , as characterized by histological and High-Frequency Ultrasound (HFUS) measurement. Thus, all the findings suggested that the newlydeveloped injectable and thermosensitive SISP/CSCl composite was a promising and attractive candidate for soft tissue regeneration in the minimally-invasive way.
10.1166/jbn.2019.2713
Formation of three-dimensional tubular endothelial cell networks under defined serum-free cell culture conditions in human collagen hydrogels.
Andrée Birgit,Ichanti Houda,Kalies Stefan,Heisterkamp Alexander,Strauß Sarah,Vogt Peter-Maria,Haverich Axel,Hilfiker Andres
Scientific reports
Implementation of tubular endothelial cell networks is a prerequisite for 3D tissue engineering of constructs with clinically relevant size as nourishment of cells is challenged by the diffusion limit. In vitro generation of 3D networks is often achieved under conditions using serum containing cell culture medium and/or animal derived matrices. Here, 3D endothelial cell networks were generated by using human umbilical vein endothelial cells (HUVECs) in combination with human adipose tissue derived stromal cells (hASCs) employing human collagen I as hydrogel and decellularized porcine small intestinal submucosa as starter matrix. Matrigel/rat tail collagen I hydrogel was used as control. Resulting constructs were cultivated either in serum-free medium or in endothelial growth medium-2 serving as control. Endothelial cell networks were quantified, tested for lumen formation, and interaction of HUVECs and hASCs. Tube diameter was slightly larger in constructs containing human collagen I compared to Matrigel/rat tail collagen I constructs under serum-free conditions. All other network parameters were mostly similar. Thereby, the feasibility of generating 3D endothelial cell networks under serum-free culture conditions in human collagen I as hydrogel was demonstrated. In summary, the presented achievements pave the way for the generation of clinical applicable constructs.
10.1038/s41598-019-41985-6
Accelerating ESD-induced gastric ulcer healing using a pH-responsive polyurethane/small intestinal submucosa hydrogel delivered by endoscopic catheter.
Zhao Long-Mei,Gong Mei,Wang Rui,Yuan Qi-Juan,Zhang Yi,Pi Jin-Kui,Lv Xiu-He,Xie Yan,Xie Hui-Qi
Regenerative biomaterials
Endoscopic submucosal dissection (ESD) is the standard treatment for early-stage gastric cancer, but the large post-operative ulcers caused by ESD often lead to serious side effects. Post-ESD mucosal repair materials provide a new option for the treatment of post-ESD ulcers. In this study, we developed a polyurethane/small intestinal submucosa (PU/SIS) hydrogel and investigated its efficacy for accelerating ESD-induced ulcer healing in a canine model. PU/SIS hydrogel possessed great biocompatibility and distinctive pH-sensitive swelling properties and protected GES-1 cells from acid attack through forming a dense film in acidic conditions . Besides, PU/SIS gels present a strong bio-adhesion to gastric tissues under acidic conditions, thus ensuring the retention time of PU/SIS gels . In a canine model, PU/SIS hydrogel was easily delivered via endoscopy and adhered to the ulcer sites. PU/SIS hydrogel accelerated gastric ulcer healing at an early stage with more epithelium regeneration and slight inflammation. Our findings reveal PU/SIS hydrogel is a promising and attractive candidate for ESD-induced ulcer repair.
10.1093/rb/rbaa056
A biomimetic hierarchical small intestinal submucosa-chitosan sponge/chitosan hydrogel scaffold with a micro/nano structure for dural repair.
Wang Jingxi,Li Kun,Xu Junwei,Liu Meili,Li Ping,Li Xiaoming,Fan Yubo
Journal of materials chemistry. B
The dura mater is an essential barrier to protect the brain tissue and the dural defects caused by accidents can lead to serious complications. Various materials have been applied to dural repair, but it remains a challenge to perfectly match the structure and properties of the natural dura mater. Small intestinal submucosa has been developed for dural repair because of its excellent biocompatibility and biological activity, but its application is tremendously limited by the rapid degradation rate. Chitosan has also been broadly investigated in tissue repair, but the traditional chitosan hydrogels exhibit poor mechanical properties. A nanofiber chitosan hydrogel can be constructed based on an alkaline solvent, which is equipped with surprisingly high strength. Therefore, based on the bilayer structure of the natural dura mater, a biomimetic hierarchical small intestinal submucosa-chitosan sponge/chitosan hydrogel scaffold with a micro/nano structure was fabricated, which possessed a microporous structure in the upper sponge and a nanofiber structure in the lower hydrogel. The degradation rate was remarkably reduced compared with that of the small intestinal submucosa in the enzymatic degradation experiment . Meanwhile, the chitosan nanofibers brought high mechanical strength to the bilayer scaffold. Moreover, the hierarchical micro/nano structure and the active factors in the small intestinal submucosa have a fantastic effect on promoting the proliferation of fibroblasts and vascular endothelial cells. The bilayer scaffold showed good histocompatibility in the experiment of subcutaneous implantation in rats. Thus, the biomimetic hierarchical small intestinal submucosa-chitosan sponge/chitosan hydrogel scaffold with micro/nano structure simulates the structure of the natural dura mater and possesses properties with excellent performance, which has high practical value for dural repair.
10.1039/d1tb00948f
Immunosuppressed Miniswine as a Model for Testing Cell Therapy Success: Experience With Implants of Human Salivary Stem/Progenitor Cell Constructs.
Wu Danielle,Lombaert Isabelle M A,DeLeon Maximilien,Pradhan-Bhatt Swati,Witt Robert L,Harrington Daniel Anton,Trombetta Mark G,Passineau Michael J,Farach-Carson Mary C
Frontiers in molecular biosciences
An urgent need exists to develop large animal models for preclinical testing of new cell therapies designed to replace lost or damaged tissues. Patients receiving irradiation for treatment of head and neck cancers frequently develop xerostomia/dry mouth, a condition that could one day be treated by cell therapy to repopulate functional saliva-producing cells. Using immunosuppression protocols developed for patients receiving whole face transplants, we successfully used immunosuppressed miniswine as a suitable host animal to evaluate the long-term stability, biocompatibility, and fate of matrix-modified hyaluronate (HA) hydrogel/bioscaffold materials containing encapsulated salivary human stem/progenitor cells (hS/PCs). An initial biocompatibility test was conducted in parotids of untreated miniswine. Subsequent experiments using hS/PC-laden hydrogels were performed in animals, beginning an immunosuppression regimen on the day of surgery. Implant sites included the kidney capsule for viability testing and the parotid gland for biointegration time periods up to eight weeks. No transplant rejection was seen in any animal assessed by analysis of the tissues near the site of the implants. First-generation implants containing only cells in hydrogel proved difficult to handle in the surgical suite and were modified to adhere to a porcine small intestinal submucosa (SIS) membrane for improved handling and could be delivered through the da Vinci surgical system. Several different surgical techniques were assessed using the second-generation 3D-salivary tissue (3D-ST) for ease and stability both on the kidney capsule and in the capsule-less parotid gland. For the kidney, sliding the implant under the capsule membrane and quick stitching proved superior to other methods. For the parotid gland, creation of a tissue "pocket" for placement and immediate multilayer tissue closure were well tolerated with minimal tissue damage. Surgical clips were placed as fiduciary markers for tissue harvest. Some implant experiments were conducted with miniswine 90 days post-irradiation when salivation decreased significantly. Sufficient parotid tissue remained to allow implant placement, and animals tolerated immunosuppression. In all experiments, viability of implanted hS/PCs was high with clear signs of both vascular and nervous system integration in the parotid implants. We thus conclude that the immunosuppressed miniswine is a high-value emerging model for testing human implants prior to first-in-human trials.
10.3389/fmolb.2021.711602
Functional extracellular matrix hydrogel modified with MSC-derived small extracellular vesicles for chronic wound healing.
Cell proliferation
OBJECTIVES:Diabetic wound healing remains a global challenge in the clinic and in research. However, the current medical dressings are difficult to meet the demands. The primary goal of this study was to fabricate a functional hydrogel wound dressing that can provide an appropriate microenvironment and supplementation with growth factors to promote skin regeneration and functional restoration in diabetic wounds. MATERIALS AND METHODS:Small extracellular vesicles (sEVs) were bound to the porcine small intestinal submucosa-based hydrogel material through peptides (SC-Ps-sEVs) to increase the content and achieve a sustained release. NIH3T3 cell was used to evaluate the biocompatibility and the promoting proliferation, migration and adhesion abilities of the SC-Ps-sEVs. EA.hy926 cell was used to evaluate the stimulating angiogenesis of SC-Ps-sEVs. The diabetic wound model was used to investigate the function/role of SC-Ps-sEVs hydrogel in promoting wound healing. RESULTS:A functional hydrogel wound dressing with good mechanical properties, excellent biocompatibility and superior stimulating angiogenesis capacity was designed and facilely fabricated, which could effectively enable full-thickness skin wounds healing in diabetic rat model. CONCLUSIONS:This work led to the development of SIS, which shows an unprecedented combination of mechanical, biological and wound healing properties. This functional hydrogel wound dressing may find broad utility in the field of regenerative medicine and may be similarly useful in the treatment of wounds in epithelial tissues, such as the intestine, lung and liver.
10.1111/cpr.13196