Due to the unique microstructures and components of extracellular matrix(ECM),decellularized scaffolds had been used widely in clinical.The reaction of the host toward decellularized scaffolds depends on their biocomp...Due to the unique microstructures and components of extracellular matrix(ECM),decellularized scaffolds had been used widely in clinical.The reaction of the host toward decellularized scaffolds depends on their biocompatibility,which should be satisfied before applied in clinical.The aim of this study is to develop a decellularized xenograft material with good biocompatibility for further bone repair,in an effective and gentle method.The existing chemical and physical decellularization techniques including ethylene diamine tetraacetic acid(EDTA),sodium dodecyl sulfate(SDS)and supercritical carbon dioxide(SC-CO2)were combined and modified to decellularize bovine cancellous bone(CB).After decellularization,almost 100%of A-Gal epitopes were removed,the combination of collagen,calcium and phosphate was reserved.The direct and indirect contact with macrophages was used to evaluate the cytotoxicity and immunological response of the materials.Mesenchymal stem cells(MSCs)were used in the in vitro cells’proliferation assay.The decellularized CB was proved has no cytotoxicity(grade 1)and no immunological response(NO,IL-2,IL-6 and TNF-α secretion inhibited),and could support MSCs proliferated continuedly.These results were similar to that of commercial decellularized human bone.This study suggests the potential of using this kind of combine decellularization process to fabricate heterogeneous ECM scaffolds for clinical application.展开更多
目的:体外构建组织工程软骨,筛选更为适合组织工程软骨构建的支架材料。方法:体外获取SD大鼠肋软骨细胞。采用第一代软骨细胞作为种子细胞,接种于壳聚糖/明胶和BMG/生物蛋白胶支架,体外培养的不同时间对其进行HE、甲苯胺蓝染色、Mas s o...目的:体外构建组织工程软骨,筛选更为适合组织工程软骨构建的支架材料。方法:体外获取SD大鼠肋软骨细胞。采用第一代软骨细胞作为种子细胞,接种于壳聚糖/明胶和BMG/生物蛋白胶支架,体外培养的不同时间对其进行HE、甲苯胺蓝染色、Mas s on染色、免疫学检测、扫描电镜观察。结果:在培养2周时,BMG/生物蛋白胶各种染色结果显示软骨细胞在其表面以及内部分布均匀,蛋白多糖和Ⅱ型胶原染色阳性;壳聚糖/明胶表面细胞稍多于前者,但内部细胞数量极少且分布不均,染色结果不如前者明显。随着培养时间的延长各种检测均显示有大量的软骨细胞特异性的蛋白多糖和Ⅱ型胶原的表达,壳聚糖/明胶凸显出明显的优势。结论:体外成功构建组织工程软骨,软骨细胞在BMG/生物蛋白胶上的生长、增殖和分泌基质情况优于壳聚糖/明胶支架。展开更多
基金supported by National Natural Science Foundation of China(51502094)Special Fund Project for Guangdong Academy of Sciences to Build First-Class Research Institutions in China(2020GDASYL-20200103038)+4 种基金National Key R&D Plans(2018YFC1105902)China Postdoctoral Science Foundation(2017M612657)Guangdong Province Science and Technology Projects(2013B021800137)Pearl River Nova Program of Guangzhou(201610010168)Guangdong Province Medical Research Foundation(A2016060).
文摘Due to the unique microstructures and components of extracellular matrix(ECM),decellularized scaffolds had been used widely in clinical.The reaction of the host toward decellularized scaffolds depends on their biocompatibility,which should be satisfied before applied in clinical.The aim of this study is to develop a decellularized xenograft material with good biocompatibility for further bone repair,in an effective and gentle method.The existing chemical and physical decellularization techniques including ethylene diamine tetraacetic acid(EDTA),sodium dodecyl sulfate(SDS)and supercritical carbon dioxide(SC-CO2)were combined and modified to decellularize bovine cancellous bone(CB).After decellularization,almost 100%of A-Gal epitopes were removed,the combination of collagen,calcium and phosphate was reserved.The direct and indirect contact with macrophages was used to evaluate the cytotoxicity and immunological response of the materials.Mesenchymal stem cells(MSCs)were used in the in vitro cells’proliferation assay.The decellularized CB was proved has no cytotoxicity(grade 1)and no immunological response(NO,IL-2,IL-6 and TNF-α secretion inhibited),and could support MSCs proliferated continuedly.These results were similar to that of commercial decellularized human bone.This study suggests the potential of using this kind of combine decellularization process to fabricate heterogeneous ECM scaffolds for clinical application.
文摘目的:体外构建组织工程软骨,筛选更为适合组织工程软骨构建的支架材料。方法:体外获取SD大鼠肋软骨细胞。采用第一代软骨细胞作为种子细胞,接种于壳聚糖/明胶和BMG/生物蛋白胶支架,体外培养的不同时间对其进行HE、甲苯胺蓝染色、Mas s on染色、免疫学检测、扫描电镜观察。结果:在培养2周时,BMG/生物蛋白胶各种染色结果显示软骨细胞在其表面以及内部分布均匀,蛋白多糖和Ⅱ型胶原染色阳性;壳聚糖/明胶表面细胞稍多于前者,但内部细胞数量极少且分布不均,染色结果不如前者明显。随着培养时间的延长各种检测均显示有大量的软骨细胞特异性的蛋白多糖和Ⅱ型胶原的表达,壳聚糖/明胶凸显出明显的优势。结论:体外成功构建组织工程软骨,软骨细胞在BMG/生物蛋白胶上的生长、增殖和分泌基质情况优于壳聚糖/明胶支架。
