目的 构建细胞外基质( ECM )-多糖/蛋白质多孔复合支架材料,观察骨髓间充质干细胞(BMSCs)在其上的生长、增殖和成软骨分化的行为,探讨该材料用于体外构建组织工程软骨的可行性.方法采用仿生学方法,将卡拉胶、明胶、壳聚糖按一定比例制成ECM -多糖/蛋白质多孔复合支架材料,扫描电镜观察其形态. MTT法检测软骨细胞在材料上的增殖行为.将BMSCs接种于复合支架材料上,并设软骨细胞诱导液组为对照组,观察细胞的生长情况,免疫组化检测Ⅱ型胶原蛋白的表达.结果 复合材料呈多孔隙海绵状,扫描电镜观察材料呈三维多孔的网状结构.软骨细胞在支架材料上增殖明显,MTT结果显示实验组和对照组各点OD值之间差异无统计学意义(P＞0.05).支架材料具备诱导BMSCs向成软骨细胞转化的功能,两组细胞都可以检测到Ⅱ型胶原蛋白的表达.结论 ECM-多糖/蛋白质多孔复合材料具有良好的生物相容性,在不添加诱导剂的条件下,可以诱导BMSCs向软骨细胞分化.%Objective To construct the extracellular matrix ( ECM)-polysaccharide/protein porous composite scaffold; thus to observe the growth, proliferation and chondrogenic differentiation behavior of bone marrow mesenchymal stem cells (BMSCs) on the scaffold, and the application in engineering cartilage in vitro .Methods Carrageenan, gelati-n and chitosan were used for bionic construction of ECM -polysaccharide/protein porous composite scaffolds .The mor-phology of the scaffolds was observed by SEM .The proliferation of chondrocytes on the scaffolds was detected by MTT .In vitro cartilage tissue formation of BMSCs on scaffolds were shown by type Ⅱcollagen immunohistochemistry staining , using OB as control.Results The scaffolds were porous with three -dimension porous network structure according to SEM ob-servation.The proliferation of chondrocytes on scaffolds was obvious , and there was no significant difference between the experimental group and the control group (P＞0.05).The expression of type Ⅱ collagen was detected in both groups . Conclusion The scaffolds have good biocompatibility and can induce BMSCs differentiate into chondrocytes without OB .
展开▼
