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目的比较纤维蛋白凝胶与几丁质对骨髓间充质干细胞(BMSCs)向软骨细胞分化的影响,探讨三维支架与软骨组织工程种子细胞BMSCs分化的关系。方法BMSCs与几丁质、纤维蛋白凝胶形成复合物,分别体外培养及植入大鼠关节软骨缺损部位。体外培养14d后,进行HE染色、甲苯胺蓝及Ⅱ型胶原免疫组织化学染色;体内培养2周、4周、6周后,对移植物进行形态学观察,表达软骨特异蛋白分析及BMSCs体内示踪。统计学分析BMSCs向软骨分化情况。结果体外培养部分,BMSCs-纤维蛋白凝胶组和BMSCs-几丁质组的Ⅱ型胶原免疫组织化学染色阳性率与对照组无显著差异;体内移植部分,BMSCs-纤维蛋白凝胶组的甲苯胺蓝染色与Ⅱ型胶原免疫组织化学染色积分吸光度(IA)变化率与对照组有显著差异,其他组别软骨分化与对照组无显著差异。结论在体外纤维蛋白凝胶或几丁质诱导BMSCs向软骨细胞分化的作用很弱,在体内BMSCs-纤维蛋白凝胶可促进BMSCs分化成类软骨细胞。
Objective To compare the effect of fibrin gel and chitin on the differentiation of bone marrow mesenchymal stem cells (BMSCs) into chondrocytes and to explore the relationship between the three-dimensional scaffold and the differentiation of BMSCs in cartilage tissue engineering seed cells. Methods BMSCs formed complexes with chitin and fibrin gel, and cultured in vitro and implanted into the defect site of articular cartilage in rats. After cultured in vitro for 14 days, HE staining, toluidine blue staining and type Ⅱ collagen immunohistochemical staining were performed. After 2 weeks, 4 weeks and 6 weeks of in vivo culture, the grafts were observed for morphological observation and cartilage-specific protein analysis and BMSCs expression in vivo trace. Statistical analysis of BMSCs to cartilage differentiation. Results The positive rate of type Ⅱ collagen immunohistochemistry in BMSCs-fibrin gel group and BMSCs-chitin group was not significantly different from that in control group. In vivo transplanted tissue, the toluidine in BMSCs-fibrin gel group Blue staining and type Ⅱ collagen immunohistochemical staining integral absorbance (IA) rate of change was significantly different from the control group, other groups of cartilage differentiation and the control group no significant difference. CONCLUSION: BMSCs-fibrin gel can promote the differentiation of BMSCs into chondrocytes induced by fibrin gel or chitin in vitro.