纤维蛋白凝胶与几丁质对骨髓间充质干细胞向软骨细胞分化的影响  

Effect of fibrinogen gel and chitin on BMSCs to chondrocytes differentiation

作　　者：李冬梅[1] 金连弘[2] 张宝东[2] 杨水友[1] 李继承[1] 

机构地区：[1]浙江大学医学院基础医学院,杭州310058 [2]哈尔滨医科大学组织学胚胎学教研室,哈尔滨150086

出　　处：《解剖学报》2009年第6期909-913,共5页Acta Anatomica Sinica

基　　金：国家基础科学人才培养资助项目(J0730856;J0830833);浙江省教育厅资助项目(Y200804201)

摘　　要：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 We compare tile effects of fibrinogen gel and chitin on BMSCs to chondrocytes differentiation in order to explore the relationship between three-dimensional scaffold and cartilage lissue engineering seed cells BMSCs differentiation. Methods BMSCs together with chitin and fibrin gel complexes were cultured in ritro and implanted into rat＇ s articular cartilage defect location. After 14 days in vitro culture, HE staining, toluidine blue staining and type Ⅱ collagen immunohistochemical staining were performed; After transplantation in vivo, for 2 weeks, 4 weeks and 6 weeks, morpholog7 observations, expression of cartilage-specific protein analysis and BMSCs in viro tracer method were performed. We analyzed the differentiation of BMSCs into chondrocytes by statistical methods. Results The comparison of positive cell rate of type II collagen immunohistochemical staining in BMSCs-fibrin gel group and BMSCs-chitin group which were cultured in vitro, showed no significant difference with control group. Integrated absorbance（IA ） change rate of toluidine blue staining and type Ⅱ collagen immunohistochemical staining in BMSCs-fibfin gel group which was cultured in vivo, showed significantly different with other groups and control group. Conclusion The results showed that in vitro fibrin gel or chitin has very weak induction of BMSCs to cartilage differentiation, while in vivo BMSCs-fibrin gel can facilitrate BMSCs to differentiate into chondrocyte-like cells.

关 键 词：骨髓间充质干细胞 软骨细胞 纤维蛋白凝胶 几丁质 免疫组织化学 大鼠 

分 类 号：R322.7[医药卫生—人体解剖和组织胚胎学]