定向诱导骨髓间充质干细胞-藻酸盐复合物构建软骨组织

目的研究骨髓间充质干细胞(M SC s)经向软骨细胞定向诱导后,与藻酸盐复合,构建组织工程软骨的可能性。方法取大鼠股骨骨髓,经梯度离心法和贴壁法分离M SC s,经鉴定后用含转化生长因子1β10 ng/m l、地塞米松1-0 7m o l/L、转铁蛋白3 m g/m l、胰岛素2 m g/m l的诱导因子培养基对M SC s进行诱导,14 d后免疫组化检测Ⅱ型胶原分泌情况,将诱导后和未经诱导的M SC s以1×106/m l细胞密度与藻酸钙复合接种于裸鼠,分别于4周、8周后行组织学检测。结果经诱导的M SC sⅡ型胶原免疫组化阳性,与藻酸盐复合4周,HE染色可见软骨样细胞、软骨陷窝形成,阿辛蓝染色和M asson染色可见新生的软骨细胞和细胞外胶原,8周后软骨细胞明显增多,软骨细胞分泌胶原丰富,而未经诱导的M SC s无明显软骨细胞生成。结论M SC s经定向诱导后,可与藻酸钙复合在体内形成软骨样组织,可能发展为临床修补软骨缺损的一条有效途径。

Experimental Study of Cartilage Reconstruction Using Directed Inducing MSCs-Alginate Complex

OBJECTIVE: To evaluate the practicability of cartilage reconstruction using directed inducing bone marrow stem cells(MSCs)- alginate complex. METHODS: The MSCs were seperated by gradient centrifugation on Percoll, then were induced into chon-drogenic differentiation. After 14 days,immunohistochemical techique was applied to detect the expression of collagen type II. Then, the MSCs and calcium alginate were mixed to form a complex and were planted under the skin of nude mice. Histochemical or immuno-histochemical tests were made at 4 weeks and 8 weeks later. RESULTS: The MSCs showed the positive result of collagen type II. There were chondrcytes and cartilage lacuna at 4 weeks later, and chondrocytes and the collagen secreted by them became more at 8 weeks later. But, the MSCs without directed inducing did not show the character of the chondrocyte. CONCLUSION: The way of using directed inducing MSCs-alginate complex can form cartilage; it is possible to repair the defect of cartilage in clinical practice.