成骨细胞特异性钙黏蛋白基因转染对人骨髓基质干细胞成骨分化影响的研究

目的 观测成骨细胞特异性钙黏蛋白(Cad-Ⅱ)基因转染对人骨髓基质干细胞(hBMSCs)成骨分化的影响. 方法把脂质体介导的Cad-Ⅱ cDNA转染体外分离培养的hBMSCs,检测Cad-Ⅱ蛋白表达变化,并对比观测转染组和单纯成骨诱导组在培养3,7、14、21 d时,hBMSCs碱性磷酸酶(ALP)和骨钙索的表达变化. 结果 转染组和成骨诱导组3 d后开始有ALP的阳性染色,呈棕黑色,7 d开始有骨钙索的阳性染色并随培养时间延长逐渐增多,但各时间点转染组ALP浓度(P=0.008)和骨钙素染色阳性数(P=0.023)均显著高于单纯成骨诱导组.转染组和成骨诱导组从14 d后开始有红色的刚件染色矿化结节出现,结节数目随时间推移而增多. 结论 Cad-Ⅱ基因转染可促进hBMSCs向成骨细胞的分化.

Effect of OB-cadherin gene transfection on osteogeule differentiation of human mesenchymal stem cells in vitro

Objective To study the effects of Cad-Ⅱgene on osteogenic differentiation of human mesenchymal stem cells in vitro. Methods The human marrow mesenchymal stem cells (hBMSCs) were isolated and cultured in vitro before they were divided into 2 groups. In the transfeetion group, the hBMSCs were transfeeted with Cad- Ⅱgene;in the simple osteogenic inducement group, they were cultured in the condition medium. Then the expressions of Cad- Ⅱ protein were determined and the alkaline phosphatase (ALP) activities and the expressions of ostcocalein were measured at 3, 7, 14 and 21 days for comparison between the 2 groups. Results The ALP activity and positive expression of osteoealcin were regulated significantly higher in the transfeetion group than in the simple osteogenic inducement group at different times (P <0.05). The mineralized nodes began to appear at 14 days in the 2 groups and increased with time. Conclusion Cad-Ⅱgene transfeetion can promote differentiation of hBMSCs into the osteoblasts.