人骨形态发生蛋白-2重组慢病毒载体转染羊骨髓间充质干细胞及其成骨调节作用

目的探讨人骨形态发生蛋白-2（bone morphogenetic protein-2，BMP-2）基因转染对体外培养的骨髓间充质干细胞（bone marrow mesenchymal stem cells，BMSCs）增殖和骨向分化的影响。方法构建携带人BMP2基因的慢病毒载体，转染诱导羊第4代BMSCs为转染组，选择未转染的BMSCs为对照组。采用Western blot检测2组细胞BMP-2与成骨相关蛋白表达情况，采用实时定量PCR法检测细胞I型胶原、骨钙素、骨桥蛋白mRNA相对表达量的变化情况。结果转染组骨钙素、骨桥蛋白mRNA及蛋白表达量较对照组增高，差异有统计学意义（P〈0．05），2组I型胶原mRNA及蛋白表达量差异无统计学意义（P〉0．05）。结论BMP-2基因转染可增强羊BMSCs增殖和骨向分化的能力，可作为组织工程骨的种子细胞。

Recombinant lentiviral vector of human bone morphogenetic protein-2 transfected sheep bone marrow-derived mesenchymal stem cells and its osteogenic effect

Objective To study the effect of human bone morphogenetic protein-2 （BMP-2） transfection on the proliferation and osteogenic differentiation of in vitro bone marrow mesenchymal stem cells （BMSCs）. Methods Lentiviral vectors carrying BMP-2 were constructed to transfect BMSCs in sheep of the fourth generation. The non- transfected BMSCs in BMSCs in sheep were as controls （control group）. Western blot was used to detect the expressions of BMP-2 and osteoblast-related protein, and real-time quantitative PCR was used to detect the mRNA levels of collagen- I, osteocalcin and osteopontin. Results The expressions of osteocalcin and osteopontin mRNA and protein were higher in transfection group than those in control group （P〈0.05）, and there was no significant difference in the expressions of collagen-I mRNA and protein between two groups （P〉0.05）. Conclusions BMP-2 transfection can enhance the ability of BMSCs proliferation and osteogenic differentiation, and the transfected cells are suitable for seed cells for bone tissue engineering.