Ad-hBMP-2与Ad-hTGF-β1基因共转染骨髓间充质干细胞后的体外成骨诱导  

In vitro osteogenic induction of bone marrow mesenchymal stem cells after co-transfection with Ad-h BMP-2 and Ad-h TGF-β1

作　　者：陈剑平[1] 邱俊钦[1] 曾昭勋[1] 陈宗雄[1] 尹承慧[1] 

机构地区：[1]解放军南京军区福州总医院骨一科,福建省福州市350025

出　　处：《中国组织工程研究》2014年第50期8037-8041,共5页Chinese Journal of Tissue Engineering Research

基　　金：福建省青年科技人才创新项目(2005J075)~~

摘　　要：背景:重组DNA技术的进展为骨组织工程研究的发展提供巨大动力,将其应用于组织工程骨构建可以显著提高骨修复的质量和效率。目的:拟将Ad-h BMP-2联合Ad-h TGF-β1转染骨髓间充质干细胞体外成骨诱导,观察二者在成骨过程中的相互关系。方法:以腺病毒Ad Max为基因转移载体,将携带转化生长因子β1和骨形态发生蛋白2基因的高滴度腺病毒感染SD大鼠骨髓间充质干细胞,利用外源性基因编码的生长因子诱导其表达,通过RT-PCR、ELISA、MTT实验及碱性磷酸酶水平检测等方法鉴定。结果与结论:腺病毒感染骨髓间充质干细胞后RT-PCR可检测出外源基因的表达,MTT实验和碱性磷酸酶活性检测双基因共转染的骨髓间充质干细胞增殖能力最强,碱性磷酸酶活性最高。结果表明联合应用Ad-h TGF-β1和Ad-h BMP-2转染可明显促进骨髓间充质干细胞的成骨化,效果优于单用一种生长因子。BACKGROUND： Advances in recombinant DNA technology provide a huge impetus to the development of bone tissue engineering research, which used for tissue-engineered bone construction can significantly improve the quality and efficiency of bone repair. OBJECTIVE： To evaluate the in vitro osteogenic induction of Ad-hBMP-2 and Ad-hTGF-131 co-transfected bone marrow mesenchymal stem cells and to elucidate their relationship in bone formation. METHODS： Bone marrow mesenchymal stem cells from Sprague-Dawley rats were infected by high-titer recombinant adenoviruses carrying transforming growth factor-βl and bone morphogenetic protein-2 genes, and then transfected cells were induced by exogenous genes detected with RT-PCR, ELISA, MTT and alkaline phosphatase test. RESULTS AND CONCLUSION： Adenovirus-infected bone marrow mesenchymal stem cells could express exogenous genes detected by RT-PCR. MTT test and alkaline phosphatase test showed the proliferation ability of co-transfected bone marrow mesenchymal stem cells was the strongest and the alkaline phosphatase activity was the highest. It has been observed that bone marrow mesenchymal stem cells co-transfected with Ad-hBMP-2 and Ad-hTGF-~I can be induced to form bone tissue＇cooperatively and synergistically. Subject headings： bone marrow; mesenchymal stern cells; transforming growth factor beta1; bone morphogenetic proteins Funding： the Scientific Young Talent Innovative Project of Fujian Province, No. 2005J075

关 键 词：骨髓 间质干细胞 转化生长因子β1 骨形态发生蛋白质类 干细胞 骨髓干细胞 骨形态发生蛋白2 腺病毒载体 

分 类 号：R394.2[医药卫生—医学遗传学]