骨形态发生蛋白2和7共转染骨髓间充质干细胞的成骨分化  

Bone morphogenetic protein 2 co-transfected with bone morphogenetic protein 7 for osteogenic differentiation of bone marrow mesenchymal stem cells

作　　者：陈剑[1,2] 袁文[3] 宋滇文[3] 胡凯猛[4] 范立星[4] 刘厚奇[4] 

机构地区：[1]浙江大学医学院附属邵逸夫医院 骨科 [2]浙江大学医学院附属邵逸夫医院 临床研究所,浙江省杭州市310016 [3]解放军第二军医大学附属长征医院骨科,上海市200003 [4]解放军第二军医大学组织胚胎学教研室,上海市200433

出　　处：《中国组织工程研究》2012年第49期9139-9145,共7页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金项目(30970735),转基因骨髓间充质干细胞与自体血小板凝胶生物支架构建组织工程骨的实验研究~~

摘　　要：背景:骨形态发生蛋白最主要的作用是诱导骨形成,但因提取困难、代谢速度快、难以准确控制其使用浓度、价格昂贵等限制了其在体外和体内相关研究中的应用。目的:构建含特异细胞生长因子基因骨形态发生蛋白2,7的腺病毒载体,观察骨形态发生蛋白2和骨形态发生蛋白7基因共转染对兔骨髓间充质干细胞成骨分化的促进作用。方法:将全骨髓贴壁法分离得到兔骨髓间充质干细胞传至第3代,分为5组,空白组和常规诱导组分别以常规培养基和成骨诱导分化培养基培养;骨形态发生蛋白2,7腺病毒转染组:分别单独以骨形态发生蛋白2、骨形态发生蛋白7腺病毒进行转染;联合转染组:以2种骨形态发生蛋白腺病毒联合转染。结果与结论:转染第7天,联合转染组成骨相关基因Runx-2、Osx、Ⅰ型胶原和碱性磷酸酶mRNA表达均较其他各组增高(P<0.05);骨形态发生蛋白2,7腺病毒转染组上述指标较空白组和常规诱导组增高(P<0.05)。转染第14天,联合转染组4个指标均较其他各组明显增高(P<0.05);同时,4个指标表达均高于第7天(P<0.05)。病毒转染后第7天,联合转染组Ⅰ型胶原和骨钙素蛋白表达均较其他组明显增高(P<0.05)。提示对骨髓间充质干细胞进行骨形态发生蛋白2腺病毒和骨形态发生蛋白7腺病毒共转染后,较单基因转染和成骨诱导液诱导更能促进成骨相关基因和蛋白的表达。BACKGROUND： The most important role of bone morphogenetic protein is the induction of bone formation some factors including difficulty in sample extract, rapid metabolic rate, difficulty in accurate concentration control and high cost, limit the application for in vitro and in vivo studies. OBJECTIVE： To establish the adenoviral vectors containing bone morphogenetic protein 2 and bone morphogenetic protein 7 in order to observe the promotion effect of co-transfection of bone morphogenetic protein 2 and bone morphogenetic protein METHODS： The rabbit bone marrow mesenchymal stem cells separated by the whole bone marrow adherence method were passaged to the third generation, and divided into five groups： the control group and the conventional induction group were cultured with conventional culture medium and osteo-induction culture medium; bone morphogenetic protein 2 group and bone morphogenetic protein 7 group were transfected with bone morphogenetic protein 2 adenovirus and bone morphogenetic protein 7 adenovirus; the combination group was co-transfected with bone morphogenetic protein 2 adenovirus and bone morphogenetic protein 7 adenovirus. RESULTS AND CONCLUSION： At 7 days after transfection, the expression levels of Runx-2, Osx, alkaline phosphatase and Collagen Ⅰ mRNA in the combination group were higher than those in the other four groups （P 〈 0.05）; the indicators above in the bone morphogenetic protein 2 group and bone morphogenetic protein 7 group were higher than those in control arouo and the conventional induction group （P 〈 0,05）. At 14 days after transfection, the expression of Runx-2, Osx,alkaline phosphatase and collagen Ⅰ mRNA in the combination group were higher than those in the other four groups （P 〈 0.05）; and the indicators at 14 days after transfection were higher than those at 7 days after trasnfection （P 〈 0.05）. At 7 days after transfection, the expression levels of collagen Ⅰand osteocalcin protein in the combination group were higher than those i

关 键 词：骨髓间充质干细胞 干细胞 腺病毒 骨形态发生蛋白2 骨形态发生蛋白7 Ⅰ型胶原 共转染 成骨诱导 成骨活性 成骨蛋白 组织工程 

分 类 号：R394.2[医药卫生—医学遗传学]