Influence on the Osteogenic Activity of the Human Bone Marrow Mesenchymal Stem Cells Transfected by Liposome-Mediated Recombinant Plasmid pIRES-hBMP2-hVEGF165 In Vitro
Influence on the Osteogenic Activity of the Human Bone Marrow Mesenchymal Stem Cells Transfected by Liposome-Mediated Recombinant Plasmid pIRES-hBMP2-hVEGF165 In Vitro
复制标题
脂质体介导重组质粒pIRES-hBMP2-hVEGF165转染人骨髓间充质干细胞体外成骨活性的影响
DOI：
10.1097/sap.0b013e3181b4bc5d
复制
发表时间：
2010-07-01
影响因子：
1.5
通讯作者：
Guo Li
中科院分区：
文献类型：
--
作者：
Wu Guo-ping;He Xiao-chuan;Guo Li
骨髓间充质干细胞 (BMSC) 是在组织工程策略中用作种子细胞的一个有吸引力的选择。骨形态发生蛋白 (BMP) 参与各种组织类型的形成，包括骨、软骨、肌腱和韧带。血管内皮生长因子(VEGF)是一种很有前景的诱导血管生成的试剂。构建了人骨形态发生蛋白2(hBMP-2)和血管内皮生长因子165(hVEGF165)的共表达载体。将载体转染到从健康成人骨髓中分离的增殖的 BMSC 中。采用Western-blot检测BMSCs hBMP-2和hVEGF165基因的表达情况，并通过RT-PCR分析骨钙素mRNA表达情况测定BMSCs碱性磷酸酶活性水平。免疫组化染色测定I型胶原水平。重组质粒pIRES-hBMP-2-VEGF165转染BMSCs后，可分泌高水平的BMP-2和hVEGF165。与对照组相比，转染的BMSCs中I型胶原的产生、碱性磷酸酶的活性和骨钙素mRNA的表达也显着提高。外源hBMP-2和hVEGF165基因可以在转染的BMSCs中组成型表达，并且含有基因组合的BMSCs的谱系定向分化能力 BMP-2 和 VEGF165 增强。
Bone marrow mesenchymal stem cells (BMSCs) is an attractive option for use as seed cells in tissue engineering strategies. Bone morphogenetic proteins (BMPs) to be involved in the formation of various tissue types including bone, cartilage, tendon, and ligament. Vascular endothelial growth factor (VEGF) is a promising reagent for inducing angiogenesis.Constructed a coexpressing vector of human bone morphogenetic protein 2 (hBMP-2) and vascular endothelial growth factor 165 (hVEGF165). The vectors were transfected into proliferated BMSCs isolated from healthy adult bone marrow. The expression of hBMP-2 and hVEGF165 genes of BMSCs were assayed by Western-blot, and the level of alkaline phosphatase activities of BMSCs was determined by RT-PCR analysis of osteocalcin mRNA expression. The levels of collagen I by immunohistochemical staining were also determined.BMSCs transfected with reconstructed plasmid pIRES-hBMP-2-VEGF165 could secret a high level of BMP-2 and hVEGF165. The production of type I collagen, the activity of alkaline phosphatase, and the expression of osteocalcin mRNA were also significantly improved in the transfected BMSCs, compared with the control group.The exogenous hBMP-2 and hVEGF165 genes can be expressed constitutively in the transfected BMSCs, and the lineage-committed differentiation abilities of BMSCs containing combination of genes BMP-2 and VEGF165 are enhanced.