骨形态发生蛋白2和血管内皮生长因子165双基因转染骨髓基质干细胞的异位诱导成骨能力

发布时间：2018-05-11 02:37

本文选题：干细胞 + 骨髓　；参考：《中国组织工程研究》2017年09期

【摘要】：背景:应用骨形态发生蛋白2(BMP2)和血管内皮生长因子165(VEGF165)双基因转染骨髓基质干细胞诱导成骨,为组织工程骨的研究提供实验基础。目的:探讨BMP2和VEGF165双基因转染大鼠骨髓基质干细胞的异位诱导成骨能力。方法:4周龄SD大鼠4只,取股骨、胫骨骨髓,采用全骨髓贴壁法分离培养骨髓基质干细胞,取第3代骨髓基质干细胞分为5组,分别为未转染组、空载质粒组、BMP2单基因转染组、VEGF165单基因转染组、BMP2和VEGF165双基因共转染组,转染后48 h通过Western Blot检测BMP2和VEGF165蛋白表达变化,转染后7d检测各组细胞的碱性磷酸酶活性。结果与结论:(1)BMP2和VEGF165双基因共转染组和BMP2单基因转染组中有大量的BMP2分泌。BMP2和VEGF165双基因共转染组和VEGF165单基因转染组中有大量的VEGF165分泌。双基因共转染组中BMP2和VEGF165蛋白水平与单基因转染组比较,差异有显著性意义(P0.05);(2)BMP2和VEGF165双基因共转染组中碱性磷酸酶活性最高,其次是BMP2单基因转染组,而VEGF165单基因转染组明显不如前两组,但是略高于空质粒转染组和未转染组。统计分析表明双基因共转染组与单基因转染组比较,差异有显著性意义(P0.05);(3)结果表明,BMP2和VEGF165双基因转染促进骨髓基质干细胞异位成骨的能力更强。
[Abstract]:Background: bone morphogenetic protein 2 (BMP2) and vascular endothelial growth factor 165 (VEGF165) were transfected into bone marrow stromal cells to induce osteogenesis. Aim: to investigate the ability of ectopic osteogenesis of rat bone marrow stromal stem cells transfected with BMP2 and VEGF165. Methods Bone marrow stromal cells (BMSCs) were isolated from bone marrow of femur and tibia of 4 week old Sprague-Dawley rats. Bone marrow stromal cells (BMSCs) of the third generation were divided into 5 groups: untransfected group. The expression of BMP2 and VEGF165 protein was detected by Western Blot 48 h after transfection, and alkaline phosphatase activity was detected 7 days after transfection. Results and conclusion there was a large amount of VEGF165 secretion in the two gene cotransfection groups: BMP2 and VEGF165 and BMP2 single gene transfection group. There was a large amount of VEGF165 secretion in the BMP2 and VEGF165 co-transfection groups and VEGF165 single gene transfection group. The levels of BMP2 and VEGF165 protein in the two gene cotransfection group were significantly higher than those in the single gene transfection group, and the activity of alkaline phosphatase was the highest in the two gene cotransfection groups, followed by the BMP2 single gene transfection group. However, the single gene transfection group of VEGF165 was significantly lower than the former two groups, but slightly higher than the empty plasmid transfection group and the untransfected group. Statistical analysis showed that there was a significant difference between the two gene cotransfection group and the single gene transfection group. The results showed that the double gene transfection of BMP2 and VEGF165 could promote the heterotopic osteogenesis of bone marrow stromal stem cells (BMSCs).
【作者单位】：西南医科大学附属口腔医院口腔颌面外科;
【基金】：四川省科技厅应用基础研究资助项目(2008jy0014)~~
【分类号】：R318.08

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