Combination of BMP9 and VEGF induces osteogenic differentiation of C3H10T1/2 mouse mesenchymal stem cells in vitro

He Yin; He Yun

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Combination of BMP9 and VEGF induces osteogenic differentiation of C3H10T1/2 mouse mesenchymal stem cells in vitro

更新时间：2023-12-18

- Combination of BMP9 and VEGF induces osteogenic differentiation of C3H10T1/2 mouse mesenchymal stem cells in vitro
- Journal of Chongqing Medical University Vol. 40, Issue 2, Pages: 197-202(2015)
- 作者机构：
  
  1. 重庆市第九人民医院儿科,重庆,400700
  2. 重庆医科大学附属儿童医院儿科研究所小儿外科,重庆,400014
- 作者简介：
- 基金信息：
- DOI：
  CLC：
- Published：2015，
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He Yin, He Yun. Combination of BMP9 and VEGF induces osteogenic differentiation of C3H10T1/2 mouse mesenchymal stem cells in vitro. [J]. Journal of Chongqing Medical University 40(2):197-202(2015)
DOI：

He Yin, He Yun. Combination of BMP9 and VEGF induces osteogenic differentiation of C3H10T1/2 mouse mesenchymal stem cells in vitro. [J]. Journal of Chongqing Medical University 40(2):197-202(2015) DOI：

摘要

目的：探讨重组腺病毒介导的骨形态发生蛋白9（bone morphogenetic protein 9，BMP9）和血管内皮细胞生长因子（vas－cular endothelial growth factor，VEGF）共感染对骨髓间充质干细胞（bone marrow mesenchymal stem cells，BMSCs）成骨分化的影响。方法：将重组腺病毒介导的BMP9和VEGF分5组感染小鼠间充质干细胞C3H10T1/2，Ad-BMP9组、Ad-VEGF组、Ad-VEGF+Ad-BMP9组、空载体腺病毒组和空白组，感染7 d后，real-time PCR检测BMP9和VEGF的mRNA水平，real-time PCR检测成骨指标骨钙蛋白（osteocalcin，OC）、骨保护素（osteoprotegerin，OPG）、骨桥蛋白（osteopontin，OPN）的mRNA水平，Western blot检测OPN蛋白的表达，各组细胞行碱性磷酸酶（alkaline phosphatase，ALP）活性的定量检测、ALP染色和钙盐沉积的检测。结果：BMP9和VEGF在Ad-VEGF+Ad-BMP9组高效共表达，OC、OPG、OPN mRNA水平和OPN蛋白水平在Ad-BMP9组和Ad-VEGF+Ad-BMP9组的表达明显高于其他各组（P﹤0.05），且在Ad-VEGF+Ad-BMP9组表达最高（P﹤0.05）。Ad-VEGF+Ad-BMP9组的ALP活性、ALP染色和钙盐沉积明显高于其他各组（P﹤0.05）。结论：联合VEGF可明显增强BMP9诱导C3H10T1/2细胞成骨分化能力。

Abstract

Objective：To investigate the effects of recombinant adenovirus-mediated co-infection of bone morphogenetic protein 9（BMP9） and vascular endothelial growth factor（VEGF） gene on osteogenic differentiation of bone marrow mesenchymal stem cells（BMSCs）. Methods：C3H10T1/2 cells were infected by recombinant adenovirus-mediated of BMP9 and VEGF as follow five groups：Ad-BMP9 group，Ad-VEGF group，Ad-VEGF+Ad-BMP9 group，Ad-null group and control group. After infecting for 7 days，RT-PCR was performed to detect the mRNA expression level of BMP9 and VEGF，and real-time PCR was carried out to detect the mRNA ex－pression level of osteogenesis index osteocalcin（OC），osteoprotegerin（OPG），osteopontin（OPN）. Western blot was used to detect the protein expressions level of OPN. Alkaline phosphatase（ALP） activity was measured by quantitative and staining assay，and calcium deposition was determined by Alizarin Red S staining in each group. Results：BMP9 and VEGF co-expressed effectively in Ad-VEGF+Ad-BMP9 group. The mRNA level of OC，OPG，OPN and the protein level of OPN in Ad-BMP9 group and Ad-VEGF+Ad-BMP9 group were statistically higher than those of other groups（P<0.05），and were the highest in Ad-VEGF+Ad-BMP9 group（P<0.05）. The activity and staining assay of ALP，and the deposition of calcium in Ad-VEGF+Ad-BMP9 group were statistically higher than those in the other groups（P<0.05）. Conclusion：Combination of BMP9 and VEGF may promote osteogenic differentiation of C3H10T1/2 cells in vitro.

关键词

骨形态发生蛋白9血管内皮细胞生长因子成骨分化

Keywords

bone morphogenetic protein 9vascular endothelial growth factorosteogenesis

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