血管内皮生长因子对骨髓间充质干细胞向软骨细胞分化的影响

Effect of VEGF on chondrogenic differenciation of human bone marrow derived mesenchymal stem cells

[目的]探讨外源性血管内皮生长因子(VEGF)对人骨髓来源间充质干细胞(MSCs)增殖、向成软骨方向定向分化等生物学行为的影响及其机理,为MSCs构建组织工程化软骨奠定基础.[方法]取人骨髓来源间充质干细胞体外培养,将传代后的细胞分别置于含有800ng/ml和1600ng/mlVEGF的诱导培养基中进行培养,将具有促进细胞增殖分化及抑制多种炎性介质活性等多重生物学效应的TGF-β1以10ng/ml浓度配置无血清培养基诱导同组MSCs作阳性对照,观察细胞的增殖,分化。用Ⅱ型胶原免疫组化染色评价不同诱导因素下的软骨基质合成情况。[结果]所有细胞显示了良好的增殖能力,暴露于外源性血管内皮生长因子下的MSCs保持了良好的生长活性,并开始向软骨表型分化,与未添加诱导因子组细胞相比差异有显著性意义,不同浓度VEGF组的细胞合成软骨基质的能力无差异,但均不如阳性对照组。[结论]外源性VEGF具有诱导体外培养的人MSCs向软骨表型分化的能力,但与TGF-β1的诱导能力相比有差距。提示VEGF在MSCs来源的软骨修复过程中充当诱导信息提供者的角色。

[ Objective] To study the effect of vascular endothelial growth factor （VEGF） on proliferation of human bone marrow derived mesenchymal stem cells （h-MSCs） and its chondrogenic differenciation and to provide a basis for the tissue engineering technology of articular cartilage. [ Method] The hMSCs were cultured in vitro as usual and its passage was induced the by medium containing 800 ng/ml or 1600 ng/ml the VEGF MSCs the cultured in the medium containing 10 ng/ml the TGF β-1 which the had multiple the biological efficacies the to enhance the proliferation the of MSCs the and to the inhibit the activities of the inflammatory factors, acted as a positive control. The induced MSCs were used to observe its proliferation and differenciation. The synthesis of chondrocyte extracellular matrix in different induce-factor was measured by immunohisto-chemical staining. [Result] MSCs cultured wit different induced factors proliferated well, MSCs induced by VEGF medium showed a well growth activity and chondrogenic differenciation, but MSCs cultured in TGFβ1-containing medium induced more chondrocyte matrix than that in VEGF medium. [ Conclusion] VEGF induces chondrogenic differentiation of MSCs in vitro, but MSCs cultured in TGFβ1 medium induces more chondrocyte matrix It is suggested that VEGF acts as donor of inducer message cartilage repair by MSCs.