[Vascular endothelial growth factor/polylactide-polyethyleneglycol-polylactic acid copolymer/basic fibroblast growth factor mixed microcapsules in promoting angiogenic differentiation of rat bone marrow mesenchymal stem cells in vitro].

Objective: To observe the effect of vascular endothelial growth factor/polylactide-polyethyleneglycol-polylactic acid copolymer/basic fibroblast growth factor (VEGF/PELA/bFGF) mixed microcapsules in promoting the angiogenic differentiation of rat bone marrow mesenchymal stem cells (BMSCs) in vitro. 方法: 经全骨髓贴壁法分离培养 BMSCs 并传代。取第 3 代 BMSCs 行 Wright-Giemsa 染色及流式细胞术鉴定。利用复乳溶剂挥发法制备 VEGF/PELA/bFGF（A 组）、PELA/bFGF（B 组）、VEGF/PELA（C 组）及 PELA（D 组）微囊。对 PELA 微囊行降解性能及细胞毒性检测，VEGF/PELA/bFGF 混合微囊行 VEGF 及 bFGF 体外释放检测。取 4 组微囊缓释上清液分别与第 3 代 BMSCs 培养。培养 1、3、7、14、20 d 于倒置显微镜下观察细胞形态变化；21 d 时，行摄取 Dil 标记的乙酰化低密度脂蛋白（DiI-labeled acetylated low density lipoprotein，Dil-ac-LDL）及 FITC 标记的荆豆凝集素（FITC-labeled ulex europaeus agglutinin I，FITC-UEA-I）实验，观察细胞摄取能力；同时行 Matrigel 基质胶小管形成实验，定量分析小管形成指标（节点数、交叉点数、主干数及主干长度）。. Results: The isolated and cultured cells were identified as BMSCs. The degradation time of PELA was more than 20 days. There was no significant effect on cell viability under co-culture conditions. At 20 days, the cumulative release of VEGF in the mixed microcapsules exceeded 95%, and the quantity of bFGF exceeded 80%. The morphology of cells in groups A, B, and C were changed. The cells in groups A and B showed the typical change of cobble-stone morphology. The numbers of double fluorescent labeled cells observed by fluorescence microscope were the most in group A, and decreases from group B and group C, with the lowest in group D. The cells in groups A and B formed a grid-like structure on Matrigel. Quantitative analysis showed that the differences in the number of nodes, master junctions, master segments, and tot.master segments length between groups A, B and groups C, D were significant ( P 0.05). Conclusion: VEGF/PELA/bFGF mixed microcapsules have significantly ability to promote the angiogenic differentiation of rat BMSCs in vitro.