碱性成纤维细胞生长因子缓释微球对雪旺细胞作用的实验研究

目的 探讨碱性成纤维细胞生长因子（bFGF）缓释微球对雪旺细胞的作用。方法 培养雪旺细胞，将bFGF、bFGF-聚乳酸-羟基乙酸共聚物微球（PLGA微球）和bFGF-聚乳酸-聚乙二醇共聚物微球（PELA微球）分别加入三个组的雪旺细胞培养液中。用细胞计数法测定雪旺细胞的数量，四甲基偶氮唑盐微量反应比色法（MTT法）测定雪旺细胞的活力，流式细胞仪测定雪旺细胞的细胞周期。结果 培养1、2d时，bFGF组的雪旺细胞计数、活力明显高于PLGA微球组和PELA微球组；培养3、4d时，bFGF组和PLGA微球组的雪旺细胞计数、活力明显高于PELA微球组；培养6、8d时．PLGA微球组的雪旺细胞计数、活力明显高于bFGF组和PELA微球组。流式细胞仪检测结果显示，培养2d后，bFGF组的Gz／M＋S期百分数最高；培养4、8d后，PLGA微球组的G2／M＋S期百分数最高，差异具有统计学意义。结论 游离bFGF对雪旺细胞促分裂增殖作用效应期短，而PLGA微球能在较长时期内促进雪旺细胞的分裂增殖，PELA微球虽然达到了缓释的性能要求，但释放出的bFGF的生物活性受到了破坏。

Effect of Controlled Release Microspheres Incorporating bFGF on Schwann Cells

Objective To investigate the effects of controlled release microspheres(Ms) incorporating bFGF on the cultured Schwann cells.Methods The secondary cultured Schwann cells were divided into three groups according to the different ingredients being added to the DMEM culture medium: bFGF group,and bFGF-PLGA-Ms group,and bFGF-PELA-Ms group.At different times after culture,the proliferative Schwann cells were collected from three groups individually.Then the number of Schwann cells was measured with cell counting method,the viability of Schwann cells was measured with MTT method and the cell cycle of Schwann cells was measured with flow cytometry.Results The in vitro cellular study showed that 1,2 days after plate culture,the number of cells and the cell viability of bFGF group were significantly larger than those of bFGF-PLGA-Ms group and bFGF-PELA-Ms group.3,4 days after plate culture,the number of cells and the cell viability of bFGF group and bFGF-PLGA-Ms group were significantly larger than those of bFGF-PELA-Ms group.6,8 days after plate culture,the number of cells and the cell viability of bFGF-PLGA-Ms group was significantly larger than those of bFGF group and bFGF-PELA-Ms group. For the flow cytometry examination,2 days after plate culture,the G_2/M S percentage of bFGF group was the highest,and 4,8 days after plate culture,the G_2/M S percentage of bFGF-PLGA-Ms group was the highest.Conclusion Free bFGF can promote the proliferation of Schwann cells in a short period,while bFGF-PLGA-Ms can promote the proliferation of Schwann cells in a long period because of the controlled release of bFGF from microspheres.bFGF-PELA-Ms meets the property requirement of controlled release,but the biological activity of released bFGF is destroyed partially.