碱性成纤维细胞生长因子缓释微球对成骨细胞作用的实验研究

Effect of controlled release bFGF microspheres on osteoblasts

目的 旨在探讨碱性成纤维细胞生长因子 (bFGF)缓释微球生物活性保存情况及其对于成骨细胞的作用。 方法 将bFGF、bFGF -聚乳酸 -聚乙二醇共聚物 (PELA)微球和bFGF -聚乳酸 -羟基乙酸共聚物 (PLGA)微球加入成骨细胞培养液中 ,用细胞计数法、四甲基偶氮唑盐微量反应比色法 (MTT法 )、流式细胞仪观察细胞增殖情况 ,并检测成骨细胞上清液中骨钙素 (BGP)含量。 结果 培养 1d后 ,四组细胞计数、吸光度 (A)值差异均无显著性意义 ;4 ,6d时PLGA微球组细胞计数和A值明显优于其余三组 ;培养 6 ,8d后 ,bFGF组值仍然高于PELA微球组 ,但差异无显著性意义。流式细胞仪检测显示 ,培养 2d后 ,bFGF组的G2 M +S期百分数最高 ,4 ,8d后PLGA微球组G2 M +S期百分数最高。成骨细胞上清液中BGP含量 ,PLGA微球组最高 ,其次为PELA微球组。 结论 bFGF -PELA微球效果不佳 ,制备工艺需要改进 ;bFGF -PLGA微球通过较长时间持续释放活性bFGF ,明显促进成骨细胞增殖和分化。

Objective To investigate the bioactivities of controlled release bFGF microspheres (Ms) and their effects on the cultured osteoblasts. Methods The secondary cultured osteoblasts were divided into four groups according to the different ingredients being added to the DMEM culture medium, ie, control group,bFGF group, bFGF-PLGA-Ms group and bFGF-PELA-Ms group. The proliferation of the cultured osteoblasts was measured with cell counting method, MTT method and flow cytometry. The content of bone BGP secreted by osteoblast was also measured with RIA method. Results The in vitro cellular study showed no significant difference in the cell number and cell viability of four groups one day after plate culture.The cell number and cell viability in the bFGF-PLGA -Ms group were more than those in other three groups four and six days after plate culture. The cell number and cell viabilitythose in the bFGF group were more than those in the bFGF-PELA-Ms group six and eight days after plate culture with insignificant difference. The flow cytometrical examination showed that the G 2/M+S percentage in the bFGF group reached the highest two days after plate culture and the G 2/M+S percentage in the bFGF-PLGA-Ms group went the highest four and eight days after plate culture. Among four groups, the content of BGP in the bFGF-PLGA-Ms group was the highest and the bFGF-PELA-Ms group the next. Conclusions The effect of bFGF-PELA-Ms is not satisfactory,as indicates that the manufacturing method needs improving. However,the bFGF-PLGA-Ms can promote the proliferation and differentiation of the osteoblasts through a long period of controlled release of bFGF.