高分子修饰细菌纤维素细胞相容性的初步研究

Primary research on biocompatibility of poly lactic acid modified bacterial cellulose

目的筛选适于平滑肌细胞附着生长的聚乳酸-共-聚乙醇酸(PLGA)修饰的细菌纤维素材料。方法不同比例PLGA修饰的细菌纤维素材料分为5组:单纯PLGA材料(50P组);经PLGA修饰的细菌纤维素材料组则包括聚乳酸(PLA)∶聚乙醇酸(PGA)=50∶50的50PB组、PLA∶PGA=75∶25的75PB组和PLA∶PGA=90∶10的90PB组;单纯细菌纤维素为空白对照组。将平滑肌细胞接种于不同组别的材料上培养7 d,扫描电镜观察细胞的生长状态,并用荧光染色计数活/死细胞,CCK8法测定材料上的细胞增殖情况并绘制增殖曲线。不同材料上细胞数量及增殖数据用单因素方差分析比较、两两比较用SNK检验。结果接种后第3天的扫描电镜可见细胞均能在各组材料上成功附着并生长,但75PB组和空白对照组细胞生长形态较差。活/死细胞染色结果显示,平滑肌细胞可以在各组材料中均匀的分布以及生长,活细胞计数各组差异无统计学意义(F=1.454,P>0.05)。细胞生长曲线检测显示,接种后3 d及5 d,平滑肌细胞在各组材料上的增殖量差异无统计学意义(3 d F=1.672,P>0.05;5 d F=1.19,P>0.05);接种后7 d,50PB组和空白对照组的平滑肌细胞增殖优于90PB组及75PB组(F=13.328,P<0.01)。结论PLA/PGA比例为50∶50的PLGA修饰细菌纤维素后的材料细胞相容性更优,可以成为用于组织修复的生物材料。

Objective To investigate the biocompatibility of poly lactic-co-glycolic acid(PLGA)modified bacterial cellulose using smooth muscle cells.Methods Different proportions of PLGA modified bacterial cellulose were divided into five groups:the pure PLGA group(the 50P group),bacterial cellulose modified by PLA∶PGA=50∶50 as the 50PB group,bacterial cellulose with poly-lactic acid(PLA)∶poly-glycolic acid(PGA)=75∶25 as the 75PB group and bacterial cellulose with PLA∶PGA=90∶10 as the 90PB group;the pure bacterial cellulose as the control group.The smooth muscle cells were seeded to different kinds of materials and cultured for 7 d.The situation of smooth muscle cells on different materials was observed by electronic migroscope(SEM)and determined by live/dead staining.The cell proliferation on the materials was determined by cell counting kit-8(CCK-8)test.The data of cell survival rate and proliferation rate were analyzed by one-way ANOVA and SNK test.Results The smooth muscle cells in all the groups adhered well to the materials on the 3 rd day according to SEM observation.The live/dead staining result showed that smooth muscle cells of all the groups distributed uniformly and grow flourishly(F=1.454,P>0.05).The CCK-8 data indicated that the growth of smooth muscle cells in different groups had no difference on the 3 rd day and 5 th day(3 d F=1.672,P>0.05;5 d F=1.19,P>0.05).On the 7 th day the cells both in the 50PB group and the control group showed superior proliferation to those in the 90PB group and the 75PB group(F=13.328,P<0.01).Conclusion The biocompatibility of PLGA(PLA∶PGA=50∶50)modified bacterial cellulose is better than other PLGA proportion modified bacterial cellulose,which is potential biomaterial for tissue repair.