2种粒径PLGA纳米粒细胞摄取动力学的比较

摘要

目的:比较包载有荧光香豆素-6(coumarin-6,C6)的2种粒径聚乳酸-羟基乙酸共聚物(plylactic acid-glycolic acid copolymer,PLGA)纳米粒,经肿瘤细胞的摄取动力学的差异.方法:以生物可降解材料PLGA为载体,采用乳化溶剂挥发法制备2种不同粒径的载有C6的PLGA纳米粒,以黑色素肿瘤细胞(B16)和脑胶质瘤细胞(U87)为细胞模型,分别与2种粒径的纳米粒孵育0.5,1,4,6和17 h,用荧光光度计检测细胞裂解液中C6的荧光值,计算得出2种细胞摄入的载药PLGA纳米粒的量(以C6的量表示),绘制入胞动力学曲线.应用GraphPad Prism程序处理2种细胞得出的数据,并应用T检验法分析各组数据之间的差异.结果:孵育0.5h后,2种粒径纳米粒在2种细胞中的摄取量均表现出显著差异;且随着时间延长,该差异加大.细胞摄取时间在6 ～17h时,粒径约为150 nm的PLGA纳米粒的入胞速率明显变缓.结论:在相同的给药浓度下,小粒径纳米粒入胞速率要快于较大粒径的纳米粒.大粒径的纳米粒比小粒径纳米粒的入胞量到达平衡快,且总的入胞量要少.%Objective:To compare the difference of intracellular kinetics between PLGA nanoparticles with zeta size of around 100 nm and 150 nm respectively.Methods:PLGA nanoparticles loaded with coumarin-6 was prepared by emulsion solvent evaporation method.Melanoma tumor cells (B16) and glioma cells (U87) were used as cellular uptake models.After incubating with nanoparticles for 0.5,1,4,6 and 17 h,the endocytosed nanoparticles was quantified by detecting the content of coumarin 6 in the cellular lysate by fluorospectrophotometer.The data obtained was analyzed by GraphPad Prism program and the difference was evaluated by using student's t-test.Results:After incubating for 0.5 h,the intracellular quantity of PLGA nanoparticles of 100 nm exhibited significant difference from the particles of 150 nm,and the difference was enlarged as time going.The intracellular rate of nanoparticles of 150 nm decreased obviously by the end of 6 h and after.Conclusion:PLGA nanoparticles of 100 nm intend to be endocytosed more easily and quickly than particles of 150 nm in both U87 cells and B16 cells.And the overall intracellular amount of PLGA nanoparticles of 150 nm is less than that of nanoparticles of 100 nm.