PLGA 、壳聚糖纳米粒的制备及细胞毒性研究

摘要

AIM:To prepare PLGA and chitosan ( CS) nanoparticles and to evaluate their cytotoxicity to periodontal ligament cells ( PDLCs) .METHODS:PLGA and CS nanoparticles were prepared by emulsion solvent e-vaporation method cross-linking of CS with sodium tripolyphosphate respectively.The nanoparticles were characterized by Zetasizer 3000HS and transmission electron microscope.Passage 3 -5 PDLCs were cultured with PLGA and CS nanoparticles at 0(the control), 0.5 mg/mL, 1 mg/mL and 2 mg/mL respectively.The PDLCs were subjected to MTT assay at 24, 48, 72 h of culture.RESULTS:PLGA nanoparticles were uniform and spherical, and the mean particle size and dispersive coefficient were 188.10 ±2.86 nm and 0.55 ±0.04, respectively.CS nanoparticles were relatively uniform and aspheric, the mean particle size and dispersive coefficient were 250.97 ±30.86 nm and 0.50 ±0.07, re-spectively.The OD values of 2 mg/mL PLGA group and 1 mg/mL CS group were lower than that of the control (P<0.05) at 48 h of culture.At other times points, no significant difference was found among different groups (p>0.05).The relative growth rate of PDLCs in all groups was all higher than 85%.The toxicity grading of all groups was 0 or 1.CONCLUSION:PLGA and CS nanoparticles are nontoxic to PDLCs, according to ISO10993-5.%目的：制备大小均一的PLGA、壳聚糖纳米粒并研究其对牙周膜细胞的毒性。方法：采用乳化溶剂挥发法制备PLGA纳米粒；离子交联法制备壳聚糖纳米粒；用Zetasizer 3000 HS检测纳米粒的粒径和分散系数，透射电镜观察纳米粒的形态。取3～5代人牙周膜细胞，分别与0（对照组）、0．5、1、2 mg／mL的PLGA和壳聚糖纳米粒以及1 mg／mL普通粒径的PLGA和壳聚糖粉末共同培养；分别于培养24、48、72 h后用MTT法检测各组细胞的相对增殖率，评价PLGA、壳聚糖纳米粒的细胞毒性。结果：透射电镜观察：PLGA纳米粒呈球形，壳聚糖纳米粒类似球形，两者大小较均匀、边缘清晰。 Zetasizer 3000HS检测显示：PLGA纳米粒的粒径为（188．10±2．86） nm，分散系数为0．55±0．04；壳聚糖纳米粒的粒径为（250．97±30．86） nm，分散系数为0．50±0．07。毒性实验结果：培养48 h时，PLGA、壳聚糖粉末组以及2 mg／mL PLGA、1 mg／mL壳聚糖纳米粒组的细胞增殖率均低于空白对照组（P＜0．05），其他各时间点各组间均无统计学差异（ P＞0．05）；各组各时间点的细胞相对增殖率均大于85％，细胞毒性为0或1级。结论：根据ISO10993－5，PLGA和壳聚糖纳米粒对牙周膜细胞无毒性。