PEG化聚乳酸羟基乙酸纳米粒的抗黏性及HeLa细胞摄取

Anti-adhesive activity and uptake by HeLa cells of mPEG-PLGA nanoparticles

目的制备单甲氧基聚乙二醇-聚乳酸-羟基乙酸纳米粒(monomethoxy polyethylene glycol-poly-actic coglycolic acid-nanoparticles,mPEG-PLGA-NPs)并研究其理化性质及体外抗黏性能,考察人宫颈癌细胞HeLa对mPEG-PLGA-NPs的摄取能力。方法采用溶剂扩散法制备mPEG-PLGA-NPs,测定其平均粒径和zeta电位。采用黏蛋白结合法,考察mPEG-PLGA-NPs的体外抗黏性能。以香豆素-6(coumarin 6)为荧光标记物,通过共聚焦显微镜及HPLC法进行HeLa细胞的体外摄取研究。结果 mPEG-PLGA-NPs的平均粒径为(106.2±4.3)nm,Zeta电位为-(12.40±0.11)mV。PLGA-NPs的抗黏蛋白黏附率为35.5%,而mPEG-PLGA-NPs的抗黏蛋白黏附率为92.5%,比PLGA-NPs高3倍左右。相同孵育时间内,HeLa细胞对mPEG-PLGA-NPs的摄取量是PLGA-NPs的近2倍,HeLa细胞对PLGA-NPs和mPEG-PLGA-NPs的摄取量与孵育时间成明显的依赖关系。结论 mPEG-PLGA-NPs具有较强的抗黏性;HeLa细胞对mPEG-PLGA-NPs的摄取明显高于PLGA-NPs(P<0.01),表现出更好的细胞亲和性。

Objective To prepare mPEG-PLGA nanoparticles and discuss its properties and in vitro anti-adhesive activity and investigate the cellular uptake of them in human cervical cancer cells （HeLa）. Methods mPEG-PLGA-NPs were prepared by a solvent diffusion method. The particle size distribution and zeta potential of nanoparticles were measured by light scattering. The anti-adhesive activity of nanoparticles was evaluated by pig gastric mucin （PM） binding experiments. Coumarin 6 was incorporated into nanoparticles as fluorescent marker. The cellular uptake of PLGA-NPs and PEG-PLGA-NPs by HeLa cell lines was determined by laser scanning confocal microscope and HPLC. Results The mean size of mPEG-PLGA-NPs was （106.2±4.3） nm, Zeta potential was –（12.40±0.11） mV. The PM anti-adhesive rate of PLGA-NPs and PEG-PLGA-NPs was 35.5% and 92.5% respectively. The anti-adhesive activity of mPEG-PLGA-NPs was three times higher than PLGA-NPs. All nanoparticles showed low cytotoxicity. At the same incubation time, the uptake of mPEG-PLGA-NPs was two times higher than that of unmodified PLGA-NPs. The cellular uptake of PLGA-NPs and mPEG-PLGA-NPs was dependent on the incubation time. Conclusions mPEG-PLGA-NPs has much higher anti-adhesive activity. The uptake of mPEG-PLGA-NPs by HeLa cells is much higher compared to the PLGA-NPs（P〈0.01）. The mPEG-PLGA-NPs shows better cell affinity than PLGA-NPs.