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合成聚乙二醇-聚己内酯-聚乙烯亚胺(PEG-PCL-PEI)三嵌段聚合物载体材料,制备包载大黄酸(RH)的PEGPCL-PEI纳米粒(PPP-RH-NPS),并评价其理化性质和体外生物学特性。通过开环聚合和麦克尔加成反应合成得到PEG-PCL-PEI聚合物,相对分子质量为9.5×10~3,临界胶束浓度为0.723 nmol·L~(-1)。包载RH制备得到PPP-RH-NPS,外观浅黄、乳光明显,透射电镜观察纳米粒分散均匀圆整无团聚,粒径为(118.3±3.6)nm,PDI为(0.19±0.08),Zeta电位为(6.3±1.5)mV,包封率为(93.64±5.28)%,载药量为(8.57±0.53)%。透析法考察PPP-RH-NPS体外释药特征,48 h内累积释放率为75.92%,释药曲线符合Higuchi模型方程:Q=0.121 6t~(1/2)+0.069 5(R~2=0.887 4),呈缓释特性。选用兔红细胞考察其溶血率,MTT法评价其对HK-2细胞的生物安全性,在0~0.05 mmol·L~(-1)不会引起红细胞溶血和细胞毒性。流式细胞仪考察其摄取效率,PPP-RH-NPS可被细胞迅速内吞,摄取效率高,30 min内摄取完全,经激光共聚焦显微镜观察,PPP-RH-NPS能够从溶酶体进入细胞质,具有溶酶体逃逸特性。因此,该研究成功合成PEG-PCLPEI聚合物和制备得到PPP-RH-NPS,粒径分布均匀,包封率及载药量较高,摄取迅速,具有缓释特征、溶酶体逃逸特性、优良的生物安全性,是一种良好研究前景的新型纳米制剂。
Polyethylene glycol-polycaprolactone-polyethylenimine (PEG-PCL-PEI) triblock polymeric carrier material was synthesized to prepare rhein (PEG) RH-containing PEGPCL- ), And to evaluate its physical and chemical properties and in vitro biological characteristics. The PEG-PCL-PEI was synthesized by ring-opening polymerization and Michael addition reaction. The relative molecular mass was 9.5 × 10 ~ 3 and the critical micelle concentration was 0.723 nmol·L -1. The PEG-RH-NPS was prepared by the inclusion of RH. The appearance of the sample was light yellow and the opalescence was obvious. The nanoparticles were uniformly distributed without rounding and the size was (118.3 ± 3.6) nm. The PDI was (0.19 ± 0.08) (6.3 ± 1.5) mV, the entrapment efficiency was (93.64 ± 5.28)% and the drug loading was (8.57 ± 0.53)%. The in vitro release characteristics of PPP-RH-NPS were investigated by dialysis method. The cumulative release rate was 75.92% within 48 h. The release curve was in accordance with Higuchi model equation: Q = 0.121 6t 1/2 (1/2) +0.069 5 (R 2 = 0.887 4), was slow release characteristics. Rabbit erythrocytes were used to investigate the hemolysis rate. The biologic safety of HK-2 cells was evaluated by MTT assay. Hemolysis and cytotoxicity of erythrocytes were not induced at 0 ~ 0.05 mmol·L -1. The uptake efficiency of PPP-RH-NPS was examined by flow cytometry. PPP-RH-NPS was quickly endocytosed by the cells and the uptake efficiency was high, which was completely absorbed within 30 min. PPP-RH-NPS was able to enter the cytoplasm from lysosomes by laser confocal microscopy , With lysosomal escape properties. Therefore, this study successfully synthesized PEG-PCLPEI polymer and preparation of PPP-RH-NPS, particle size distribution, encapsulation efficiency and drug loading is high, rapid intake, with sustained release characteristics, lysosomal escape characteristics, excellent Of biological safety, is a good research prospects of new nano-formulations.