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目的制备主动靶向乳腺癌的pH响应负载多烯紫杉醇(docetaxel,DTX)的纳米粒子,并考察其理化性质、载药和药物缓释特征及对人乳腺癌MCF-7细胞的靶向和杀伤效果。方法采用纳米沉淀法和基于聚多巴胺(polydopamine,PDA)的表面修饰方法制备主动靶向MCF-7细胞的载DTX的叶酸(folic acid,FA)和PDA修饰的胆酸-聚乳酸-羟基乙酸共聚物纳米粒子(DTX-loaded CA-PLGA@PDA-PEG-FA/NPs);采用透射电镜观察纳米粒子的形貌,纳米粒度仪分析纳米粒子的粒径和zeta电位,X射线光电子能谱仪(XPS)分析纳米粒子表面修饰情况;采用透析法和高效液相色谱法研究纳米粒子的载药率、包封率以及体外释放曲线;采用激光扫描共聚焦显微镜和流式细胞仪分析负载荧光探针的纳米粒子的体外细胞摄取;采用MTT法研究载药纳米粒子对MCF-7细胞的存活率的影响。结果本研究制备的DTX-loaded CA-PLGA@PDA-PEG-FA/NPs呈“核-壳”结构,水合粒径为(166.4±3.9)nm,zeta电位为(-11.7±3.8)mV,载药量为(9.67±0.45)%,包封率为(88.32±3.10)%,在pH 5.0的释放介质中药物释放较在pH7.4的释放介质中快,XPS分析结果显示PDA和叶酸在纳米粒子表面的修饰,MCF-7细胞摄取的主动靶向的纳米粒子多于未连接主动靶向配体的纳米粒子,载药主动靶向纳米粒子的细胞毒性明显优于DTX的临床制剂泰素帝○R。结论主动靶向乳腺癌的pH响应的载药纳米粒子表现出良好的主动靶向性和MCF-7细胞杀伤效果。
OBJECTIVE: To prepare pH-responsive nanoparticles loaded with docetaxel (DTX), which targets active breast cancer, and investigate its physicochemical properties, drug-loaded and drug-release characteristics and its targeting and killing effect on human breast cancer MCF-7 cells effect. METHODS: DTX-loaded folic acid (FA) and PDA-modified cholic acid-poly (DL-lactic-co-glycolic acid) were prepared by nano-precipitation method and surface modification method based on polydopamine The morphology of the nanoparticles was observed by transmission electron microscopy. The particle size and zeta potential of the nanoparticles were analyzed by the nano-particle size analyzer. The X-ray photoelectron spectroscopy XPS) was used to analyze the surface modification of nanoparticles. The drug loading rate, entrapment efficiency and in vitro release curve of nanoparticles were investigated by dialysis method and high performance liquid chromatography (HPLC). The fluorescence intensity of loaded nanoparticles was analyzed by laser scanning confocal microscopy and flow cytometry Of the nanoparticles in vitro cell uptake; using MTT assay drug-loaded nanoparticles on the survival rate of MCF-7 cells. Results The DTX-loaded CA-PLGA @ PDA-PEG-FA / NPs exhibited a “core-shell” structure with a hydrated diameter of (166.4 ± 3.9) nm and a zeta potential of (-11.7 ± 3.8) mV , Drug loading (9.67 ± 0.45)% and entrapment efficiency (88.32 ± 3.10)%. The drug release in pH 5.0 release media was faster than that in pH 7.4 release media. The results of XPS analysis showed that PDA and folic acid The modification of the surface of nanoparticles, MCF-7 cells uptake more active nanoparticles than active ligand-free nanoparticles, the active drug targeting nanoparticles nanoparticles cytotoxicity was significantly better than DTX clinical preparations of Thai Prime Emperor ○ R. Conclusion The drug-loaded nanoparticles that actively target the pH response of breast cancer show good active targeting and MCF-7 cell killing effect.