载阿霉素的大黄酸偶联物胶束的制备及其细胞毒性评价

Preparation and Cytotoxicity Evaluation of Doxorubicin-loaded Rhein Conjugate Micelles

目的制备载阿霉素(doxorubicin, DOX)的TPGS修饰的羧甲基壳聚糖-大黄酸(TPGS-CR)偶联物胶束(DOX/TPGS-CR胶束),考察其体外对人乳腺癌MCF-7细胞的毒性。方法以TPGS-CR偶联物为载体材料,利用透析法制备DOX/TPGS-CR胶束,使用动态激光粒径测定仪测定其粒径,紫外分光光度法测定载药量和包封率;在pH 7.4的PBS中进行了体外释放研究,计算了DOX的累计释放率,绘制了累计释放曲线;MTT法检测其对人乳腺癌MCF-7细胞毒作用,计算细胞的存活率。结果 DOX/TPGS-CR胶束的平均粒径为(151.3±1.8)nm,PDI为0.129±0.020,电位为(–26.9±0.8)mV,载药量为(23.16±0.01)%,包封率为(55.00±0.04)%。DOX/TPGS-CR胶束在pH 7.4的PBS中,24 h时累计释放DOX仅有(35.73±2.21)%,而游离DOX·HCl在6h时累计释放达到了(90.25±6.20)%;DOX/TPGS-CR胶束浓度在5~20μg·mL–1内,尤其在48h时,对MCF-7细胞杀伤能力强于DOX·HCl。结论 DOX/TPGS-CR胶束载药量良好,粒径小,分布均匀,具有缓释特征;载体材料TPGS-CR偶联物毒性小,安全性高,DOX/TPGS-CR胶束对MCF-7细胞有较强的杀伤作用。

OBJECTIVE To prepare doxorubicin(DOX)-loaded TPGS modified carboxymethyl chitosan-rhein(TPGS-CR) polymeric micelles(DOX/TPGS-CR PMs), and study its cytotoxicity in human breast cancer MCF-7 cells in vitro. METHODS DOX/TPGS-CR PMs were prepared by dialysis method using TPGS-CR conjugate. The particle size of DOX/TPGS-CR PMs was detected by dynamic light scattering(DLS), the entrapment efficiency(EE) and the drug loading-capacity(DL) were studied by Ultraviolet spectrophotometry. The vitro release studies were performed in PBS(pH 7.4), the cumulative release rate of DOX was calculated, and the cumulative release profile was plotted. The cytotoxicity of DOX/TPGS-CR PMs in human breast cancer MCF-7 cells was detected by MTT assay, and the survival rate of the cells was calculated. RESULTS The average particle size and polydispersity coefficient of DOX/TPGS-CR PMs were(151.3±1.8)nm and 0.129±0.020, respectively. The zeta potential of DOX/TPGS-CR PMs was(–26.9±0.8)mV. The DL and EE of DOX/TPGS-CR PMs were(23.16±0.01)% and(55.00±0.04)%, respectively. Cumulative release of DOX in PBS(pH 7.4) from DOX/TPGS-CR PMs was only about(35.73±2.21)% at 24 h, while the cumulative release of free DOX·HCl reached(90.25±6.20)% at 6 h. The cytotoxicity of DOX/TPGS-CR PMs was better than doxorubicin hydrochloride(DOX·HCl) in the concentration range of 5-20 μg·mL–1, especially at 48 h. CONCLUSION The DL of DOX/TPGS-CR PMs is good, and the particle size is small and uniform. DOX/TPGS-CR PMs have a sustained-release characteristic. TPGS-CR conjuage is less toxic and safe. DOX/TPGS-CR PMs display cytotoxicity significantly in MCF-7 cells.