包载姜黄素纳米胶束的制备与体外抗肿瘤评价

Preparation and in vitro antitumor evaluation of curcumin-loaded micelles

目的:构建载姜黄素纳米胶束,体外评价其对C6脑胶质瘤细胞的生长抑制作用。方法:合成新型十一烯酸-接枝-ε-多聚赖氨酸(ε-PLL-UNA)聚合物,并对其结构进行表征,采用芘荧光探针法对其临界胶束浓度进行测定;以ε-PLL-UNA为材料,采用透析法制备载姜黄素纳米胶束,以动态光散射和透射电镜对其粒径、形态进行表征,以动态透析法测定药物释放行为,以激光共聚焦显微镜观察体外细胞摄取性,以肿瘤球模型考察其体外抗肿瘤作用。结果:~1H-NMR和FT-IR结果表明ε-PLL-UNA聚合物成功合成,其临界胶束浓度为0.19g/L,能自发组装成胶束;载姜黄素纳米胶束载药量能达到12.22%±2.13%、包封率则高达85.12%±3.64%,平均粒径为(60.6±2.1)nm、Zeta电位为(28.2±5.6)m V,具有球形微观结构;该纳米胶束48h释放84%的姜黄素,体外快速被C6细胞摄取;与姜黄素溶液相比,该纳米胶束能明显抑制胶质瘤细胞球的生长。结论:ε-PLL-UNA聚合物胶束对姜黄素具有较高的载药量,粒径小于100nm、分布均匀,体外缓慢释放药物,提高了C6细胞对姜黄素的摄取,而且对C6细胞球具有有效的杀伤作用。

Objective:To prepare a novel polymer micelles encapsulating curcumin(CUR)and evaluate itsanti-tumor effect on glioma in vitro.Methods:A novel polymer,undecenoic acid-grafted-ε-polylysine(ε-PLLUNA),was synthesized,and its chemical structure was confirmed by1H-NMR and FT-IR.The CAC value ofε-PLL-UNA polymer was also detected by pyrene fluorescence probe.CUR-loaded micelle was prepared bydialysis method usingε-PLL-UNA as materials,and its particle size and morphology were also studied underdynamic light scattering(DLS)and transmission electron microscope(TEM),respectively.Furthermore,in vitrodrug release profiles from CUR-Micelles were explored by dynamic dialysis method.Finally,the cellular toxicityagainst C6cells spheroids and the cellular uptake of CUR-Micelles were evaluated.Results:ε-PLL-UNA polymerwas successfully synthesized and able to self-assemble into micelles above its CAC value of0.19g/L.CURloadedmicelle had a mean diameter of(60.6±2.1)nm,and zeta potential of(28.2±5.6)mV,exhibiting the sphericalshape determined by TEM.Drug loading content and drug loading efficiency for CUR-loaded micelle were highup to12.22%±2.13%and85.12%±3.64%,respectively.About84%of CUR were released from the micelles in48hours.CUR-loaded micelle can promote the cellular uptake of its encapsulated CUR by C6cells,displaying asignificantly higher cytotoxicity against C6cells.Besides,the growth of C6cells spheroids was significantly inhibitedby CUR-loaded micelle.Conclusion:CUR is efficiently encapsulated inε-PLL-UNA micelles with a particlesize of less than100nm,which improved the cellular uptake of C6cells.The sustained-release of CUR fromCUR-loaded micelle is also observed.More importantly,CUR-Micelles has superior growth inhibition against C6cells spheroids.