pH敏感电荷反转型姜黄素纳米粒子的制备及体外评价

Preparation and in vitro evaluation of pH-sensitive charge-reversal nanoparticles loaded with curcumin

目的制备pH敏感电荷反转型姜黄素纳米粒子(PCE/Cur NPs),优化制备工艺并考察PCE/Cur NPs的理化性质及其对黑色素瘤(B16)细胞的抑制作用。方法在正电性材料甲氧基聚乙二醇-聚己内酯-聚乙烯亚胺(MPEG-PCL-PEI,PCE)的基础上键合1,2-环己二酸酐使其变成负电性材料,制备带负电的PCE/CurNPs,该纳米粒子具有pH值依赖性,pH值>7时,PCE/Cur NPs不发生化学变化,显示负电性,pH值<6时,PCE/Cur NPs发生化学反应,水解掉1,2-环己二酸,显示正电性。考察了其形态、粒径、载药量、包封率与体外释放等理化特性;通过MTT法、划痕实验进行检测,验证PCE/Cur NPs对B16细胞的抑制作用、抗增殖侵袭能力。结果获得了PCE/Cur NPs,透射电子显微镜下观察所得纳米粒子形态外观圆整,大小均匀,无黏连,平均粒径为(80±5)nm;包封率为(90.0±2.0)%;载药量为(8.0±1.0)%;48 h时,PCE/Cur NPs释放趋于恒速,释放缓慢,累积释放达到(69.2±5.2)%(pH 7.4)与(71.2±4.3)%(pH 5.0);细胞实验显示,PCE/Cur NPs能够显著抑制B16细胞的生存和增殖能力,具有剂量和时间依赖性。结论成功制备了PCE/CurNPs,对抑制B16细胞增殖效果良好,为开发智能型抗肿瘤给药系统提供了理论依据。

Objective To prepare charge-reversal pH-sensitive nanoparticles loaded with curcumin(PCE/Cur NPs), and investigate the optimizing technology, physicochemical characterizations, and inhibitory effect on B16 cell. Methods The β-carboxyl amidized cationic MPEG-PCL-PEI polymers(PCE) were negatively charged, which were prepared into the negative PCE/Cur NPs with pH dependence. When pH > 7, there was no charge-reversal. When pH < 6, the β-carboxyl amides were hydrolyzed rapidly into corresponding amines. As a result, PCE/Cur NPs became positively charged again. The obtained PCE/Cur NPs were characterized by detection of particle size, morphology study, drug loading, encapsulation efficiency, and release study. The effect of anti-migratory and anti-invasive actions of PCE/Cur NPs on B16 cell was investigated using MTT assays and wound healing test. Results PCE/Cur NPs dependent on pH charge inversion were successfully prepared. The obtained PCE/Cur NPs were round, and the size was uniform, the adhesion was not found. The results showed that the prepared PCE/Cur NPs had the highest DL(8.0 ± 1.0)%, EE(90.0 ± 2.0)%, mean particle size of(80 ± 5) nm, and zeta potential of(-35 ± 5) m V. Within 48 h, the accumulative release rate was(69.2 ± 5.2)%(pH 7.4) and(71.2 ± 4.3)%(pH 5), respectively, and then PCE/Cur NPs released slowly. These results by MTT assay and wound healing assay indicated that PCE/Cur NPs not only inhibited the proliferation of B16 cells in a concentration-and time-dependent manner, but also can induce apoptosis. Conclusion PCE/Cur NPs were prepared successfully, which might have great potential application in drug delivery system.