载吲哚菁绿和多柔比星自组装胶束的构建及体外抗肿瘤及其转移的评价

Preparation and evaluation of anti-tumor metastasis of self-assembly micelles loaded with indocyanine green in vitro

本研究通过p H敏感的腙键共价连接疏水性化疗药物多柔比星(doxorubicin,DOX)和亲水性抗肿瘤转移的低分子量肝素(low molecular weight heparin,LH)合成了低分子量肝素-多柔比星胶束(LH-DOX),并物理包载光热剂吲哚菁绿(indocyanine green,ICG),以透析法制备了具有p H敏感的载吲哚菁绿的低分子量肝素-多柔比星聚合物胶束(LH-DOX/ICG)。该胶束粒径为(148.7±2.1)nm,在电镜下呈形态规则的球形,粒径均一,并具有良好的稳定性和生物相容性。体外释放结果显示,DOX的释放具有p H敏感性,ICG有一定的缓释效果。ICG包载于胶束后与游离ICG具有相似的光热效应。一方面,划痕愈合实验显示,在激光照射下LH-DOX/ICG胶束具有良好的抗肿瘤转移能力;另一方面,细胞凋亡及细胞毒性实验表明,在激光照射下该胶束诱导黑色素瘤B16F10细胞凋亡的能力和对细胞毒性作用明显增加。因此,LH-DOX/ICG胶束具有良好的前景,用于光疗-化疗联合治疗黑色素瘤。

In this study,we used low molecular weight heparin（LH） as hydrophilic sides,doxorubicin（DOX） as hydrophobic sides,and indocyanine green（ICG） as a photosensitive drug to prepare p H-sensitive self-assembled polymeric micelles（LH-DOX/ICG）.The micelles were prepared by dialysis,and evaluated for stability,p H sensibility in vitro,hemolytic test and photo-thermal effect.Wound healing test was used to evaluate the anti-metastatic effects.MTT assay and apoptosis detection kit were used to evaluate the cytotoxicity of micelles against melanoma B16 F10 cells.The size of the micelles was（148.7 ± 2.1） nm and the zeta potential was（-30.7 ± 1.1） m V.The drug-loading content of DOX and ICG were 11.82% and 5.59%,respectively.The micelles exhibited spherical in shape,fairly uniform size.The micelles were stable within 48 h in 50% fetal bovine serum phosphate buffer.The release of DOX was p H-sensitive,while the release of ICG was sustained.The micelles were biocompatible and safe as indicated by the hemolytic test.In vitro photo-thermal effect indicated LH-DOX/ICG micelles had similar photo-thermal effect to the free ICG.Wound healing test showed that the micelles had a good ability to inhibit tumor migration with laser irradiation.MTT assay and cell apoptosis assay indicated that LH-DOX/ICG micelles displayed more efficient anti-tumor effect after near infrared laser compared to LH-DOX micelles.LH-DOX/ICG micelles are promising for the therapeutic effect of phototherapy and chemotherapy in combination for melanoma.