肺癌细胞膜涂层的DOX和IR820的光热敏感型仿生脂质体用于癌症的化学-光热-光动力治疗

Photothermal-sensitive biomimetic liposomes coated with DOX and IR820 for chemo-photothermal-photodynamic therapy of cancer in lung cancer cells

本研究构建以多柔比星(doxorubicin,DOX)为模型药物、新吲哚菁绿(IR820)为光敏剂、温度敏感脂质体(temperature sensitive liposomes,TSL)为载体,H460-NCI人大细胞肺癌细胞膜涂层的光热敏感脂质体(DOX-IR820-TSL@CCM),用于化学-光热治疗-光动力治疗的多途径高效肿瘤靶向作用。采用逆向蒸发法制备DOX-IR820-TSL,通过裂解、破碎、离心等方法制备癌细胞膜(cancer cell membrane,CCM),采用纳米膜挤压法制备DOX-IR820-TSL@CCM;并对上述脂质体进行表征。最终确定DOX-IR820-TSL的载药条件如下:有机相与水相比例为4.02,二棕榈酰磷脂酰胆碱(dipalmitoyl-sn-glycero-3-phosphocholine,DPPC)用量为10.04 mg,药脂比为0.12DOX-IR820-TSL显著相变温度(T_(m))为43.05℃,DOX-IR820-TSL@CCM平均粒径为153.4 nm,PDI为0.279,zeta电位为-26.2 mV。透射电子显微镜图像在图中显示出均匀的球形结构和半透明膜层。在近红外光照射下,药物释放率达到了63.98%,具有可调节的光热转化能力和产生活性氧的能力。通过SDS-PAGE电泳、Western blot、细胞毒性实验和细胞摄取实验,证明了细胞膜涂层的设计能够很好地保留CD47、N-cadherin、CD44、CD326等相关功能蛋白使DOX-IR820-TSL@CCM具备良好的免疫逃逸、同源黏附和同源靶向作用。本文制备了具有伪装特性和肿瘤靶向性的DOX-IR820-TSL@CCM,可作为一种协同治疗诊断平台用于未来的肺癌治疗。所有动物研究方案均已通过黑龙江中医药大学动物伦理委员会批准(编号为:2022121011)。

In this study,doxorubicin (DOX) was used as the model drug,new indocyanine green (IR820) as the photosensitizer,and temperature sensitive liposomes (TSL) as the carrier.H460-NCI photoheat-sensitive liposomes coated with cell membrane of human cell lung cancer (DOX-IR820-TSL@CCM) for highly effective multi-pathway tumor targeting in chemical-photothermal therapy and photodynamic therapy.DOX-IR820-TSL was prepared by reverse evaporation,cancer cell membrane (CCM) was prepared by lysis,crushing and centrifugation,and DOX-IR820-TSL@CCM was prepared by nanomembrane extrusion.The drug-loading conditions of DOX-IR820-TSL were finally determined:the ratio of organic phase to aqueous phase was 4.02,the dosage of dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) was 10.04 mg,and the lipid ratio was 0.12,and the significant phase transition temperature (T_m) of DOX-IR820-TSL was 43.05℃.The average particle size of DOX-IR820-TSL@CCM was 153.4 nm,the PDI was 0.279,and the zeta potential was-26.2 mV.The transmission electron microscope (TEM) image shows a homogeneous spherical structure and a translucent film layer.Under nearinfrared irradiation,the drug release rate reaches 63.98%,which has adjustable photothermal conversion capacity and the ability to generate reactive oxygen species.Through SDS-PAGE electrophoresis,Western blot,cytotoxicity experiments and cell uptake experiments,it was proved that the design of cell membrane coating can well retain CD47,N-cadherin,CD44,CD326 and other related functional proteins,so that DOX-IR820-TSL@CCM has good immune evasion,homologous adhesion and homologous targeting.In this paper,DOX-IR820-TSL@CCM with camouflage properties and tumor targeting properties were successfully prepared,which can be used as a promising synergistic therapeutic diagnostic platform for future lung cancer treatment.All animal research programs have been approved by the Animal Ethics Committee of Heilongjiang University of Chinese Medicine (number:2022121011).