多巴胺包裹的载药纳米金粒子在骨肉瘤光热治疗和化疗协同治疗中应用的实验研究

Experimental study on application of dopamine modified gold nanoparticles loaded with DOX for chemo-photothermal synergistic therapy in osteosarcoma

目的:探究负载化疗药物阿霉素(doxorubicin,DOX)的聚多巴胺包裹纳米金粒子用于化疗与光热协同治疗在骨肉瘤中的应用效果。方法:分别合成纳米金粒子(Au)、聚多巴胺包裹的纳米金粒子(Au@PDA)及载药后的纳米粒子(Au@PDA@DOX)。利用高倍透射电镜表征其形貌;利用动态光散射仪检测其粒径;利用近红外热成像仪表征其光热转换效能;考察药物的载药率及体外释放情况。体外的抗肿瘤实验中,将骨肉瘤MG-63细胞分为5组:PBS对照组、游离DOX组、Au@PDA+激光组、Au@PDA@DOX组和Au@PDA@DOX+激光组。采用CCK-8法及活死细胞染色检测各实验组对骨肉瘤MG-63细胞的杀伤效果,使用电感耦合等离子体质谱(ICP-MS)去检测骨肉瘤对Au@PDA@DOX和Au@PDA的摄取情况,使用荧光倒置显微镜观察各组细胞凋亡情况。结果:成功制备出粒径为(13.3±0.8)nm的纳米金粒子,使用聚多巴胺包裹的纳米金粒子粒径为(185.6±6.0)nm,负载DOX后粒径增大到了(288.0±6.2)nm,载药量为(12.8±0.8)%,在酸性条件下,DOX的释放速度加快;细胞吞噬实验表明MG-63细胞对Au@PDA@DOX纳米粒子的吞噬量高于Au@PDA纳米粒子;单一治疗及光热治疗与化疗协同杀伤MG-63细胞的能力评估结果表明,联合作用的效果大于单一治疗。结论:负载DOX的Au@PDA纳米系统表现出良好的光热效应,能够抑制MG-63细胞的生长。该纳米系统实现了化疗与光热协同作用,可用于骨肉瘤的治疗。

Objective:To explore the application effect of polydopamine modified gold nanoparticles loaded with doxorubicin(DOX)for chemo-photothermal synergistic therapy in osteosarcoma.Methods:Gold nanoparticles(Au),polydopamine modified gold nanoparticles(Au@PDA),and polydopamine modified gold nanoparticles loaded with DOX(Au@PDA@DOX)were prepared.The morphology and diameter of nanoparticles were characterized by high power transmission electron microscope(TEM)and dynamic light scatterometer(DLS),respectively.The photothermal conversion efficiency was detected by near infrared thermal imaging instrument,and the drug loading rate and release in vitro were also investigated.MG-63 cells were divided into five groups:PBS control group,free DOX group,Au@PDA+NIR(808 nm,1 W/cm^(2) near infrared laser)group,Au@PDA@DOX group and Au@PDA@DOX+NIR group.The inhibition effect of MG-63 cells in each experimental group was detected by CCK-8 and living/dead cell staining.The uptake of Au@PDA@DOX and Au@PDA by osteosarcoma cells was detected by inductively coupled plasma mass spectrometry(ICP-MS).The apoptosis of cells in each group was observed by fluorescence inverted microscope.Results:Gold nanoparticles with a diameter of(13.3±0.8)nm were prepared successfully.The particle size of gold nanoparticles coated with polydopamine was(185.6±6.0)nm.The particle size was increased to(288.0±6.2)nm,and drug loading rate was(12.8±0.8)%after loading DOX.Under acidic condition,DOX can be rapidly released from the surface of the material.Cell phagocytosis test showed that the phagocytosis amount of Au@PDA@DOX nanoparticles was higher than the Au@PDA nanoparticle.The chemo-photothermal synergistic therapy showed the better tumor inhibition effect on osteosarcoma cells.Conclusion:The Au@PDA nanoparticle loaded with DOX shows good photothermal effect and could inhibit the growth of MG-63 cells.The nanosystem realizes the synergistic effect of chemotherapy and photothermal therapy,and can be used in the treatment of osteosarcoma.