基于中空复合纳米粒子的超声溶栓体系构建及效果验证  

Construction and Effect Verification of Ultrasonic Thrombolysis System Based on Hollow Composite Nanoparticles

作　　者：孙海悦 韦佼君 陈家乐 赵龙 Sun Haiyue;Wei Jiaojun;Chen Jiale;Zhao Long(School of Basic Medical Sciences,Chengdu Medical School,Chengdu 610500,China;School of Biological Sciences and Technology,Chengdu Medical College,Chengdu 610500,China)

机构地区：[1]成都医学院基础医学院,成都610500 [2]成都医学院生物科学与技术学院,成都610500

出　　处：《成都医学院学报》2024年第3期395-400,共6页Journal of Chengdu Medical College

基　　金：四川省中央引导地方科技发展专项项目(No:2023ZYD0082)。

摘　　要：目的 评价超声辅助微泡溶栓联合声动力疗法的中空复合纳米粒子的生物安全性及体外溶栓效果,评估其治疗静脉血栓栓塞症的潜在应用价值。方法 制备氮掺杂石墨烯量子点(NGQD)和一氧化氮供体N,N’-二仲丁基-N,N’-二亚硝基-1,4-苯二胺(BNN6),BNN6负载于NGQD上,将二者复合体载入利用盐模板法制备的中空介孔二氧化硅(HMSN)中,形成纳米溶栓体系BNM;通过电镜和纳米粒度仪观察BNM的形貌、尺寸等特征;通过细胞毒性实验、活-死细胞染色及溶血实验评价纳米溶栓体系BNM的生物安全性;通过体外溶栓和纤维蛋白凝块降解评价纳米材料的体外溶栓效果。结果 与对照组相比,不同浓度的BNM对HUVECs和RAW264.7细胞的活性无影响,中低浓度BNM的溶血率未超过纳米材料溶血安全限,3个浓度的BNM对红细胞和血小板的形貌未产生明显影响,不引起血小板活化;与对照组相比,超声辅助BNM组的溶栓率随BNM浓度增加而提高,纤维蛋白网络出现空洞,荧光面积比降低。结论 本研究制备的纳米溶栓系统具有很好的生物相容性和血液安全性,可以响应超声进行溶栓,表现出优良的溶栓性能,为临床应用提供了新策略。Objective To evaluate the biosafety and in vitro thrombolytic effect of hollow composite nanoparticles in ultrasound-assisted microbubble thrombolysis combined with sonodynamic therapy,and evaluate their potential in the treatment of venous thrombosis embolism(VTE).Methods Nitrogen-doped graphene quantum dots(NGQD)and N,N′-Di-sec-butyl-N,N′-dinitroso-1,4-phenylene diamine(BNN6)were prepared by referring to the methods in the literature.BNN6 was loaded on NGQD.And the composite was loaded on hollow mesoporous silica nanoparticles(HMSN)prepared by salt template method.Thus,a nanoparticle thrombolytic system BNN6/NGQD@hMSN(BNM)was developed.The morphology and size of BNM were observed by electron microscopy and nano particle size analyzer.The biosafety of BNM was evaluated by cytotoxicity test,live-dead cell staining and hemolysis test.The in vitro thrombolytic effect of BNM was evaluated by in vitro thrombolysis and fibrin clot degradation.Results Compared with the control group,different concentrations of BNM had no effect on the activity of human umbilical vein endothelial cells(HUVECs)and mouse mononuclear macrophages RAW264.7 cells,the hemolysis rate of medium and low concentrations of BNM did not exceed the hemolysis safety limit of nanomaterials,three concentrations of BNM had no significant effect on the morphology of red blood cells and platelets,and did not cause platelet activation.Compared with the control group,the thrombolysis rate in the ultrasound-assisted BNM group increased with the increase of BNM concentration,the fibrin network showed voids,and the fluorescence area ratio decreased.Conclusion The nano-thrombolysis system prepared in this study has good biocompatibility and blood safety,and can perform thrombolysis in response to ultrasound,showing excellent thrombolysis performance,and providing a new strategy for clinical application.

关 键 词：超声溶栓 声动力疗法 石墨烯量子点 一氧化氮供体 中空介孔二氧化硅纳米颗粒 

分 类 号：R318.08[医药卫生—生物医学工程]