相变型介孔硅纳米探针超声/近红外双模态显像及体外溶栓的实验研究  

Experimental study on ultrasound/near-infrared dual-mode imaging and thrombolysis in vitro of phase-change mesoporous silicon nanoprobe

作　　者：王治文 姜楠[1] 潘桔红 曹省[1] 江智欣 周青[1] 胡波[1] Wang Zhiwen;Jiang Nan;Pan Juhong;Cao Sheng;Jiang Zhixin;Zhou Qing;Hu Bo(Department of Ultrasound Imaging,Renmin Hospital of Wuhan University,Wuhan 430060,China)

机构地区：[1]武汉大学人民医院超声影像科,武汉430060

出　　处：《中华超声影像学杂志》2023年第10期907-914,共8页Chinese Journal of Ultrasonography

基　　金：国家自然科学基金(82102045, 81901757)。

摘　　要：目的探讨靶向血小板诊疗一体化的纳米探针(RGD/ICG/PFP@MSN)在体外的超声/近红外双模态显像潜能及对血栓的协同治疗作用。方法采用超声声振及碳二亚胺法制备以介孔二氧化硅纳米粒(MSN)为载体, 负载精氨酸-甘氨酸-天冬氨酸序列肽(RGD)、全氟戊烷(PFP)及吲哚箐绿(ICG)的纳米探针(RGD/ICG/PFP@MSN)。用扫描和透射电镜观察探针形态;用蛋白定量法(BCA)和紫外-可见光-近红外光谱分别检测RGD、ICG的载入;体外研究纳米探针在近红外光(NIR)照射下的成像性能、光热响应能力;用细胞毒性实验和溶血实验检测其生物安全性;在超声及NIR辐照下研究其相变情况;将探针与新鲜动脉血栓孵育后行冰冻切片观察其寻靶能力, 并行超声和NIR辐照治疗, 以辐照前后血栓重量变化评估其溶栓能力。结果制备的纳米探针形态规则, 大小均匀, 粒径(156.83±5.05)nm, 电位(11.47±0.25)mV。RGD偶联率为(77.67±4.50)%, 能介导纳米探针靶向体外新鲜动脉血栓;紫外-可见光-近红外光谱证实了ICG的成功载入, 其包封率为(80.47±0.05)%。超声和NIR辐照后纳米探针能发生声致相变和热致相变并增强超声显影效果;随着纳米探针溶液浓度的增加, NIR成像信号逐渐增强, 且经NIR照射后呈浓度依赖性和辐照强度依赖性升温。细胞毒性实验和溶血实验显示该探针具有良好的生物安全性, 且探针在联合超声与NIR的辐照下能发挥溶栓作用, 血栓重量经治疗后显著减少(P<0.01)。结论构建的RGD/ICG/PFP@MSN纳米探针具备优良的超声与NIR双模态成像能力、良好的相变能力和光热转换效力, 以及针对血栓的高效靶向渗透和治疗作用, 可为实现在超声与NIR协同作用下对血栓类疾病的诊疗一体化提供有力的体外实验证据和新策略。Objective To explore the potential of thrombus-targeted nanoprobes for ultrasound/near-infrared bimodal imaging and their synergistic therapeutic effects on thrombosis in vitro.Methods Nanoprobes loaded with arginine-glycine-aspartate peptide(RGD),perfluoropentane(PFP)and indocyanine green(ICG)were prepared by ultrasonic vibration and carbodiimide method with mesoporous silica nanoparticle(MSN)as the carrier.The probe morphology was observed by scanning and transmission electron microscopy.The loading of RGD and ICG was detected by Bicinchoninic Acid Assay(BCA)and UV-Visible-NIR spectroscopy respectively.The imaging performance and photothermal response of the nanoprobe under near infrared light(NIR)irradiation were studied in vitro.Its biological safety was tested by cytotoxicity test and hemolysis test.The phase transformation was studied under ultrasound and NIR irradiation.The nanoprobe was incubated with fresh arterial thrombus,and its target-seeking ability was observed by frozen section.Ultrasound and NIR irradiation were used to evaluate its thrombolytic ability by the weight changes of thrombus before and after irradiation.Results The prepared nanoprobe had regular morphology and uniform size.The particle diameter was(156.83±5.05)nm,and the surface potential was(11.47±0.25)mV.The RGD coupling rate was(77.67±4.50)%,which could mediate the targeting of nanoprobe to fresh extracorporeal arterial thrombus.UV-Visible-NIR spectroscopy confirmed the successful loading of ICG,and its encapsulation rate was(80.47±0.05)%.After ultrasound and NIR irradiation,the nanoprobe could undergo acoustically induced phase transition,thermally induced phase transition and enhance the ultrasonic development effect.With the increase of the concentration of the nanoprobe solution,the NIR signal gradually increased,and the temperature rose in a concentration-dependent and intensity-dependent manner after NIR irradiation.The cytotoxicity test and hemolysis test showed that the nanoprobe had good biological safety,and it could

关 键 词：超声检查 纳米探针 近红外 血栓 溶栓治疗 

分 类 号：R445.1[医药卫生—影像医学与核医学]