包裹吲哚菁绿的靶向相变型阳离子脂质纳米粒的制备及体外显像  

Preparation of targeted cationic nanoparticles incoperated with indocyanine green and imaging with the particles in vitro

作　　者：邹宏密 黎梦 周希瑗[1] 王志刚[2] 简嘉[1] ZOU Hongmi,LI Meng,ZHOU Xiyuan,WANG Zhigang,JIAN Jia(Department of Ophthalmology,the Second Hospital Affiliated to Chongqing Medical University, Chongqing 400010, China)

机构地区：[1]重庆医科大学附属第二医院眼科,重庆市400010 [2]重庆医科大学超声影像学研究所

出　　处：《临床超声医学杂志》2018年第6期361-365,共5页Journal of Clinical Ultrasound in Medicine

基　　金：国家自然科学基金项目(81170858;81501487)

摘　　要：目的制备一种包裹吲哚菁绿(ICG)的靶向相变型阳离子脂质纳米粒(INP),连接CD105抗体,检测其光热效应、体外相变、光声及超声显像规律。方法采用双乳化法制备包裹ICG及液态氟碳的INP;链霉亲和素法连接CD105抗体,流式细胞仪及激光共聚焦显微镜下观察INP与抗体结合情况;脉冲激光激发观察其光热效应及相变情况;体外低强度聚焦超声辐照,记录各时间点超声显影效果;光声成像仪检测其光声显像能力。结果制备的INP平均粒径(354.20±93.85)nm,平均电位(25.20±3.29)m V,测得ICG包封率为(89.56±0.79)%;与CD105抗体结合率为99.89%。纳米粒具有良好光热效应,其中在浓度5 mg/ml、功率2 W/cm2激光辐照180 s时其温度可升至63℃;可发生液气相变。体外低强度聚焦超声于3 W功率作用0.5、2.5、5.0、7.5 min,B-mode模式及Contrast模式均显示5 min时回声强度升至最高。光声信号随纳米粒浓度升高而增强,具有良好的线性关系。结论成功制备了包裹ICG的靶向相变型阳离子INP,其与CD105抗体结合率高,且光热效应好,光声及超声显影效果均佳。Objective To prepare a targeted phase change cationic lipid nanoparticle（INP）coated with indocyanine green（ICG）,and CD105 antibody was attached to the particles.The photothermal effect,phase change,imaging laws of photoacoustic and ultrasonic imaging in vitro were studied.Methods INP coated with ICG and liquid fluorocarbon（PFP）was prepared by double emulsification method.Streptavidin method was used to connect CD105 antibody.Flow cytometry and laser confocal microscopy were used to observe antibody binding.The photothermal effect and phase change were observed by pulsed laser excitation.The effect of ultrasound imaging was recorded at different time after the phase change caused by low intensity focused ultrasound irradiation in vitro,and the photoacoustic imaging ability was detected by photoacoustic imager.Results The average size of the prepared INP was（354.20±93.85）nm,the average potential was（25.20±3.29）m V,the entrapment rate was（89.56±0.79）%,and the binding rate of CD105 antibody was 99.89%.The nanoparticles have good photothermal effect,and the temperature can rise to 63℃ with laser irradiation at a concentration of 5 mg/ml and a power of 2 W/cm^2 for 180 s,and liquid-gas phase change occurs.The echo intensity increased to the highest when irradiated with low intensity focused ultrasound with contrast mode for 5 min,and with B-mode at a power of 3 W for 0.5,2.5,5.0,and 7.5 min,respectively.The echo intensity increased with the increase of nanoparticle concentration,there was a good linear relationship between nanoparticle concentration and echo intensity.Conclusion The target phase variant cationic INP coated with ICG were successfully prepared,with good characteristics of light-thermal effect,photoacoustic imaging and ultrasound imaging,and high binding rate of nanoparticles to CD105 antibody.

关 键 词：阳离子 脂质纳米粒 吲哚菁绿 相变 靶向 

分 类 号：R445.1[医药卫生—影像医学与核医学]