载10-羟基喜树碱液态氟碳靶向纳米粒的制备及体外双模态成像  

Preparation and in vitro dual-mode imaging of 10-HCPT-loaded liquid fluorocarbon nanoparticles

作　　者：程武松 陈洪波 游玉峰 CHENG Wusong;CHEN Hongbo;YOU Yufeng(Department of Urology,Central Hospital of Enshi Tujia and Miao Autonomous Prefecture,Hubei 445000,China)

机构地区：[1]恩施土家族苗族自治州中心医院泌尿外科,湖北省恩施市445000 [2]恩施土家族苗族自治州中心医院放射科,湖北省恩施市445000

出　　处：《临床超声医学杂志》2020年第9期641-646,共6页Journal of Clinical Ultrasound in Medicine

基　　金：国家自然科学基金面上项目(81660291)。

摘　　要：目的制备载10-羟基喜树碱(10-HCPT)液态氟碳靶向纳米粒,观察其对人卵巢癌SKOV-3细胞的寻靶能力及体外超声/CT双模态成像效果。方法以羟基端乳酸/羟基乙酸共聚物(PLGA-COOH)、全氟溴辛烷(PFOB)及10-HCPT为原料,采用双乳化法制备PLGA@10-HCPT-PFOB纳米粒,观察其外部形态和内部结构,测量粒径大小和表面电位,以及10-HCPT的包封率和载药量;通过碳二亚胺法连接cRGD肽制备cRGD-PLGA@10-HCPT-PFOB靶向纳米粒,使用共聚焦显微镜和流式细胞仪检测PLGA@10-HCPT-PFOB纳米粒与cRGD肽的连接情况。体外实验检测其对人卵巢癌SKOV-3细胞的靶向性能;观察靶向纳米粒的体外超声/CT成像能力。结果本实验制得的靶向载药纳米粒外观呈黄色混悬液,光镜下纳米粒形态规则、大小均一,平均粒径(322.03±5.34)nm,平均表面电位(-1.55±0.10)mV;扫描电镜示靶向纳米粒呈球形,表面光滑;透射电镜示其为核壳结构,PLGA包裹PFOB和10-HCPT,10-HCPT包封率和载药量分别为(81.34±2.28)%和(13.24±1.24)%;共聚焦显微镜观察示FITC标记的cRGD肽呈绿色荧光,与DiI标记染色的红色纳米粒共同融合呈橙黄色荧光,流式细胞仪检测PLGA@10-HCPT-PFOB纳米粒FITC荧光强度为0.78%,cRGD-PLGA@10-HCPT-PFOB纳米粒FITC荧光强度为80.76%;共聚焦显微镜及流式细胞仪显示大量纳米粒靶向到SKOV-3细胞表面,与细胞膜上的αvβ3受体结合,部分被SKOV-3细胞吞噬;随着靶向纳米粒的浓度不断增加,其体外超声/CT成像明显增强。结论本实验成功制备载10-HCPT液态氟碳靶向纳米粒,其10-HCPT包封率和载药量均较高,对人卵巢癌SKOV-3细胞有较强的靶向性,通过聚集显影可增强体外超声/CT成像。Objective To prepare 10-hydroxycamptothecin(10-HCPT)loaded liquid fluorocarbon targeting nanoparticles,and to investigate their targeting ability to ovarian carcinoma cells(SKOV-3)and the effect of ultrasound/computed tomography(CT)dual-mode imaging in vitro.Methods By using hydroxy terminated lactic acid/glycolic acid copolymer(PLGA-COOH),perflurooctyl bromide(PFOB)and 10-HCPT,PLGA@10-HCPT-PFOB nanoparticles were prepared by double emulsion solvent evaporation method.The surface morphology and internal structure were observed,the particle size,surface potential,encapsulation efficiency and drug loading of 10-HCPT were measured as well.The cRGD peptide was conjugated to the nanoparticles by carbodiimide technique.The binding of cRGD-PLGA@10-HCPT-PFOB was detected by laser scanning confocal microscope and flow cytometer.The targeting properties of the nanoparticles to SKOV-3 were detected.The ability of ultrasound/CT imaging of targeted nanoparticles was evaluated in vitro.Results The targeted nanoparticles were produced as a yellow suspension.The size distribution of the nanoparticles was uniform observed by optical microscope,with a mean diameter of(322.03±5.34)nm and surface potential was(-1.55±0.10)mV.The surface of the targeted nanoparticles was smooth observed by the scanning electron microscope.The core-shell structure was observed by the transmission electron microscope,and the PFOB and 10-HCPT were trapped in PLGA.The drug encapsulation and drug loading capacity of 10-HCPT were(81.34±2.28)%and(13.24±1.24)%,respectively.Laser scanning confocal microscope showed that the FITC-labeled cRGD peptides were green and it fused to orange with the red nanoparticles labeled DiI.The flow cytometer showed that the intensity of FITC in PLGA@10-HCPT-PFOB and cRGD-PLGA@10-HCPT-PFOB nanoparticles were 0.78%and 80.76%,respectively.The laser scanning confocal microscope and flow cytometry showed that a large number of nanoparticles were targeted to SKOV-3 cells,binding toαvβ3 receptor,some of which were uptaken by SKO

关 键 词：10-羟基喜树碱 靶向 纳米粒 成像 体外 

分 类 号：R445.1[医药卫生—影像医学与核医学]