| 摘要: |
| 目的 制备载10-羟基喜树碱(10-hydroxycamptothecin, 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(perflurooctyl bromide,PFOB)和10-HCPT为原料,采用成熟的双乳化法制备PLGA@-HCPT-PFOB纳米粒,采用光学显微镜、扫描电镜以及透射电镜观察其外部形态以及内部结构；马尔文粒径分析仪测量其粒径大小及表面电位；紫外分光光度测定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,用激光共聚焦显微镜检测PLGA@10-HCPT-PFOB与cRGD肽的连接情况,在体外实验检测其对卵巢癌SKOV-3细胞的靶向性能；观察靶向纳米粒的体外超声成像和CT成像能力。结果 所制得的靶向载药纳米微球为黄色混悬液,光镜下纳米粒大小、分布均匀,马尔文粒径分析仪测量其平均粒径为(322.03±5.34)nm,Zeta电位为(-1.55±0.10)mV；扫描电镜显示该靶向纳米粒呈球形、表面光滑,透射电镜示其为核壳结构,PLGA包裹PFOB和10-HCPT,其10-HCPT包封率和载药量分别为(81.34±2.28)%和(13.24±1.24)%；激光共聚焦显微镜观察到标记FITC的cRGD肽显示为绿色,与标记DiI的红色纳米粒共同融合成橙黄色；激光共聚焦显微镜及流式细胞仪显示大量纳米粒靶向到细胞表面,与细胞膜上的ανβ3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,其体外超声及CT成像明显增强。结论 成功制备载10-HCPT液态氟碳靶向纳米粒,该纳米粒10-HCPT包封率、载药量较高,对卵巢癌SKOV-3细胞有较强的靶向性,通过聚集显影,可增强体外超声及CT成像。 |
| 关键词: 10-羟基喜树碱 靶向 纳米粒 成像 |
| DOI： |
| 投稿时间：2020-03-17修订日期：2020-04-29 |
| 基金项目:]国家自然科学基金面上项目(81660291)。 |
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| Preparation and in vitro dual-mode images of 10-HCPT-loaded liquid fluorocarbon nanoparticles |
| Cheng Wusong,You Yufeng,Chen Hongbo |
| (Department of Urology,the Central Hospital of Enshi Tujia and Miao Autonomous Prefecture Enshi) |
| Abstract: |
| ［］Objective To prepare 10-hydroxycamptothecin(10-HCPT) loaded liquid fluorocarbon target nanoparticles, and to investigate its targeting ability to ovarian carcinoma cells (SKOV-3) and the effect of Ultrasound/CT dual-mode imaging in vitro. Methods PLGA@10-HCPT-PFOB nanoparticles were prepared by double emulsion solvent evaporation process with modified polymeric shell (PLGA-COOH) encapsulating perflurooctyl bromide (PFOB) and 10-HCPT, its general physical properties were observed with optical microscope (OM), scanning electron microscope (SEM) and transmission electron microscope (TEM). The particle size, distribution and Zeta potential were measured by Marvin particle size analyzer. The encapsulation rate and drug load of 10-HCPT were measured by ultraviolet spectrophotometer. The cRGD peptide was conjugated to the nanoparticles by carbodiimide technique. The binding of PLGA@10-HCPT-PFOB to cRGD peptide was detected by Laser Scanning Confocal Microscope (LSCM), and the targeting properties of the nanoparticles were detected in ovarian cancer cells (SKOV-3).The ability of ultrasound imaging and computed tomography(CT) imaging was evaluated in vitro. Results The prepared targeted nanoparticles were yellow suspension, the size distribution of the nanoparticles was uniform under optical microscope, the surface of the targeted nanoparticles was smooth under the scanning electron microscope, the core-shell structure was shown under the transmission electron microscope, and the PFOB and 10-HCPT were wrapped in PLGA, with mean diameter of(322.03±5.34)nm and Zeta potential was(-1.55±0.10)mV. The drug encapsulation and drug loading capacity of cRGD-PLGA@10-HCPT- |
| Key words: 10-hydroxycamptothecin Target Nanoparticles Imaging |
