携带cRGD肽的靶向纳米粒超声造影剂的制备以及体外寻靶实验研究

发布时间：2018-06-12 06:31

本文选题：靶向 + 纳米粒　；参考：《中国医学影像技术》2017年06期

【摘要】：目的制备携带精-甘-天冬氨酸(cRGD)肽的聚乳酸/羟基乙酸(PLGA)纳米粒超声造影剂,观察其在体外对大鼠肝星状细胞(HSC)的靶向能力。方法以PLGA-COOH和全氟新溴烷(PFOB)为原料,采用双步乳化法制备PLGAPFOB纳米粒,采用光学显微镜、扫描电镜以及透射电镜观察其表面以及内部结构;采用马尔文粒径分析仪测量其粒径大小、分布以及表面电位;通过碳二亚胺法连接cRGD肽制备靶向纳米粒超声造影剂(cRGD-NPs),用激光共聚焦显微镜以及流式细胞仪检测PLGA-PFOB纳米粒造影剂与cRGD肽的连接情况;在大鼠HSC中验证该造影剂的靶向性能。结果所得样品为乳白色混悬液,纳米粒大小均匀,粒径分布较窄,平均粒径(255.3±66.8)nm,多分散指数为0.025,Zeta电位为(-16.4±5.1)mV;扫描电镜示纳米粒表面光滑规则、透射电镜示PLGA外壳里包裹液氮氟碳PFOB;共聚焦显微镜观察到连接FITC-cRGD显示为绿色,与显示红色的纳米粒共同融合成橙色,靶向纳米粒超声造影剂大量向大鼠HSC聚集并被其吞噬,红色荧光强度明显多于普通非靶向造影剂。结论成功制备的携带cRGD肽的纳米粒超声造影剂,在体外实验中对大鼠HSC有较强的特异性亲和力。
[Abstract]:Objective to prepare polylactic acid / hydroxyacetic acid (PLGA) nanoparticle ultrasound contrast agent carrying cRGD peptide and observe its targeting ability to rat hepatic stellate cells (HSC) in vitro. Methods using PLGA-COOH and perfluoroalkanane (PFOB) as raw materials, PLGAPFOB nanoparticles were prepared by double step emulsion method, and optical microscopy and scanning electron microscopy were used. The size, distribution and surface potential of the particle size, distribution and surface potential were measured by the Malvin particle size analyzer. The target nanoparticle ultrasound contrast agent (cRGD-NPs) was prepared by connecting cRGD peptide with the carbon two imide method. The PLGA-PFOB nanoparticle contrast agent and cRGD were detected by laser confocal microscope and flow cytometry. The targeting performance of the peptide was verified in the rat HSC. The results obtained were milky white suspension, the size of the nanoparticles was uniform, the particle size was narrow, the average particle size (255.3 + 66.8) nm, the polydispersity index was 0.025, the Zeta potential was (-16.4 + 5.1) mV; the scanning electron microscope showed the smooth rule of the nanoparticles surface, and the transmission electron microscope showed the PLGA shell. The fluorocarbon PFOB was encapsulated in liquid nitrogen; confocal microscopy showed that the connection FITC-cRGD was green and fused with the red nanoparticles into orange color. The target nanorage ultrasound contrast agents were aggregated and phagocytic in a large number of rat HSC, and the red fluorescence intensity was more than that of the ordinary non target contrast agent. The ultrasound contrast agent has a strong affinity for rat HSC in vitro.
【作者单位】：重庆医科大学附属第二医院超声分子影像学重庆市重点医学实验室;重庆医科大学附属第二医院超声科;上海交通大学附属第六人民医院超声科;重庆医科大学附属儿童医院超声科;南京医科大学第一附属医院超声科;
【基金】：国家自然科学基金青年基金(81501482、81401427);国家自然科学基金(81227801) 国家杰出青年科学基金(81425014)
【分类号】：R445.1

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