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。
Objective To prepare a poly (lactic-co-glycolic acid) (PLGA) nanoparticles ultrasound contrast agent carry ing cRGD, and to investigate its targeting ability to the rat hepatic stellate cells (HSC) in vitro. Methods PLGA-PFOB nanoparticles were prepared by a double emulsion solvent evaporation process with a modified polymeric shell (PLGA- COOH) encapsulating perflurooctyl bromide (PFOB). The morphological and structural characteristics of the nanoparticles were observed by an optical microscope, scanning electron microscopy and transmission electron microscopy. The size, dis- tribution and Zeta potential were observed using Malvern particle size analyzer. The cRGD was conjugated onto the nanop- articles by carbodiimide technique. The combination of cRGD with nanoparticles (cRGD-NPs) and the targeting function of cRGD-NPs in vitro were proved by laser scanning confocal microscopy (LSCM). Results The samples were milk white in deionized water, with a mean diameter of (255.3±66.8)nm and a polydispersity index of 0. 025. Zeta potential was (-16.4 ±5.1)inV. Scanning electron microscopy images reveal that majority of capsules were spherical with smooth surface. Transmission electron microscope resented spherical particle with a core-shell structure. By observing LSCM, Nile red- dyed NPs emitted red fluorescence, FITC-labeled eRGD emitted green fluorescence, and overlay fluorescence image emitted bright homogeneous yellow fluorescence. It was found that greater and stronger red fluorescence representing cRGD-NPs could be observed in the cytoplasm of HSC while fewer NPs remained within HSCs in the non-targeted group. Conlusion The PLGA encapsulated PFOB nanopartieles carrying cRGD (cRGD-NPs) can be prepared successfully. The cRGD-NPs contrast agent has strongly specific affinity on the HSC.
作者
宣吉晴
陈瑜莉
敖梦
王志刚
郑元义
汪朝霞
李奥
XUAN Jiqing;CHEN Yuli;AO Meng;WANG Zhigang;ZHENG Yuanyi;WANG Zhaoxia;LI Ao(Chongqing Key Laboratory of Ultrasound Molecular Imaging,the Second Affiliated Hospital of Chongqing Medical University,Chongqing 400010,China;Department of Ultrasound,the Second Affiliated Hospital of Chongqing Medical University,Chongqing 400010,China;Department of Ultrasound,Shanghai Jiao Tong University Affiliated Shanghai Sixth People’s Hospital,Shanghai 200233,China;Department of Ultrasound,the Children’s Hospital of Chongqing Medical University,Chongqing 400016,China;Department of Ultrasound,the First Affiliated Hospital of Nanjing Medical University,Nanjing 210029,China)
出处
《中国医学影像技术》
CSCD
北大核心
2017年第6期810-815,共6页
Chinese Journal of Medical Imaging Technology
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)
关键词
靶向
纳米粒
超声检查
造影剂
Target
Nanoparticles
Ultrasonography
Contrast media