| 宣吉晴,陈瑜莉,敖梦,王志刚,郑元义,汪朝霞,李奥.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[J].中国医学影像技术,2017,33(6):810~815 |
| 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 |
| Experimental study on preparation and targeting research in vitro of targeted nanoparticle ultrasound contrast agent carrying cRGD |
| 投稿时间：2017-02-15 修订日期：2017-04-27 |
| DOI：10.13929/j.1003-3289.201702043 |
| 中文关键词: 靶向 纳米粒 超声检查 造影剂 |
| 英文关键词:Target Nanoparticles Ultrasonography Contrast media |
| 基金项目: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）。 |
| 作者 | 单位 | E-mail | | 宣吉晴 | 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 400010 | | | 陈瑜莉 | 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 400010 | | | 敖梦 | 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 400010
重庆医科大学附属第二医院超声科, 重庆 400010 | aomeng735@163.com | | 王志刚 | 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 400010
重庆医科大学附属第二医院超声科, 重庆 400010 | | | 郑元义 | 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 400010
重庆医科大学附属第二医院超声科, 重庆 400010
上海交通大学附属第六人民医院超声科, 上海 200233 | | | 汪朝霞 | 重庆医科大学附属儿童医院超声科, 重庆 400016 | | | 李奥 | 南京医科大学第一附属医院超声科, 江苏 南京 210029 | |
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| 中文摘要: |
| 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PLGA-PFOB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。 |
| 英文摘要: |
| Objective To prepare a poly (lactic-co-glycolic acid) (PLGA) nanoparticles ultrasound contrast agent carrying 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, distribution and Zeta potential were observed using Malvern particle size analyzer. The cRGD was conjugated onto the nanoparticles 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)mV. 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 cRGD 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 nanoparticles carrying cRGD (cRGD-NPs) can be prepared successfully. The cRGD-NPs contrast agent has strongly specific affinity on the HSC. |
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