CAJ | 학술논문

目的制备一种新型血管外靶向超声造影剂——包裹液态氟碳的人表皮生长因子受体2(HER2)靶向高分子纳米球,并观察其体外寻靶、显影能力.方法首先采用单乳化法制备包裹液态氟碳的高分子PLGA纳米球(PFH-PLGA),然后通过碳二亚胺法将靶向HER2的特异性抗体trastuzumab连接到纳米球表面,制成靶向纳米球(PFH-PLGA-tra);检测其一般表征;免疫荧光法检测trastuzumab与纳米球连接情况;体外寻靶实验观察PFH-PLGA-tra与人乳腺癌细胞MCF-7的结合情况;体外加热法观察液态氟碳相变情况;体外观察纳米球增强超声显影效果.结果靶向液态氟碳造影剂平均粒径为(261±28)nm;免疫荧光法显示trastuzumab被成功地连接到了纳米球上;体外寻靶实验示,该靶向造影剂较多并牢固地聚集在表达HER2的MCF-7细胞表面,对照组未见造影剂和MCF-7的结合;体外热相变实验示,较多纳米球转变为微泡;体外显影实验示,造影剂回声强度和灰度均较双蒸水明显增强(P＜0.05).结论通过单乳化法和碳二亚胺法成功制备了靶向HER2的液态氟碳高分子超声造影剂,其粒径较小,靶向能力较强,具有相变潜能,有望成为HER2阳性肿瘤血管外显影、治疗的理想造影剂.
목적 제비일충신형혈관외파향초성조영제——포과액태불탄적인표피생장인자수체2(HER2)파향고분자납미구,병관찰기체외심파、현영능력.방법 수선채용단유화법제비포과액태불탄적고분자PLGA납미구(PFH-PLGA),연후통과탄이아알법장파향HER2적특이성항체trastuzumab련접도납미구표면,제성파향납미구(PFH-PLGA-tra);검측기일반표정;면역형광법검측trastuzumab여납미구련접정황;체외심파실험관찰PFH-PLGA-tra여인유선암세포MCF-7적결합정황;체외가열법관찰액태불탄상변정황;체외관찰납미구증강초성현영효과.결과 파향액태불탄조영제평균립경위(261±28)nm;면역형광법현시trastuzumab피성공지련접도료납미구상;체외심파실험시,해파향조영제교다병뢰고지취집재표체HER2적MCF-7세포표면,대조조미견조영제화MCF-7적결합;체외열상변실험시,교다납미구전변위미포;체외현영실험시,조영제회성강도화회도균교쌍증수명현증강(P＜0.05).결론 통과단유화법화탄이아알법성공제비료파향HER2적액태불탄고분자초성조영제,기립경교소,파향능력교강,구유상변잠능,유망성위HER2양성종류혈관외현영、치료적이상조영제.
Objective To prepare a novel extravascular human epithelial growth factor receptor 2 (HER2)-targeted polylactic-co-glycolic acid (PLGA)-COOH ultrasound contrast agent with liquid perfluorocarbons(PFH-PLGA-tra),and to observe it's general properties and it's effects on ultrasound imaging in vitro.Methods Polymeric nanocapsules of liquid perfluorocarbons were prepared by the single emulsion technique and conjugated with trastuzumab monoclonal antibody by EDC/NHS.The general characteristics were observed.The conjugation was demonstrated by immunofluorescence and the targeting performance of the agent was checked in human MCF-7 cells line in vitro.The encapsulation of liquid fluorocarbons was detected by heating in vitro.The effects on ultrasound imaging were observed in vitro.Results The mean diameter of agents was (261 ± 28)nm.The targeted nanocapsules were positive in immunofluorescence.In the targeted study,it was shown that a number of targeted nanocapsules conjugated with MCF-7 cells in vitro.Droplet-to-bubble transition experiment in vitro showed that there were no nano/microbubbles formed by heating in control groups at 80℃,while there were a lot of nano/microbubbles appeared in nanocapsules formulations at 80 ℃.The agents presented a good ultrasound imaging in vitro.Conclusions The HER2-targeted polymeric nanocapsules of liquid perfluorocarbons are successfully prepared,which may become a novel extravascular targeted ultrasound contrast agents.