葡聚糖纳米凝胶与淋巴管内皮细胞相容性研究  

Experimental research on cell biocompatibility of dextran-based fluorescent nanogel acids with lymphatic endothelial cells

作　　者：崔春晓[1] 戴婷婷[1] 周淑彦[2] 蒋朝华[1] 窦红静[2] 周广东[1] 李圣利[1] 

机构地区：[1]上海交通大学医学院附属第九人民医院整复外科,200011 [2]上海交通大学材料科学与工程学院

出　　处：《中华实验外科杂志》2015年第9期2126-2129,共4页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金资助项目（81371700、81372080）;九院基金资助项目（2013B02）

摘　　要：目的 观察人淋巴管内皮细胞（LECs）与生物材料葡聚糖-聚丙烯酸荧光纳米凝胶（FDNG）的相容性.方法 采用荧光分光光度仪和纳米粒度电位分析仪测定葡聚糖-聚丙烯酸荧光纳米凝胶（FDNG）的表征,噻唑蓝（MTT）检测FDNG对LECs毒性及增殖影响;将LECs同FDNG共孵育,荧光显微镜检测其是否发生复合,并通过流式细胞仪检测其复合率;透射电镜检测FDNG在淋巴管内皮细胞内定位及代谢方式;通过流式细胞仪检测其表面标志物、人工基质胶（Matrigel）三维成管实验检测其对淋巴管内皮细胞表型和功能的影响.结果 FDNG粒径180 nm,在波长为492 nm的激发光下发出绿色荧光;MTT检测结果显示与不同浓度FDNG共孵育组的活细胞数量和增殖速度同对照组比较,差异无统计学意义（P＞0.05）;LEC和FDNG共孵育24 h后,99.4％的LECs摄取FDNG,荧光显微镜下发出绿色荧光,相差显微镜下呈典型的铺路石状;流式细胞学检测显示正常淋巴管内皮细胞和复合FDNG的淋巴管内皮细胞表面特异性标记表达率比较,差异无统计学意义（P＞0.05）.成管试验显示,吞噬了FDNG的LECs可以成管.结论 葡聚糖-聚丙烯酸荧光纳米凝胶具有良好的淋巴管内皮细胞相容性.Objective To investigate the cell biocompatibility of dextran-based fluorescent nanogel （FDNG） with lymphatic endothelial cells （LECs）.Methods Wavelength of excitation light of FDNG was measured with fluorescence spectrophotometer;Diameter of FDNG was examined using Zetasizer Nano ZS90.Methyl thiazol tetrazolium （MTT） test was performed for cytotoxicity of FDNG and evaluation of LECs proliferation after co-cultured with FDNG.Uptake of FDNG by LECs was observed under phase contrast microscope and fluorescence microscope after co-culturing LECs with FDNG for 24 hours,uptake rate was further evaluated by flow cytometry.The distribution of intracellular FDNG was positioned with Transmission electronic microscope.We compared lymphatic markers expression rate in LECs and FDNG-LECs by flow cytometry.FDNG-LECs were also seeded on Matrigel for tube formation assay.Results FDNG emitted green light excited by light of 492 nm.Diameter of FDNG was 180 nm.MTT results demonstrated no significant difference of proliferation speed and viable cells numbers could be observed between LECs co-cultured with FDNG group and control group （P ＞ 0.05）.Flow cytometry revealed that 99.4％ of LECs could take in FDNG and the uptake exert minimal influence on LEC morphology,phenotype and tube formatiou ability.TEM results suggested FDNG positioned in lysosome.Conclusion FDNG was well biocompatible with LECs,and could be used for further investigation for lymphatic imaging.

关 键 词：荧光纳米凝胶 葡聚糖 造影 淋巴管内皮细胞 生物安全性 

分 类 号：R735.370.2[医药卫生—肿瘤]