靶向抑制晶状体上皮细胞增殖的载药毫微球的研究  

STUDY OF DRUG-LOADED IMMUNONANOPARTICLES TARGETING TO LENS EPITHELIAL CELLS

作　　者：刘晓燕[1] 常津[1] 郭艳霜[1] 原续波[1] 刘新玲 李筱荣[2] 

机构地区：[1]天津大学材料学院纳米生物技术研究所,天津300072 [2]天津医科大学眼科中心,天津300070

出　　处：《中国生物医学工程学报》2004年第6期583-588,共6页Chinese Journal of Biomedical Engineering

基　　金：国家自然科学基金 (No .5 0 3 73 0 3 3 ) ;天津市自然科学基金 (No .0 0 3 70 2 711)

摘　　要：采用碳二亚胺法使抗人晶状体上皮细胞单克隆抗体与聚乳酸载 5 -氟尿嘧啶毫微球偶联 ,制备出抗人晶状体免疫毫微球。以牛血清白蛋白作乳化剂采用复乳法制备聚乳酸载 5 -氟尿嘧啶毫微球 ,使毫微球表面吸附牛血清白蛋白 ,再将抗人晶状体上皮细胞单克隆抗体与聚乳酸载 5 -氟尿嘧啶毫微球偶联。采用扫描电镜和透射电镜观察微球形貌和结构 ,用动态光散射粒径分析仪测载药毫微球粒径。ELISA法检测偶联后的抗体活性 ,MTT法检测免疫毫微球对晶状体上皮细胞的抑制作用 ,间接免疫荧光法检测该免疫毫微球与晶状体上皮细胞的特异性结合能力和细胞内化过程。结果显示载药毫微球表面光滑 ,平均粒径为 191.0± 0 .2 0 2nm ,其载药率为 8.2 % ,药物利用率为2 4 .6 %。免疫载药毫微球中的单克隆抗体HILE6保留达到原免疫活性 84 % ,2 4h对兔晶状体上皮细胞抑制率可达6 8.4 2 %。该免疫载药毫微球能与晶状体上皮细胞特异性结合 ,30min后可吸附到晶状体上皮细胞上 ,在 4h内进入细胞质最后进入细胞核。该实验为特异性抑制晶状体上皮细胞增殖 ,预防白内障术后并发症 -后囊混浊提供了重要的科学依据。The purpose of this work is to prepare the immunonanoparticles [HDLE6 - PLA(5 - Fu) - NPs] (INPs) by cross - linking the monoclonal antibody HDLE6 to the PLA nanoparticles containing 5 - fluorouracil(5 - Fu) [PLA(5 - Fu) - NPs]. The NPs were characterized for morphology and structure by scanning electronic microscopy (SEM) and transmission electronic microscopy (TEM). The particle sizes of NPs were determined by Particle Size Analyzer, 5 - FU drug content of NPs wer also examined. ELISA method was used to evaluate the immunological activity of antibody, the killing effects of the INPs on lens epithelial cells (LECs) were evaluated by MTT assay. The indirect immunofluorescence method was used for evaluating the selectivity of the INPs to LECs and the intemalization of the INPs. The results showed that NPs had smooth external morphology, the average size was approximately 191.0 ± 0.202nm, and the drug loading was 8.2%. The immunological activity of antibody could reach 84% of the original HILE6, 68.42% growth inhibition of LECs was achieved with 240μg/ml of INPs after 24 hours of exposure. The INPs could selectively adhere the lens epithelial cells in 30min and enter the cytoplasm or/and nucleus in 4h, and the nucleus were the major compartment of their distribution. The result is of significance for selectively and long - time killed LECs for preventing posterior capsular opacification.

关 键 词：免疫毫微球 聚乳酸毫微球 抗人晶状体上皮细胞单克隆抗体 晶状体上皮细胞 5-氟尿嘧啶 

分 类 号：R318.08[医药卫生—生物医学工程]