载神经生长因子纳米药物体外诱导PC12细胞的药效  

Pharmacodynamics effect of nerve growth factor-loaded nanoparticles to induce PC12 cells

作　　者：包国庆[1] 龙大宏[2] 陈艳[3] 刘菲菲[1] 张君度[1] 

机构地区：[1]广州医学院人体解剖教研室,广东省广州市510182 [2]广州医学院继续教育学院,广东省广州市510182 [3]广州医学院第二附属医院康复科,广东省广州市510260

出　　处：《中国组织工程研究》2013年第16期2891-2898,共8页Chinese Journal of Tissue Engineering Research

基　　金：广东省自然科学基金(9151063101000016);广州市科技局应用基础研究计划项目(2009J1-C361-2);广州市属高校科研计划项目(2012C042)~~

摘　　要：背景:研究发现经表面修饰过的聚合物纳米粒能通过血脑屏障,可以改善药物对中枢神经系统疾病的疗效。目的:以生物可降解材料聚乙二醇-聚乳酸聚乙醇酸共聚物制备高包封率的载神经生长因子的纳米粒,并初步探讨其对PC12细胞的体外诱导效果评价。方法:采用复乳化溶剂扩散法制备载牛血清白蛋白的聚乙二醇-聚乳酸聚乙醇酸共聚物纳米粒,用单因素分析及正交设计对工艺进行优化筛选;扫描电镜观察纳米粒形态;纳米粒分析仪测定平均粒径和分散指数;BCA法测定纳米粒包封率及载药量,并进一步研究纳米粒体外释放特性。得到最好的制备方案后,制备载神经生长因子的聚合物纳米粒,并以此处理PC12细胞,倒置荧光显微镜观察载神经生长因子的聚乙二醇-聚乳酸聚乙醇酸共聚物纳米粒对PC12细胞的体外诱导状况,对其诱导效果、毒性及缓释效果进行评价。结果与结论:最优处方制备的载牛血清蛋白的聚乙二醇-聚乳酸聚乙醇酸共聚物纳米粒呈球形、大小均匀,平均粒径(258.9±5.73)nm,包封率为(80.56±2.23)%,内水相中投药量10mg时载药量约(4.24±0.12)%,体外释放符合Higuchi方程,分初期突释释放和后期缓释释放2个阶段,0-56d的累积释放总量分别为76.61%(牛血清白蛋白)、62.34%(神经生长因子)。载神经生长因子的聚乙二醇-聚乳酸聚乙醇酸共聚物纳米粒可以诱导PC12细胞像神经元样分化效能,表现出良好的缓释性能及无毒性作用。提示制备的载神经生长因子的聚乙二醇-聚乳酸聚乙醇酸共聚物纳米粒理化性质优良,在体外有良好的缓释效能及无毒性作用。BACKGROUND：It is found that the surface-modified polymer nanoparticles can pass through the blood-brain barrier, and improve the drug effects on the central nervous system diseases. OBJECTIVE：To prepare nerve growth factor-loaded high encapsulation efficiency nanoparticles with a iodegradable material, polyethylene glycol-polylactic acid glycolic acid copolymer, and to explore its effect on PC12 cel s. METHODS：Bovine serum albumin-loaded polyethylene glycol-polylactic acid glycolic acid copolymer nanoparticles were prepared using complex emulsion solvent diffusion method. The production process was screened by univariate analysis and orthogonal design method. The nanoparticle morphology was observed under scanning electron microscopy, and the average particle diameter and the dispersion index were measured by nanoparticle analyzer. The nanoparticle encapsulation efficiency and drug loading were analyzed by BCA method. Nanoparticle release in vitro was also investigated. After optimization of the preparation program, nerve growth factor-loaded polymer nanoparticles were prepared and applied to PC12 cel s. The cel s were then observed by inverted fluorescence microscope, and the induction effects, toxicity and slow-release effect of polyethylene glycol-polylactic acid glycolic acid copolymer nanoparticles loaded with nerve growth factor were evaluated. RESULTS AND CONCLUSION：Bovine serum albumin-loaded polyethylene glycol-polylactic acid glycolic acid copolymer nanoparticles were prepared by optimal method. The nanoparticles were spherical, and uniform in size, with an average particle size of （258.9±5.73） nm, and the encapsulation efficiency was （80.56±2.23）%. When the dosage within the aqueous phase was 10 mg, the drug loading was （4.24±0.12）%. The in vitro release of nanoparticles was in accordance with the Higuchi equation of. Release of the nanoparticles could be divided into two stages：initial burst release stage and late sustained release. The total cumulative release amount of bovin

关 键 词：生物材料 纳米生物材料 神经生长因子 聚乙二醇-聚乳酸聚乙醇酸共聚物 纳米药物 复乳化溶剂扩散法 纳米粒 PC12细胞 省级基金 

分 类 号：R318[医药卫生—生物医学工程]