摘要
目的优化试验条件,制备合适的壳聚糖纳米粒,并连接上质粒,研究壳聚糖纳米粒对DNA的结合力。方法以离子交联法制备壳聚糖纳米粒,并送激光微粒度仪及扫描电子显微镜检测,了解粒径的分布与态;通过静电吸附作用连接上增强型绿色荧光蛋白表达质粒pEGFP-C1 (报告基因);经琼脂糖凝胶电泳分析体与DNA结合能力,并通过紫外分光光度计检测其载药量和包埋率。结果微粒度分析仪及电镜检测证实聚糖纳米粒呈均匀分散的球形颗粒,最小粒径为50 nm,平均直径为95 nm。琼脂糖凝胶电泳的结果显示壳聚纳米粒能有效地结合增强型绿色荧光蛋白表达质粒pEGFP-C1,紫外分光光度计检测不同比例结合(纳米:质粒)pEGFP-C1质粒的壳聚糖纳米粒的包埋率分别为:100%(50:10),100%(50:20),92%(50:75)和5%(50:100)。结论制备出粒径较小、均匀的壳聚糖纳米粒,并且壳聚糖纳米粒能有效地连接上质粒。
Objective: To prepare appropriate chitosan nanoparticles optimizing the experimental condition. To load it with plasmid and study its loading capability and protection ability to plasmid. Methods: To prepare the chitosan nanoparticles with cocervation process,to measure its diameter,the diametric distribution range and its form by scanning electron microscope and laser atomic examining apparatus. The enhanced green fluorescent protein as reporter gene is absorbed to chitosan nanoparticles by electrostaticforces. Results: Chitosan nanoparticles is showed to be spherical particles distributed evenly, the least nanoparticles is 50 nm in diameter, d(0.5) is 95nm,by scanning electron microscope and laser atomic examining apparatus; Agarose gel electrophoresis showed that the enhanced green fluorescent protein was absorbed to chitosan nanoparticles effectively. The encapsulation efficiency of nanoparticles-DNAmixed by different ratio(nanoparticles:plasmid) is 100%(50:10), 100%(50:20), 92%(50:75), 65%(50:100) by examining of fluorescent miscroscope. Conclusion: Homogeneous and small in diameter chitosan nanoparticles has been prepared and loaded with plasmid effectively.
出处
《中国现代医学杂志》
CAS
CSCD
2004年第19期48-51,共4页
China Journal of Modern Medicine
基金
国家"十五"863计划课题:"磁性纳米药物载体系统治疗肝脏恶性肿瘤研究"项目编号:2002AA214011
