多孔壳聚糖/明胶复合材料载入治疗基因成分的实验  

Experiment of loading the plasmid DNA into porous chitosan/gelatin matrices

作　　者：李黎[1] 曹玲艳[1] 张秋红[1] 张俊峰[2] 

机构地区：[1]解放军南京军区南京总医院制剂科,江苏省南京市210002 [2]南京大学生物医药国家重点实验室,江苏省南京市210093

出　　处：《中国组织工程研究与临床康复》2007年第5期807-810,共4页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金：江苏省自然科学基金面上项目资助项目(BK2005085)~~

摘　　要：目的:探讨多孔壳聚糖/明胶复合生物材料载入治疗基因的可行性。方法:实验于2003-09/2005-04在南京大学国家重点生物医药技术实验室完成。实验材料:壳聚糖(Mr约160000,脱乙酰化程度85%,Sigma公司),明胶B(取自牛皮肤,Sigma公司),能够在真核体系表达的质粒DNA(编码有转化生长因子β1,pCMV-TGF-β1)。1mg支架材料DNA控制在2μg,通过二次冻干的方式制备含有pCMV-TGF-β1的多孔壳聚糖/明胶三维架材料,观测多孔支架材料携带基因成分的微观形态、体外释放特性以及材料对质粒DNA的保护作用。结果:壳聚糖与明胶材料具有良好的成孔特性,孔隙较均一,材料表面光整;材料在浸泡磷酸盐缓冲液后,1～3d内快速释放,DNA迅速达到释放浓度峰值,并能够维持3周的释放时间;材料对基因成分有保护作用。结论:多孔壳聚糖/明胶三维支架材料能够携带质粒DNA。通过二次快速冻干方式制备基因活化材料可以保护基因成分不被破坏,并可保持一定浓度的持续释放时间,为基因活化材料进一步在体内运用提供了依据。AIM： To explore the feasibility and methods of plasmid DNA loaded into porous chitosan/gelation matrices. METHODS： The experiment was Conducted in the State Key Laboratory of Pharmaceutical Biotechnology, Nanjing University between September 2003 and April 2005. The experiment materials： chitosan （Mr 160 000, deacetylation 85%, Sigma Company）, gelation B （came from buffalo-hide, Sigma Company）, plasmid DNA could express in eukarya system （transforming growth factor β 1, pCMV-TGF-β 1）. 1 mg scaffold material DNA was limited in 2 μg. The porous chitosan/gelation matrices three-dimensional scaffolds containing pCMV-TGF-β 1 was prepared after twice freeze-dry and the micromorphous of the gene in the material, release and the protection of the material to plasmid DNA ware observed. RESULTS： Porous chitosan/gelation matrices had good wall-formation feature, and the wall was even, the surface was smooth; after soaked in buffer phosphate solution, the material released in 1-3 days, and DNA rapidly reached the concentration peak, which could be lasted for 3 weeks. The material had protective effect on gene components. CONCLUSION： The porous chitosan/gelation matrices three-dimensional scaffolds load with plasmid DNA. The gene activated material after twice freeze-dry can prevent the gene components from damage, and sustain release time of plasmid DNA, which provides an evidence for the further application of gene-activated material in vivo.

关 键 词：壳聚糖 明胶 质粒 DNA 

分 类 号：R318.08[医药卫生—生物医学工程]