中国组织工程研究

中国组织工程研究 ›› 2012, Vol. 16 ›› Issue (21): 3929-3932.doi: 10.3969/j.issn.1673-8225.2012.21.030

• 组织构建基础实验 basic experiments in tissue construction • 上一篇    下一篇

水/油/水双乳化技术制备高效性神经生长因子***☆

谷海刚1，李晓滨1，龙大宏2   

  1. 广州医学院，1组织学与胚胎学教研室，2解剖学教研室，广东省广州市   510182
  • 出版日期:2012-05-20 发布日期:2012-05-20
  • 作者简介:谷海刚☆，男，1976年生，河南省洛阳市人，汉族，2007年北京协和医学院-清华大学医学部毕业，博士，讲师，主要从事神经系统疾病治疗干细胞、非病毒基因载体和新型给药系统的研究。guhaigang@yahoo.com.cn
  • 基金资助:

    国家自然科学基金项目(30800345)，课题名称：载神经生长因子基因纳米粒子对老年性痴呆的治疗作用及其机理。广东省医学科研基金项目(B2008071)，课题名称：PEI/DNA-NGF纳米粒子治疗老年性痴呆的实验研究。广州医学院基金项目(0706059)，课题名称：复合神经营养素缓释微球治疗老年性痴呆的实验研究。

High efficient nerve growth factor made by double emulsion water/oil/water method

Gu Hai-gang1, Li Xiao-bin1, Long Da-hong2   

  1. 1Department of Human Histology and Embryology, 2Department of Human Anatomy, Guangzhou Medical College, Guangzhou  510182, Guangdong Province, China
  • Online:2012-05-20 Published:2012-05-20
  • About author:Gu Hai-gang☆, Doctor, Lecturer, Department of Histology and Embryology, Guangzhou Medical College, Guangzhou 510182, Guangdong Province, China guhaigang@ yahoo.com.cn
  • Supported by:

     the National Natural Science Foundation of China, No.30800345*; Medical Scientific Research Foundation of Guangdong Province, No.B2008071*; Foundation from Guangzhou Medical College, No.0706059*

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摘要：

背景：神经生长因子是大分子蛋白类物质，很难透过血脑屏障，基因治疗可以有效地将神经营养因子投递到脑内。
目的：探索制备高效性神经生长因子基因纳米粒子的方法。
方法：利用水/油/水(W/O/W)双乳化技术制备神经生长因子基因纳米粒子，用壳聚糖对其进行表面修饰，同时偶联Tat多肽。切断穹隆-海马伞诱使基底前脑胆碱能神经元溃变。16只SD雄性大鼠随机数字表均分成损伤组和基底前脑显微注射神经生长因子基因的纳米粒子组。
结果与结论：制备的纳米粒子粒径分布为110~150 nm。壳聚糖可明显提高其表面正电荷和包封率，但对其粒径大小影响不大。偶联Tat多肽则明显提高其转染效率。说明利用水/油/水技术可以成功制备神经生长因子基因纳米粒子。该粒子移植后对基底前脑有明显的保护作用。
 

关键词: 神经生长因子基因纳米粒子, 聚乳酸-聚乳二醇酸, 壳聚糖, Tat多肽, 转染效率

Abstract:

BACKGROUND: Nerve growth factor (NGF) is a macromolecular protein that cannot easily cross the blood-brain barrier. Gene therapy can successfully deliver neurotrophic factors into the brain.
OBJECTIVE: To explore how to formulate the high efficient NGF gene nanoparticles.
METHODS: NGF nanoparticles were prepared by double emulsion water/oil/water method. Chitosan and Tat polypeptide were used to modify NGF nanoparticles. Neuron degeneration of cholinergic neurons in the rat’s basal forebrain was induced by fimbria-fornix transection. Sixteen SD rats were randomly divided into injury group and nanoparticles group micro injected with NGF gene in the basal forebrain. 
RESULTS AND CONCLUSION: The distribution of particle size of NGF nanoparticles was from 110 nm to 150 nm. Chitosan could increase zeta potential and loading efficiency of NGF nanoparticles, but not particle size. Tat polypeptide could significantly increase the transfection efficiency of NGF nanoparticles. It indicated that water/oil/water method could successfully make the NGF nanoparticles. And the NGF nanoparticles transplantation could protect the basal forebrain.

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