临床神经外科杂志Issue(2)：109-114,6.DOI:10.3969/j.issn.1672-7770.2019.02.004
载 BDNF 基因 PEG-PLGA 纳米微球的制备及其转染外源性神经干细胞的研究
Preparation of PEG-PLGA-BDNF nanoparticles and exploratory study on transfection of
摘要
Abstract
Objective To construct nano-gene microspheres(PEG-PLGA-BDNF-NPs) with the function of express nerve repair protein , and explore the effects of transfection on exogenous neural stem cells ( NSCs) on their biological functions.Methods PEG-PLGA nanoparticles transfected with BDNF gene plasmids were prepared by double emulsion-solvent evaporation method.The morphology of nanoparticles was observed by scanning electron microscopy and transmission electron microscope.The particle size and Zeta potential of nanoparticles were measured by laser particle size analyzer.The entrapment efficiency of PEG-PLGA nanoparticles and the drug loading of BDNF gene plasmids were examined.The in vitro release plasmid of PEG-PLGA nanoparticle transfection complex was tested.The biocompatibility of the nanoparticle transfection complex and the effect of the nanoparticle complex on the morphology of NSCs were detected by CCK-8.The secretion of neural stem cells after transfection of nanoparticles transfected complex was observed by inverted fluorescence microscope.Western blot and quantitative RT-PCR were used to detect the secretion of BDNF.Results (1) The gene-loaded PEG-PLGA nanoparticles were prepared.The encapsulation efficiency and drug loading were high , which had a certain sustained release effect.(2) The neural stem cells with the ability to proliferate and express the nestin protein are isolated and cultured , and can be induced to differentiate into neurons and glial cells.(3) PEG-PLGA nanoparticles containing BDNF gene were highly transfected into neural stem cells , and transfection did not cause significant cytotoxicity.Western-blot and RT-PCR showed that BDNF was abundantly expressed in these stem cells.Conclusions The rat neural stem cells were successfully extracted and cultured , and the PEG-PLGA-BDNF-NPs were prepared.An exogenous neural stem cell line loaded with PEG-PLGA-BDNF-NPs was first constructed to express BDNF protein with neurorestorative function .关键词
纳米微球/神经干细胞/基因治疗/转染Key words
nanoparticle/ neural stem cells/ gene therapy/ transfection分类
医药卫生引用本文复制引用
YU Yong-bo,TANG Jian,ZHANG Gui-long..载 BDNF 基因 PEG-PLGA 纳米微球的制备及其转染外源性神经干细胞的研究[J].临床神经外科杂志,2019,(2):109-114,6.基金项目
国家自然科学基金(81671819,81471780) （81671819,81471780）
江苏省六大人才高峰项目(2015-WSW-001) （2015-WSW-001）
