摘要
首次研究了不同均匀度磁场对磁转染效果的影响。测定了单边Halbach磁体和商品化96孔磁板的磁场均匀度。以化学共沉淀法制备聚乙烯亚胺(Polyethyleneimine,PEI)修饰的四氧化三铁(Fe_3O_4)纳米颗粒(PEI-Fe_3O_4),并用透射电子显微镜(TEM)、振动样品磁强计(VSM)、原子力显微镜、琼脂糖凝胶电泳等对其形貌、组成、DNA结合能力等进行表征。用倒置荧光显微镜、流式细胞术观察不同均匀度磁场下人肾上皮细胞(HEK293)对带有绿色荧光蛋白报告基因(GFP)的质粒pDNA(pAdTrackOK)的表达效果,并采用TEM观察PEI-Fe_3O_4磁性纳米颗粒进入细胞的过程。结果显示,所选取的两种磁场均匀度相差约100倍。制备的PEI-Fe_3O_4纳米复合物具有超顺磁性,对质粒pDNA(pAdTrack-OK)有较好的复合能力,其最佳结合氮磷比为0.5;流式细胞术显示转染效率为PEI-Fe_3O_4-pDNA+均匀磁场组(77.75%±0.07%)>PEI-Fe_3O_4-pDNA+不均匀磁场组(30.65%±0.49%)>PEI-Fe_3O_4-pDNA不加磁场组(7.90%±0.56%)(p<0.05);PEI-Fe_3O_4磁性纳米颗粒能有效被细胞吞噬,且对细胞形态的影响不大。结果表明,当磁场强度一定时,磁场均匀度越高,磁转染效率越高,单边Halbach磁体与磁转染结合可以作为一种提高转染效率的新手段,也可以进一步应用在基因治疗中。
The effect of uniform magnetic field(MF)on magnetofection was studied for the first time.The uniformity of the unilateral Halbach magnet and the commercialized 96 hole magnet were measured.Polyethyleneimine(PEI)modified ferroferric oxide(Fe 3O 4)nanoparticles(PEI-Fe 3O 4)were prepared by chemical coprecipitation.The morphology,composition and the DNA binding ability of PEI-Fe 3O 4 were charaterized by transmission electron microscope(TEM),vibrating sample magnetometer(VSM),atomic force microscopy and agarose gel electrophoresis.The green fluorescent protein(GFP)expression efficiency in human renal epithelial cells(HEK293)under different uniformity of magnetic field was detected with inverted fluorescence microscope and flow cytometry.The phagocytosis of PEI-Fe 3O 4 nanoparticles was observed by TEM.The results showed that the difference of uniformity of the two magnetic fields was about 100 times.The superparamagnetic PEI-Fe 3O 4 has a good compound ability for plasmid pDNA,and its optimal combination ratio is N/P=0.5.Flow cytometry showed that the transfection efficiency was:PEI-Fe 3O 4-pDNA+uniform MF group(77.75%±0.07%)>PEI-Fe 3O 4-pDNA+nonuniform MF group(30.65%±0.49%)>PEI-Fe 3O 4-pDNA without MF group(7.90%±0.56%)(p<0.05).PEI-Fe 3O 4 magnetic nanoparticles can be effectively consumed by cells and have little effect on cell morphology.It can be concluded that when the magnetic field intensity is certain,higher uniformity of magnetic field contributes to the higher the magnetic transfection efficiency.The combination of the unilateral Halbach magnet and the magnetosfection can be used as a new method to improve the transfection rate,which may potentially be employed in gene therapy.
作者
谢丽娜
罗聪
吴嘉敏
王昌绚
邬均
XIE Lina;LUO Cong;WU Jiamin;WANG Changxuan;WU Jun(Ministry of Education Key Laboratory of Child Development and Disorders,Pediatric Research Institute,Children’s Hospital of Chongqing Medical University,Chongqing 400014;Department of Orthopaedics,Children’s Hospital of Chongqing Medical University,Chongqing 400014;China International Science and Technology Cooperation Base of Child Development and Critical Disorders,Pediatric Research Institute,Children’s Hospital of Chongqing Medical University,Chongqing 400014;Key Laboratory of Pediatrics,Pediatric Research Institute,Children’s Hospital of Chongqing Medical University,Chongqing 400014;State Key Laboratory of Power Transmission Equipment and System Security and New Technology,School of Electrical Engineering,Chongqing University,Chongqing 400044)
出处
《材料导报》
EI
CAS
CSCD
北大核心
2018年第8期1247-1251,共5页
Materials Reports
基金
国家自然科学基金(30973062)
国家临床重点专科建设项目(国卫办医函[2013]544)
关键词
磁转染
均匀磁场
不均匀磁场
超顺磁性纳米颗粒
magnetofection
uniform magnetic field
nonuniform magnetic field
magnetic nanoparticles
