Hydrogel pullulan nanoparticles encapsulating pBUDLacZ plasmid as an efficient gene delivery carrier
Hydrogel pullulan nanoparticles encapsulating pBUDLacZ plasmid as an efficient gene delivery carrier
复制标题
DOI：
10.1016/j.jconrel.2004.06.016
复制
发表时间：
2004-09-14
影响因子：
10.8
通讯作者：
Gupta, AK
中科院分区：
文献类型：
--
作者：
Gupta, M;Gupta, AK
本研究提供了一种通过将核酸分子包封在水凝胶普鲁兰多糖纳米颗粒内来增强核酸分子向细胞的递送的方法。在这项研究中，普鲁兰多糖纳米粒子封装pBUDLacZ质粒已被制备内的水液滴的w/o微乳液。透射电子显微镜（TEM）图像显示颗粒为球形，尺寸为45 +/-0.80 nm直径。通过3-（4，5-二甲基噻唑-2-基）-2，5-二苯基-溴化四唑（MTT）测定法测定的细胞毒性研究表明，在高达1000 μ g/ml的纳米颗粒浓度下，与纳米颗粒孵育的细胞保持超过100%的存活率。从扫描电子显微镜图像中，观察到纳米颗粒被内化，并且由于纳米颗粒内化，细胞在细胞体中表现出空泡。纳米颗粒的内吞作用导致人成纤维细胞的F-肌动蛋白和β-微管蛋白细胞骨架的破坏。体外转染HEK293和COS-7细胞的功效表现出细胞类型依赖性，其中COS细胞具有更高的基因表达。普鲁兰多糖纳米颗粒在COS-7细胞中的β-gal表达与市售Lipofectamine 2000相当。本研究结果对于普鲁兰多糖纳米粒作为药物和基因的细胞内递送系统的发展是令人鼓舞的。(C)2004 Elsevier B.V.保留所有权利。
This study provides a method for enhancing the delivery of nucleic acid molecules to cells by encapsulating it inside the hydrogel pullulan nanoparticles. In this study, pullulan nanoparticles encapsulating pBUDLacZ plasmid have been prepared inside the aqueous droplets of w/o microemulsions. Transmission electron microscopy (TEM) image showed that the particles are spherical in shape with size of 45+/-0.80 nm diameter. Cell cytotoxicity studies as determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) assay demonstrated that cells incubated with nanoparticles remained more than 100% viable at nanoparticle concentration as high as 1000 mug/ml. From scanning electron microscope images, it was observed that the nanoparticles were internalised and the cells exhibited vacuoles in the cell body due to nanoparticle internalisation. Endocytosis of nanoparticles resulted in disruption of F-actin and beta-tubulin cytoskeleton of human fibroblasts. The efficacy of transfection in vitro on HEK293 and COS-7 cells demonstrated cell type dependence, with COS cells having a higher gene expression. The beta-gal expression in COS-7 cells by pullulan nanoparticle was comparable to commercially available Lipofectamine 2000. The results of this study are encouraging for the development of pullulan nanoparticles as an intracellular delivery system for drugs and genes. (C) 2004 Elsevier B.V. All rights reserved.