CSIR-Institute of Genomics and Integrative Biology, Delhi University, Delhi, India.
Nanomedicine. 2012 Apr;8(3):337-45. doi: 10.1016/j.nano.2011.06.022. Epub 2011 Jul 12.
Chitosan was partially converted to its chlorohydrin derivative by the reaction with epichlohydrin, which was subsequently reacted with varying amounts of lPEI(2.5 kD) to obtain a series of chitosan-lPEI(2.5 kD) copolymers (CP). These copolymers were then characterized and evaluated in terms of transfection efficiency (in vitro and in vivo), cell viability, DNA release and buffering capacity. The CP-4 copolymer (the best among the CP series) showed enhanced transfection (-2 - 24 folds) in comparison with chitosan, lPEI(2.5 kD), bPEI(25 kD) and Lipofectamine in HEK293, HeLa and CHO cells. The buffering capacity (in the pH range of 3 - 7.5), as shown by confocal microscopy, and DNA-release capability of the CP copolymers, was found to be significantly enhanced over chitosan. Intravenous administration of CP-4/DNA polyplex in mice followed by the reporter gene analysis showed the highest gene expression in spleen. Collectively, these results demonstrate the potential of CP-4 copolymer as a safe and efficient nonviral vector. From the Clinical Editor: Chitosan -PEI (2.5 kD) copolymers (CP) were characterized and their transfection efficiency, DNA release and buffering capacity were studied. The CP-4 copolymer significantly enhanced buffering capacity and provided the highest gene expression levels. The method may be used to enhance DNA transfection.
壳聚糖与表氯醇反应部分转化为其氯醇衍生物，然后与不同量的 lPEI（2.5kDa）反应，得到一系列壳聚糖-lPEI（2.5kDa）共聚物（CP）。这些共聚物随后进行了表征，并从转染效率（体外和体内）、细胞活力、DNA 释放和缓冲能力等方面进行了评估。与壳聚糖、lPEI（2.5kDa）、bPEI（25kDa）和 Lipofectamine 相比，CP-4 共聚物（CP 系列中表现最好的）在 HEK293、HeLa 和 CHO 细胞中表现出增强的转染效率（-2 至 24 倍）。共聚物 CP 的缓冲能力（在 pH 值 3-7.5 范围内）和 DNA 释放能力通过共聚焦显微镜显示出明显增强。CP-4/DNA 聚合物在小鼠体内静脉给药后，通过报告基因分析显示脾脏中的基因表达最高。总的来说，这些结果表明 CP-4 共聚物作为一种安全有效的非病毒载体具有潜力。来自临床编辑：壳聚糖-PEI（2.5kDa）共聚物（CP）进行了表征，研究了它们的转染效率、DNA 释放和缓冲能力。CP-4 共聚物显著增强了缓冲能力，并提供了最高的基因表达水平。该方法可用于增强 DNA 转染。
