PDF(Pubmed)
Abstract：
In this paper, we compared the transfection efficiency and cytotoxicity of methylglycol-chitosan (MG-CS) and diethylaminoethyl-chitosan (DEAE-CSI and DEAE-CSII with degrees of substitution of 1.2 and 0.57, respectively) to that of Lipofectamine (used as a reference transfection vector). MG-CS contains quaternary amines to improve DNA binding, whereas the DEAE-CS exhibits pH buffering capability that would ostensibly enhance transfection efficiency by promoting endosomal escape. Gel retardation assays showed that both DEAE-CS and MG-CS bound to DNA at a polysaccharide:DNA mass ratio of 2:1. In Calu-3 cells, the DNA transfection activity was significantly better with MG-CS than with DEAE-CS, and the efficiency improved with increasing polysaccharide:DNA ratios. By contrast, the efficiency of DEAE-CSI and DEAE-CSII was independent of the polysaccharide:DNA ratio. Conversely, in the transfection-recalcitrant JAWSII cells, both Lipofectamine and MG-CS showed significantly lower DNA transfection activity than in Calu-3 cells, whereas the efficiency of DEAE-CSI and DEAE-CSII was similar in both cell lines. The toxicity of DEAE-CS increased with increasing concentrations of the polymer and its degree of substitution, whereas MG-CS demonstrated negligible cytotoxicity, even at the highest concentration studied. Overall, MG-CS proved to be a more efficient and less toxic transfection agent when compared to DEAE-CS.
摘要：
在本文中,我们比较了聚乙二醇-壳聚糖(MG-CS)和二乙基氨基乙基-壳聚糖(DEAE-CSI和DEAE-CSII,取代度分别为1.2和0.57)与Lipofectamine(用作参考转染载体)的转染效率和细胞毒性.MG-CS含有季胺以改善DNA结合，而DEAE-CS表现出pH缓冲能力，表面上通过促进内体逃逸来提高转染效率。凝胶阻滞分析显示,DEAE-CS和MG-CS均以2:1的多糖:DNA质量比与DNA结合。在Calu-3细胞中,MG-CS的DNA转染活性明显优于DEAE-CS，并且效率随着多糖:DNA比率的增加而提高。相比之下,DEAE-CSI和DEAE-CSII的效率与多糖：DNA比无关。相反，在转染顽固的JAWSII细胞中，Lipofectamine和MG-CS均显示出显著低于Calu-3细胞的DNA转染活性,而DEAE-CSI和DEAE-CSII在两种细胞系中的效率相似。DEAE-CS的毒性随着聚合物浓度及其取代度的增加而增加，而MG-CS表现出可忽略的细胞毒性，即使在研究的最高浓度。总的来说,与DEAE-CS相比，MG-CS被证明是更有效且毒性更低的转染剂。
