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.
Highlights
IntroductionGene therapy has attracted substantial attention due to its vast therapeutic potential for the treatment of various diseases, including genetic disorders, cancer, and infections [1]
Gene therapy has attracted substantial attention due to its vast therapeutic potential for the treatment of various diseases, including genetic disorders, cancer, and infections [1].Another application of gene therapy is the use of DNA as a vaccine to induce immunity against infectious diseases and cancers
Previous studies unambiguously demonstrated a strong dependence of the key characteristics
Summary
Gene therapy has attracted substantial attention due to its vast therapeutic potential for the treatment of various diseases, including genetic disorders, cancer, and infections [1]. Another application of gene therapy is the use of DNA as a vaccine to induce immunity against infectious diseases and cancers. Gene delivery systems can be broadly divided into two categories: recombinant virus vectors and synthetic vectors. Viral vectors have shown good transduction efficiency in in vivo experiments, but their use in clinical practice is severely limited because of their serious side effects (e.g., immunogenicity, carcinogenicity, etc.) and the difficulties associated with vector modification [6]
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