Comparative Study of Diethylaminoethyl-Chitosan and Methylglycol-Chitosan as Potential Non-Viral Vectors for Gene Therapy

Abstract

Abstract:In this paper, we compared the transfection efficiency and cytotoxicity of methylglycolchitosan (MG-CS) and diethylaminoethyl-chitosan (DEAE-CS I and DEAE-CS II 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-CS I and DEAE-CS II 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-CS I and DEAE-CS II 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.

Evaluation of the mucoadhesive properties of chitosan nanoparticles prepared using different chitosan to tripolyphosphate (CS:TPP) ratios

“…Furthermore, the pH dependent solubility of chitosan enables tuneable pH-dependent release [9], which can be further tailored by derivatisation [10] or cross-linking (with TPP for example)…”

Synthesis, Bioapplications, and Toxicity Evaluation of Chitosan-Based Nanoparticles

“…The transfection efficiency of Ch-based nano-vehicles is cell-type-dependent; there is no toxicity with respect to other more toxic particles like lipofectamine, a cationic lipid; unfortunately, the transfection efficiency of Ch nanoparticles is lower than that of lipofectamine [187]. For an effective transfection, not only is the internalization important, but so is the subsequent endo-lysosomal escape [188].…”

Chitosan-Based Drug Delivery System: Applications in Fish Biotechnology

“…To increase the stability of chitosan-based formulations, a number of chitosan derivatives have been developed. Among them, PEGylation [35][36][37], glycosylation [3,38,39] and quaternization [39][40][41][42]. The choice of the method for preparing chitosan-nucleic acid complexes can also significantly affect stability of the complex and transfection efficiency.…”