Establishing Gene Delivery Systems Based on Small-Sized Chitosan Nanoparticles

摘要

Chitosan is a natural cationic polysaccharide,which is often used for preparing biomedical materials because of its high biocompatibility.In this study,chitosan with a molecular weight of 160 k Da was chosen to prepare chitosan nanoparticles(CSNPs) as gene vectors by ionic cross-linking with tripolyphosphate(TPP).CSNPs were characterized in terms of particle size,zeta potential,and polydispersity index(PDI) using a Zetasizer,and morphology was evaluated by transmission electron microscopy(TEM).Furthermore,the cytotoxicity and biocompatibility of CSNPs were correspondingly examined by a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT) assay and histological examination.Agarose gel electrophoresis and UV spectrophotometric methods were performed to measure the loading capacity.The cell transfection efficiency of CSNPs loaded with plasmids or si RNA was analyzed by fluorescence microscopy or laser scanning confocal microscopy.The results showed that CSNPs were prepared successfully by the ionic gelation method,which had a smaller particle size(100 nm-200 nm),stable dispersibility,low cytotoxicity,good tissue-biocompatibility,and high gene-loading efficiency.These CSNPs could transfer the plasmids or si RNA to cells.However,CSNPs might have a much higher transfection efficiency for si RNAs than for plasmids,which implies that CSNPs might be a safer and more efficient vector for delivering si RNAs rather than plasmids.