Poly(ethylene imine)-g-chitosan using EX-810 as a spacer for nonviral gene delivery vectors.
Polyelectrolyte complexes have been widely studied as gene carriers in recent years. In this study, poly (ethylene imine) was grafted onto chitosan (PEI-g-CHI) as a nonviral gene carrier in order to improve the water solubility as well as the inherent transfection efficiency of chitosan. We present a novel method to conjugate the amine or hydroxyl groups of chitosan (CHI) and the amine groups of PEI through opening the epoxide rings of ethylene glycol diglycidyl ether (EX-810), which also brings the merits as mentioned in PEGylation chemistry. The degree of substitution of PEI onto CHI was characterized by NMR. The preliminarily cellular mechanisms, from the cellular entry to the endosomal release, were investigated by the correlations among the physicochemical properties of the DNA-polymer complexes, the buffering capacity of the modified polymer, the cytotoxicity, and the efficiency of the transgene expression. The cytotoxicity assayed by MTT shows that cell viability of PEI-g-CHI is higher than CHI especially noticeable at high concentrations using human kidney 293T cells. The efficiency of transgene expression and the amount of intracellular plasmid were monitored using green fluorescent protein (GFP) and visualized by fluorescence microscopy. The transfection efficiency of PEI-g-CHI/DNA polyplex is significantly better than CHI/DNA polyplex when using the weight ratios higher than 2.5.
Lou
Yu-Lun
YL
Faculty of Medicinal and Applied Chemistry, School of Life Science, Kaohsiung Medical University, Kaohsiung, Taiwan 80708, Republic of China.
Peng
Yu-Shiang
YS
Chen
Bing-Hung
BH
Wang
Li-Fang
LF
Leong
Kam W
KW
D001672:Biocompatible Materials / Q000737:chemistry*; D002460:Cell Line; D002470:Cell Survival; D048271:Chitosan / Q000737:chemistry*; D004247:DNA / Q000737:chemistry / Q000378:metabolism; D004853:Epoxy Resins / Q000737:chemistry*; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000737:chemistry*; D006801:Humans; D007097:Imines / Q000737:chemistry*; D008422:Materials Testing; D015394:Molecular Structure; D010316:Particle Size; D011095:Polyethylenes / Q000737:chemistry*; D019076:Transgenes
D001672:Biocompatible Materials; D004853:Epoxy Resins; D007097:Imines; D011095:Polyethylenes; C505405:poly(ethylene imine); C035364:Quetol 651; D004247:DNA; D048271:Chitosan