Transfection efficiency of depolymerized chitosan and epidermal growth factor conjugated to chitosan-DNA polyplexes.
An efficient non-viral gene delivery for varieties of cells has been considered essential for gene therapy and tissue engineering. This study evaluated transfection efficiency of chitosan (HW) with molecular weights (Mw) at 470 and degree of deacetylation (DDA) 80% and its depolymerization product (LW) with Mw at 16 kDa and DDA 54%, as well as epidermal growth factor (EGF) conjugated to chitosan-DNA microparticles of both HW and LW by using either disulfide linkage or NHS-PEO(4)-Maleimide as a cross linker. The results revealed that the depolymerized LW at chitosan/DNA charge ratio 56:1 and pH 6.9 gave high transfection efficiency in both KB, a cancer cell line, and fibroblast cells at about the same level of Lipofectamine, but the EGF-conjugated chitosan-DNA polyplexes from these methods did not improve transfection efficiency, which may come from the aggregation and fusing of the complexes as shown in scanning electron microscopy. However, this depolymerized LW chitosan showed the potential for further development as a safe and cost-effective non-viral gene delivery vehicle.
Supaprutsakul
Sasamon
S
Center of Genomics and Bioinformatics Research, Faculty of Science, Prince of Songkla University, Hat-Yai, Songkhla, 90112, Thailand.
Chotigeat
Wilaiwan
W
Wanichpakorn
Supreya
S
Kedjarune-Leggat
Ureporn
U
D000941:Antigens / Q000378:metabolism; D002460:Cell Line; D048271:Chitosan / Q000737:chemistry* / Q000378:metabolism*; D004247:DNA / Q000008:administration & dosage / Q000737:chemistry / Q000378:metabolism; D004815:Epidermal Growth Factor / Q000378:metabolism; D018014:Gene Transfer Techniques; D005796:Genes; D015316:Genetic Therapy; D006801:Humans; D008565:Membrane Proteins; D008970:Molecular Weight; D009687:Nuclear Proteins; D014162:Transfection / Q000379:methods*
D000941:Antigens; D008565:Membrane Proteins; C488621:NHS protein, human; D009687:Nuclear Proteins; D004815:Epidermal Growth Factor; D004247:DNA; D048271:Chitosan