Tailoring of chitosans for gene delivery: novel self-branched glycosylated chitosan oligomers with improved functional properties.
Chitosan is a promising biomaterial with an attractive safety profile; however, its application potential for gene delivery is hampered by poor compatibility at physiological pH values. Here we have tailored the molecular architecture of chitosan to improve the functional properties and gene transfer efficacy of chitosan oligomers and have developed self-branched glycosylated chitosan oligomer (SB-TCO) substituted with a trisaccharide containing N-acetylglucosamine, AAM. SB-TCO was prepared by controlled depolymerization of chitosan, followed by simultaneous branching and AAM substitution. The product was fully soluble at physiological pH and complexed plasmid DNA into polyplexes of high colloidal and physical stability. SB-TCO displayed high transfection efficacy in HEK293 cells, reaching transfection efficiencies of up to 70%, and large amounts of transgene were produced. Gene transfer efficacy was confirmed in HepG2 cells, where gene expression levels mediated by SB-TCO were up to 10 and 4 times higher than those obtained with unsubstituted and substituted linear oligomers, respectively. The rapid onset of transgene expression in both cell lines indicates efficient DNA release and transcription from SB-TCO polyplexes. In comparison with 22 kDa linear PEI-based transfection reagent used as the control, SB-TCO possessed higher gene transfer efficacy, significantly lower cytotoxicity, and improved serum compatibility.
Norwegian Biopolymer Laboratory, Department of Biotechnology, Norwegian University of Science and Technology, 7491 Trondheim, Norway. sabina.strand@biotech.ntnu.no
Sabina P
SP
Strand
Mohamed M
MM
Issa
Bjørn E
BE
Christensen
Kjell M
KM
Vårum
Per
P
Artursson
D001672:Biocompatible Materials / Q000737:chemistry; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D048271:Chitosan / Q000737:chemistry*; D018014:Gene Transfer Techniques; D006031:Glycosylation; D006801:Humans; D010957:Plasmids; D012995:Solubility
D001672:Biocompatible Materials; D048271:Chitosan