Water-soluble and low molecular weight chitosan-based plasmid DNA delivery

Chitosan, a natural cationic polysaccharide, is a candidate non-viral vector for gene delivery because of its high positive charges and low cytotoxicity. In this study, low molecular weight chitosan (LMWC, molecular weight of 22 kDa) was characterized and evaluated as a gene carrier. Plasmid/LMWC co...

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Bibliographic Details
Published in:Pharmaceutical research 2001-04, Vol.18 (4), p.427-431
Main Authors: MINHYUNG LEE, NAH, Jae-Woon, YOUNGMIN KWON, JAE JOON KOH, KYUNG SOO KO, SUNG WAN KIM
Format: Article
Language:English
Subjects:
Antineoplastic Agents - administration & dosage
Biological and medical sciences
Cell Line
Chitin - administration & dosage
Chitin - analogs & derivatives
Chitin - genetics
Chitosan
Cytotoxicity
DNA - administration & dosage
DNA - genetics
Drug Delivery Systems - methods
Efficiency
Gene therapy
General pharmacology
Humans
Medical sciences
Molecular weight
Pharmaceutical technology. Pharmaceutical industry
Pharmacology. Drug treatments
Physiology
Plasmids
Plasmids - administration & dosage
Plasmids - genetics
Polymers
Toxicity
Transfection - methods
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Summary:Chitosan, a natural cationic polysaccharide, is a candidate non-viral vector for gene delivery because of its high positive charges and low cytotoxicity. In this study, low molecular weight chitosan (LMWC, molecular weight of 22 kDa) was characterized and evaluated as a gene carrier. Plasmid/LMWC complex was analyzed in 1% agarose gel electrophoresis. To confirm that the LMWC protected plasmids from nuclease. DNase I protection assays were performed. pSV-beta-galactosidase plasmid/LMWC complex was transfected into 293T cells and transfection efficiency was evaluated by beta-galactosidase assay. Cytotoxicity of LMWC was determined by MTT assay. Unlike high molecular weight chitosan (HMWC), LMWC is highly water soluble, and can form complex with plasmids in physiological buffer. The plasmid DNA was completely retarded at a weight ratio of 1:2 (plasmid:LMWC) in 1% agarose gel. DNase I protection assay showed that plasmids were protected from DNase-I over 60 min. The most efficient transfection was obtained at a weight ratio of 1:3 (plasmid:LMWC). The transfection efficiency of LMWC was significantly higher than naked DNA and higher than poly-L-lysine (PLL). MTT assay showed that LMWC was less cytotoxic than PLL. LMWC is non-toxic and has higher transfection efficiency than PLL. Therefore, LMWC will be useful in the development of safe gene carriers.
ISSN:0724-8741
1573-904X
DOI:10.1023/A:1011037807261