Chitosan-based vector/DNA complexes for gene delivery : Biophysical characteristics and transfection ability

Chitosan, a natural cationic polysaccharide, is a candidate non-viral vector for gene delivery. With the aim of developing this system, various biophysical characteristics of chitosan-condensed DNA complexes were measured, and transfections were performed. Transmission electronic microscopy (TEM) vi...

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Bibliographic Details
Published in:Pharmaceutical research 1998-09, Vol.15 (9), p.1332-1339
Main Authors: ERBACHER, P, SHAOMIN ZOU, BETTINGER, T, STEFFAN, A.-M, REMY, J.-S
Format: Article
Language:English
Subjects:
Biological and medical sciences
Biopolymers - genetics
Chitin - analogs & derivatives
Chitin - genetics
Chitin - metabolism
Chitin - ultrastructure
Chitosan
DNA - genetics
DNA - metabolism
DNA - ultrastructure
Gene Transfer Techniques
General pharmacology
Genetic Therapy
Genetic Vectors
HeLa Cells
Humans
Lectins - metabolism
Life Sciences
Medical sciences
Other
Pharmaceutical technology. Pharmaceutical industry
Pharmacology. Drug treatments
Polyethyleneimine - metabolism
Transfection
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Summary:Chitosan, a natural cationic polysaccharide, is a candidate non-viral vector for gene delivery. With the aim of developing this system, various biophysical characteristics of chitosan-condensed DNA complexes were measured, and transfections were performed. Transmission electronic microscopy (TEM) visualizations, sedimentation experiments, dynamic light scattering (DLS), and zeta potential measurements were realized. Transfections were made by using the luciferase reporter gene. In defined conditions, plasmid DNA formulated with chitosan produced homogenous populations of complexes which were stable and had a diameter of approximately 50-100 nm. Discrete particles of nicely condensed DNA had a donut, rod, or even pretzel shape. Chitosan/DNA complexes efficiently transfected HeLa cells, independently of the presence of 10% serum, and did not require an added endosomolytic agent. In addition, gene expression gradually increased over time. from 24 to 96 hours, whereas in the same conditions the efficacy of polyethylenimine-mediated transfection dropped by two orders of magnitude. At 96 hours, chitosan was found to be 10 times more efficient than PEI. However, chitosan-mediated transfection depended on the cell type. This dependency is discussed here. Chitosan presents some characteristics favorable for gene delivery, such as the ability to condense DNA and form small discrete particles in defined conditions.
ISSN:0724-8741
1573-904X
DOI:10.1023/a:1011981000671