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Relationship between the physical shape and the efficiency of oligomeric chitosan as a gene delivery system in vitro and in vivo
Author(s) -
KöpingHöggård M.,
Mel'nikova Y. S.,
Vårum K. M.,
Lindman B.,
Artursson P.
Publication year - 2003
Publication title -
the journal of gene medicine
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.689
H-Index - 91
eISSN - 1521-2254
pISSN - 1099-498X
DOI - 10.1002/jgm.327
Subject(s) - chitosan , gene delivery , in vivo , oligomer , chemistry , in vitro , biophysics , ionic strength , molecular mass , dna , reporter gene , transfection , gene expression , gene , biochemistry , polymer chemistry , organic chemistry , biology , aqueous solution , genetics , enzyme
Background Chitosans of high molecular weights have emerged as efficient nonviral gene delivery systems, but the properties and efficiency of well‐defined low molecular weight chitosans (<5 kDa) have not been studied. We therefore characterized DNA complexes of such low molecular weight chitosans and related their physical shape and stability to their efficiency as gene delivery systems in vitro and in vivo . Methods Individual complexes between six different chitosan oligomers (6‐, 8‐, 10‐, 12‐, 14‐ and 24‐mers) and fluorescence‐labeled T4 DNA were visualized and classified into six physical shapes using video‐enhanced fluorescence microscopy. The effects of chitosan chain length, charge ratio (+/−) and solvent properties (pH and ionic strength) on the stability and structure of the complexes were studied. Gene expression in vitro and in vivo were studied using a luciferase reporter gene. Results Free DNA appeared as extended coils. Chitosan complexes had a variety of physical shapes depending on the experimental conditions. In general, the fraction of complexes that had nonaggregated, globular structures increased with increasing chain length of the chitosan oligomer, increasing charge ratio and reduction of pH (from 6.5 to 3.5). A further increase in charge ratio for globular complexes or a further reduction in pH (to 2.5) increased the fraction of aggregates, indicating a window where pharmaceutically desirable globules are obtained. Gene transfection efficiencies in vitro and in vivo were related to the physical shape and stability of the complexes. Only the 24‐mer formed stable complexes that gave a high level of gene expression comparable to that of high molecular weight ultrapure chitosan (UPC) in vitro and in vivo . Conclusions Chitosan oligomers form complexes with DNA in a structure‐dependent manner. We conclude that the 24‐mer, which has more desirable physical properties than UPC, is more attractive as a gene delivery system than the conventional high molecular weight chitosans. Copyright © 2002 John Wiley & Sons, Ltd.