In vivo release and gene expression of plasmid DNA by hydrogels of gelatin with different cationization extents.
The objective of this paper is to investigate the in vivo release and gene expression of lacZ plasmid DNA (pSV-lacZ) by the hydrogels of cationized gelatin. Gelatin with different cationization extents was prepared by changing the amount of ethylenediamine added to aminize the carboxyl groups of gelatin with a water-soluble carbodiimide. The cationized gelatin prepared was crosslinked by various concentrations of glutaraldehyde (GA) to obtain cationized gelatin hydrogels with different cationization extents as the release carrier of plasmid DNA. When the cationized gelatin hydrogels incorporating 125I-labeled pSV-lacZ were implanted into the femoral muscle of mice, the radioactivity remaining decreased with time and the retention period was prolonged with an increase in the concentration of GA used for hydrogel preparation. In vivo experiments with 125I-labeled cationized gelatin hydrogels revealed that the higher the GA concentration, the longer the in vivo retention period of radioactivity remaining for every cationized gelatin hydrogel. Only for the hydrogels prepared from gelatin with the aminized percentages of 29.7, 41.6, and 47.8 mol.%, the time profile of pSV-lacZ retention correlated well with that of hydrogel retention. The gene expression by the cationized gelatin hydrogels incorporating pSV-lacZ depended on the aminized percentage of gelatin and was significant at the percentage of 41.6 mol.% or higher. It is possible that the pSV-lacZ was complexed with the degraded fragments of cationized gelatin and released with a positive charge, resulting in enhanced gene expression. We conclude that gelatin with a cationization extent of at least 41.6 mol.% is needed for the enhanced in vivo gene expression of plasmid DNA by the hydrogel release system.
Kushibiki
Toshihiro
T
Department of Biomaterials, Institute for Frontier Medical Sciences, Kyoto University, 53 Kawara-cho, Shogoin, Sakyo-ku, Kyoto 606-8507, Japan.
Tomoshige
Ryuji
R
Fukunaka
Yasunori
Y
Kakemi
Masawo
M
Tabata
Yasuhiko
Y
D000818:Animals; D002412:Cations; D005780:Gelatin / Q000737:chemistry / Q000493:pharmacokinetics*; D005786:Gene Expression Regulation / Q000187:drug effects* / Q000502:physiology; D020100:Hydrogels / Q000737:chemistry / Q000493:pharmacokinetics*; D051379:Mice; D018482:Muscle, Skeletal / Q000187:drug effects / Q000378:metabolism; D010957:Plasmids / Q000235:genetics / Q000378:metabolism*
D002412:Cations; D020100:Hydrogels; D005780:Gelatin