In vivo release and gene expression of plasmid DNA by hydrogels of gelatin with different cationization extents
In vivo release and gene expression of plasmid DNA by hydrogels of gelatin with different cationization extents
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DOI：
10.1016/s0168-3659(03)00197-4
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发表时间：
2003-06-24
影响因子：
10.8
通讯作者：
Tabata, Y
中科院分区：
文献类型：
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作者：
Kushibiki, T;Tomoshige, R;Tabata, Y
研究了阳离子明胶水凝胶对lacZ质粒DNA（pSV-lacZ）的体内释放和基因表达的影响。采用水溶性碳二亚胺对明胶羧基进行胺化，通过改变乙二胺的用量，制备了不同阳离子化程度的明胶。将制备的阳离子化明胶与不同浓度的戊二醛（GA）交联，得到不同阳离子化程度的阳离子化明胶水凝胶，作为质粒DNA的释放载体。当将I-125标记的pSV-lacZ的阳离子化明胶水凝胶植入小鼠的股肌中时，剩余的放射性随时间降低，并且保留期随着用于水凝胶制备的GA浓度的增加而延长。用I-125标记的阳离子化明胶水凝胶的体内实验显示，GA浓度越高，每种阳离子化明胶水凝胶的放射性剩余的体内保留期越长。仅对于由胺化百分比为29.7、41.6和47.8摩尔%的明胶制备的水凝胶，pSV-lacZ保留的时间曲线与水凝胶保留的时间曲线良好相关。掺入pSV-lacZ的阳离子化明胶水凝胶的基因表达依赖于明胶的胺化百分比，并且在41.6mol.%或更高. pSV-lacZ可能与阳离子化明胶的降解片段复合并释放正电荷，导致基因表达增强。我们得出结论，明胶与阳离子化程度至少为41.6摩尔%，是通过水凝胶释放系统增强质粒DNA的体内基因表达所需要的。(C)2003 Elsevier Science B. V.保留所有权利。
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 I-125-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 I-125-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. (C) 2003 Elsevier Science B.V. All rights reserved.