Methylated N-(4-N,N-dimethylaminobenzyl) chitosan as effective gene carriers: Effect of degree of substitution
Methylated N-(4-N,N-dimethylaminobenzyl) chitosan as effective gene carriers: Effect of degree of substitution
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DOI：
10.1016/j.carbpol.2008.07.012
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发表时间：
2009-01
影响因子：
11.2
通讯作者：
P. Opanasopit;Maleenart Petchsangsai;T. Rojanarata;T. Ngawhirunpat;W. Sajomsang;U. Ruktanonchai
中科院分区：
文献类型：
--
作者：
P. Opanasopit;Maleenart Petchsangsai;T. Rojanarata;T. Ngawhirunpat;W. Sajomsang;U. Ruktanonchai
本研究的目的是研究季节化N-(4-N，N-二甲氨基苯甲基)壳聚糖(TM-BZ-CS)在人肝癌细胞株Huh7上的转染率。考察了季胺化度(DQ)、二甲氨基苯取代度(DS)和聚合物/DNA重量比等因素对转染率的影响。结果表明，所有TM-BZ-CS衍生物均能与DNA缩合。琼脂糖凝胶电泳法显示，当TM57-Bz42-CS/DNA的质量比大于0.5时，TM57-Bz42-CS/DNA和TM57-Bz17-CS/DNA形成了完整的复合体。壳聚糖衍生物的转染率依次为TM57-Bz42-CS>、TM47-Bz42-CS>、TM57-Bz18-CS。培养液pH值对TM57-Bz42-CS/DNA复合体的转染率无影响，但对壳聚糖/DNA复合体的转染率有影响。结果表明，由于壳聚糖表面的疏水基团(N，N-二甲氨基苯甲基)取代了壳聚糖，促进了与DNA的相互作用和缩合，以及N-季胺化，从而增加了壳聚糖的水溶性，增强了基因的表达，从而提高了基因的转染率。在细胞毒性研究中，TM-BZ-CS在最高转染量时是安全的。总之，这种新型的壳聚糖衍生物TM57-Bz42-CS通过有效的DNA缩合和介导最高水平的基因在Huh7细胞中的细胞毒性而显示出作为基因载体的潜力。
The objective of this study was to investigate the transfection efficiency of quaternized N-(4-N,N-dimethylaminobenzyl) chitosan; TM-Bz-CS, using the plasmid DNA encoding green fluorescent protein (pEGFP-C2) on human hepatoma cell lines (Huh7 cells). The factors affecting the transfection efficiency e.g. degree of quaternization (DQ), the degree of dimethylaminobenzyl substitution (DS) and polymer/DNA weight ratio, have been evaluated. The results revealed that all TM-Bz-CS derivatives were able to condense with DNA. Illustrated by agarose gel electrophoresis, complete complexes of TM57-Bz42-CS/DNA were formed at weight ratio of above 0.5, whereas those of TM47-Bz42-CS/DNA and TM57- Bz17-CS/DNA were above 1. The rank of transfection efficiency of the chitosan derivatives were TM57-Bz42-CS>TM47-Bz42-CS>TM57-Bz18-CS. The pH of culture medium did not affect the transfection efficiency of TM57-Bz42-CS/DNA complex, whereas it affected the transfection efficiency of chitosan/DNA complex. The results indicated that the improved gene transfection was due to the hydrophobic group (N,N-dimethylaminobenzyl) substitution on chitosan which promoted the interaction and condensation with DNA as well as N-quaternization which increased chitosan water solubility and enhance gene expression. For cytotoxicity studies, TM-Bz-CS was safe at the concentration of the highest transfection. In conclusion, this novel chitosan derivative, TM57-Bz42-CS showed elevated potential as gene carrier by efficient DNA condensation and mediated highest level of gene transfection with negligible cytotoxicity in Huh7 cells.