Evaluation of chitosan salts as non-viral gene vectors in CHO-K1 cells

被引：88
|
作者：
Weecharangsan, Wanlop [1 ]
Opanasopit, Praneet [1 ]
Ngawhirunpat, Tanasait [1 ]
Apirakaramwong, Auayporn [1 ]
Rojanarata, Theerasak [1 ]
Ruktanonchai, Uracha [2 ]
Lee, Robert J. [3 ]
机构：
[1] Silpakorn Univ, Fac Pharm, Nanotechnol Drug Gene Delivery Syst Grp, Nakhon Pathom 73000, Thailand
[2] Natl Nanotechnol Ctr, Pathum Thani 12120, Thailand
[3] Ohio State Univ, Coll Pharm, NCI, OSU Comprehens Canc Ctr,NSF Nanoscales Sci & Engn, Columbus, OH 43210 USA
来源：
INTERNATIONAL JOURNAL OF PHARMACEUTICS | 2008年 / 348卷 / 1-2期
关键词：
chitosan salt; transfection efficiency; gene delivery; CHO-K1; cells;
D O I：
10.1016/j.ijpharm.2007.07.011
中图分类号：
R9 [药学];
学科分类号：
1007 ;
摘要：
The aim of this study was to investigate chitosan/DNA complexes formulated with various chitosan salts (CS) including chitosan hydrochloride (CHy), chitosan lactate (CLa), chitosan acetate (CAc), chitosan aspartate (CAs) and chitosan glutamate (CGI). They were assesed for their DNA complexing ability, transfection efficiency in CHO-K1 (Chinese hamster ovary) cells and their effect on cell viability. CHy, CLa, CAc, CAs and CGI, MW 45 kDa formed a complex with pcDNA3-CMV-Luc at various N/P ratios. CGI/DNA complexes were formulated with various chitosan molecular weights (20, 45, 200 and 460 kDa). The CS/DNA complexes were characterized by agarose gel electrophoresis and investigated for their transfection efficiency in CHO-K1 cells. The cytotoxicity of the complexes was determined by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay in CHO-K1 cells. Gel electrophoresis illustrated that complete complexes formed at Nip ratios above 2 in all CS of MW 45 kDa. The transfection efficiency of CS/DNA complexes was dependent on the salt form and MW of chitosan, and the N/P ratio of CS/DNA complexes. Of different CS, the maximum transfection efficiency was found in different N/P ratios. CHy/DNA, CLa/DNA, CAc/DNA, CAs/DNA and CGI/DNA complexes showed maximum transfection efficiencies at N/P ratios of 12, 12, 8, 6 and 6, respectively. Cytotoxicity results showed that all CS/DNA complexes had low cytotoxicity. This study suggests CS have the potential to be used as safe gene delivery vectors. (c) 2007 Elsevier B.V. All rights reserved.
引用
收藏
页码：161 / 168
页数：8
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