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Improvement of antisense oligonucleotides delivery using high hydrostatic pressurized lipoplex

Published online by Cambridge University Press:  20 March 2013

Tsuyoshi Kimura ,
Asami Sano ,
Kwangwoo Nam ,
Kazunari Akiyoshi ,
Yoshihiro Sasaki  and
Akio Kishida
Tsuyoshi Kimura
Affiliation:
Institute of Biomaterials and Bioengineering, Tokyo Medical and Dental University, 2-3-10 Kanda-surugadai, Chiyoda-ku Tokyo 101-0062, Japan
Asami Sano
Affiliation:
Institute of Biomaterials and Bioengineering, Tokyo Medical and Dental University, 2-3-10 Kanda-surugadai, Chiyoda-ku Tokyo 101-0062, Japan
Kwangwoo Nam
Affiliation:
Institute of Biomaterials and Bioengineering, Tokyo Medical and Dental University, 2-3-10 Kanda-surugadai, Chiyoda-ku Tokyo 101-0062, Japan
Kazunari Akiyoshi
Affiliation:
Department of Polymer Chemistry, Kyoto University, Kyoto daigaku-Katsura, Nishikyo-ku, Kyoto 615-8530
Yoshihiro Sasaki
Affiliation:
Institute of Biomaterials and Bioengineering, Tokyo Medical and Dental University, 2-3-10 Kanda-surugadai, Chiyoda-ku Tokyo 101-0062, Japan
Akio Kishida
Affiliation:
Institute of Biomaterials and Bioengineering, Tokyo Medical and Dental University, 2-3-10 Kanda-surugadai, Chiyoda-ku Tokyo 101-0062, Japan
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Abstract

Cationic liposome (CL) is a promising vector for nucleic acid therapy. In the present study, we investigated the effect of high hydrostatic pressure (HHP) treatment to lipoplex on the lipoplex-based antisense oligodeoxynucleotides (AS-ODNs) delivery in order to improve the transfection efficacy of lipoplex. Cationic liposome consisting of DOTMA and DOPE was used. AS-ODNs were designed to inhibit the expression of firefly luciferase. The complexes of CL and AS-ODN were prepared at various C/A ratios and then pressurized hydrostatically at various atmospheres (∼10,000 atm) for 10 min (HHP treatment). After removal of pressure, the pressurized lipoplexes were used. The lipoplex with and without the HHP treatment was transferred into HeLa cells expressing firefly luciferase transiently. The luciferase activity using the HHP-treated lipoplex was decreased compared to that of the non-pressurized lipoplex. Also, for HEK293 cells expressing luciferase stably, the lipoplex with the HHP treatment could effectively suppress the luciferase expression. In order to elucidate relationship between the structure and the transfection efficiency of the HHP-treated lipoplex, the properties of the HHPtreated lipoplex were examined by various physicochemical analyses. The different physicochemical properties between the lipoplexes with and without HHP treatment were showed, suggesting that the nature of lipoplex was changed by the HHP treatment. We believe that this change of lipoplex properties by the HHP treatment affected the efficiency of gene suppression. This HHP treatment for lipoplex appears to be a promising contribution to gene and oligonucleotide delivery.

Keywords

biomaterialbiomimetic (assembly)biomedical
Type
Articles
Information
MRS Online Proceedings Library (OPL) , Volume 1498: Symposium L – Biomimetic, Bio-inspired and Self-Assembled Materials for Engineered Surfaces and Applications , 2013 , pp. 3 - 8
Copyright
Copyright © Materials Research Society 2013 

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References

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