期刊
BIOMATERIALS
卷 32, 期 20, 页码 4659-4669出版社
ELSEVIER SCI LTD
DOI: 10.1016/j.biomaterials.2011.03.013
关键词
Bubble lipoplex; US exposure; Gene transfer; Endocytosis; Pro-inflammatory cytokine production
资金
- Ministry of Education, Culture, Sports, Science and Technology of Japan
- Ministry of Health, Labour and Welfare of Japan
- National Institute of Biomedical Innovation (NIBIO)
- Japan Society for the Promotion of Sciences (JSPS)
- Grants-in-Aid for Scientific Research [10J01875] Funding Source: KAKEN
The development of gene transfection methods enhancing the level of gene expression under simple and low-toxic condition is required for gene therapy in clinical. Our group has developed the ultrasound (US)mediated gene transfection method using Man-PEG(2000) bubble lipoplexes, which are US-responsive and mannose-modified gene carriers, and succeeded in obtaining the enhanced gene expression in mannose receptor-expressing cells selectively by the gene transfer using Man-PEG(2000) bubble lipoplexes with US exposure in vitro and in vivo. Here, we investigated pDNA transferring mechanism followed by US exposure to unmodified and Man-PEG(2000) bubble lipoplexes, in particular, focused on US exposure timing. Following investigation of intracellular transferring characteristics, a large amount of pDNA was transferred into the cytoplasm followed by US-mediated destruction of bubble lipoplexes in the gene transfer using both bubble lipoplexes with US exposure. Moreover, the effective gene expression was obtained without TNF-alpha production when US was exposed until 5 min after the addition of bubble lipoplexes. These findings suggest that the gene transfer using unmodified and Man-PEG(2000) bubble lipoplexes with US exposure enables to transfer pDNA into the cytoplasm, and optimized US exposure timing is important to achieve the high level of gene expression and the low level of pro-inflammatory cytokine production. (C) 2011 Elsevier Ltd. All rights reserved.
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