Effect of Mechanical Agitation on Cationic Liposome Transport across an Unstirred Water Layer in Caco-2 Cells

• Published in: Biological & pharmaceutical bulletin 2016/08/01, Vol.39(8), pp.1293-1299
• Main Authors: Kono, Yusuke, Iwasaki, Ayu, Matsuoka, Kenta, Fujita, Takuya
• Format: Article
• Language: English
• Subjects: Animals; Caco-2 cell; Caco-2 Cells; cationic liposome; Cations; DNA - administration & dosage; Dogs; gene delivery; Gene Expression; Genes, fos - genetics; Humans; Liposomes - administration & dosage; Madin Darby Canine Kidney Cells; mechanical agitation; Plasmids; Proto-Oncogene Proteins c-jun - genetics; RNA, Messenger - metabolism; Transcription Factor RelA - metabolism; Transfection - methods; unstirred water layer; Water
• ISSN: 0918-6158; 1347-5215
• DOI: 10.1248/bpb.b16-00050

To develop an effective oral delivery system for plasmid DNA (pDNA) using cationic liposomes, it is necessary to clarify the characteristics of uptake and transport of cationic liposome/pDNA complexes into the intestinal epithelium. In particular, evaluation of the involvement of an unstirred water layer (UWL), which is a considerable permeability barrier, in cationic liposome transport is very important. Here, we investigated the effects of a UWL on the transfection efficiency of cationic liposome/pDNA complexes into a Caco-2 cell monolayer. When Caco-2 cells were transfected with cationic liposome/pDNA complexes in shaking cultures to reduce the thickness of the UWL, gene expression was significantly higher in Caco-2 cells compared with static cultures. We also found that this enhancement of gene expression by shaking was not attributable to activation of transcription factors such as activator protein-1 and nuclear factor-kappaB (NF-κB). In addition, the increase in gene expression by mechanical agitation was observed at all charge ratios (1.5, 2.3, 3.1, 4.5) of cationic liposome/pDNA complexes. Transport experiments using Transwells demonstrated that mechanical agitation increased the uptake of cationic liposome/pDNA complexes by Caco-2 cells, whereas transport of the complexes across a Caco-2 cell monolayer did not occurr. Moreover, the augmentation of the gene expression of cationic liposome/pDNA complexes by shaking was observed in Madin–Darby canine kidney cells. These results indicate that a UWL greatly affects the uptake and transfection efficiency of cationic liposome/pDNA complexes into an epithelial monolayer in vitro.