title: Effect of Mechanical Agitation on Cationic Liposome Transport across an Unstirred Water Layer in Caco-2 Cells.
abstract: 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.
authors.affiliation: Laboratory of Molecular Pharmacokinetics, College of Pharmaceutical Sciences, Ritsumeikan University.
authors.forename: Yusuke
authors.initials: Y
authors.lastname: Kono
authors.forename: Ayu
authors.initials: A
authors.lastname: Iwasaki
authors.forename: Kenta
authors.initials: K
authors.lastname: Matsuoka
authors.forename: Takuya
authors.initials: T
authors.lastname: Fujita
mesh_terms: D000818:Animals; D018938:Caco-2 Cells; D002412:Cations; D004247:DNA / Q000008:administration & dosage*; D004285:Dogs; D015870:Gene Expression; D016762:Genes, fos / Q000235:genetics; D006801:Humans; D008081:Liposomes / Q000008:administration & dosage*; D061985:Madin Darby Canine Kidney Cells; D010957:Plasmids; D016755:Proto-Oncogene Proteins c-jun / Q000235:genetics; D012333:RNA, Messenger / Q000378:metabolism; D051996:Transcription Factor RelA / Q000378:metabolism; D014162:Transfection / Q000379:methods*; D014867:Water
chemical_list: D002412:Cations; D008081:Liposomes; D016755:Proto-Oncogene Proteins c-jun; D012333:RNA, Messenger; D051996:Transcription Factor RelA; D014867:Water; D004247:DNA