title: Development of an advanced intestinal in vitro triple culture permeability model to study transport of nanoparticles.
abstract: Intestinal epithelial cell culture models, such as Caco-2 cells, are commonly used to assess absorption of drug molecules and transcytosis of nanoparticles across the intestinal mucosa. However, it is known that mucus strongly impacts nanoparticle mobility and that specialized M cells are involved in particulate uptake. Thus, to get a clear understanding of how nanoparticles interact with the intestinal mucosa, in vitro models are necessary that integrate the main cell types. This work aimed at developing an alternative in vitro permeability model based on a triple culture: Caco-2 cells, mucus-secreting goblet cells and M cells. Therefore, Caco-2 cells and mucus-secreting goblet cells were cocultured on Transwells and Raji B cells were added to stimulate differentiation of M cells. The in vitro triple culture model was characterized regarding confluence, integrity, differentiation/expression of M cells and cell surface architecture. Permeability of model drugs and of 50 and 200 nm polystyrene nanoparticles was studied. Data from the in vitro model were compared with ex vivo permeability results (Ussing chambers and porcine intestine) and correlated well. Nanoparticle uptake was size-dependent and strongly impacted by the mucus layer. Moreover, nanoparticle permeability studies clearly demonstrated that particles were capable of penetrating the intestinal barrier mainly via specialized M cells. It can be concluded that goblet cells and M cells strongly impact nanoparticle uptake in the intestine and should thus be integrated in an in vitro permeability model. The presented model will be an efficient tool to study intestinal transcellular uptake of particulate systems.
authors: Institute of Pharmaceutical Sciences, University of Graz , Graz, Austria.
authors: Christa
authors: C
authors: Schimpel
authors: Birgit
authors: B
authors: Teubl
authors: Markus
authors: M
authors: Absenger
authors: Claudia
authors: C
authors: Meindl
authors: Eleonore
authors: E
authors: Fröhlich
authors: Gerd
authors: G
authors: Leitinger
authors: Andreas
authors: A
authors: Zimmer
authors: Eva
authors: E
authors: Roblegg
mesh_terms: D000818:Animals; D001402:B-Lymphocytes / Q000378:metabolism*; D001692:Biological Transport; D018938:Caco-2 Cells; D002463:Cell Membrane Permeability; D018920:Coculture Techniques; D020895:Enterocytes / Q000378:metabolism*; D020397:Goblet Cells / Q000378:metabolism*; D019073:HT29 Cells; D006801:Humans; D066298:In Vitro Techniques; D007413:Intestinal Mucosa / Q000378:metabolism*; D009093:Mucus / Q000378:metabolism*; D053758:Nanoparticles / Q000737:chemistry*; D011137:Polystyrenes / Q000737:chemistry; D013552:Swine
chemical_list: D011137:Polystyrenes
keywords: Caco-2 cells; M cells; goblet cells; in vitro triple culture; intestinal mucosa; nanoparticles