title: Atomic force microscopy as analytical tool to study physico-mechanical properties of intestinal cells.
abstract: The small intestine is a complex system that carries out various functions. The main function of enterocytes is absorption of nutrients, whereas membranous cells (M cells) are responsible for delivering antigens/foreign substances to the mucosal lymphoid tissues. However, to get a fundamental understanding of how cellular structures contribute to physiological processes, precise knowledge about surface morphologies, cytoskeleton organizations and biomechanical properties is necessary. Atomic force microscopy (AFM) was used here as a powerful tool to study surface topographies of Caco-2 cells and M cells. Furthermore, cell elasticity (i.e., the mechanical response of a cell on a tip indentation), was elucidated by force curve measurements. Besides elasticity, adhesion was evaluated by recording the attraction and repulsion forces between the tip and the cell surface. Organization of F-actin networks were investigated via phalloidin labeling and visualization was performed with confocal laser scanning fluorescence microscopy (CLSM) and scanning electron microscopy (SEM). The results of these various experimental techniques revealed significant differences in the cytoskeleton/microvilli arrangements and F-actin organization. Caco-2 cells displayed densely packed F-actin bundles covering the entire cell surface, indicating the formation of a well-differentiated brush border. In contrast, in M cells actins were arranged as short and/or truncated thin villi, only available at the cell edge. The elasticity of M cells was 1.7-fold higher compared to Caco-2 cells and increased significantly from the cell periphery to the nuclear region. Since elasticity can be directly linked to cell adhesion, M cells showed higher adhesion forces than Caco-2 cells. The combination of distinct experimental techniques shows that morphological differences between Caco-2 cells and M cells correlate with mechanical cell properties and provide useful information to understand physiological processes/mechanisms in the small intestine.
authors.affiliation: Institute of Pharmaceutical Sciences, Department of Pharmaceutical Technology, NAWI Graz, Karl-Franzens-University of Graz, BioTechMed-Graz, Austria.
authors.forename: Christa
authors.initials: C
authors.lastname: Schimpel
authors.affiliation: Institute of Pharmaceutical Sciences, Department of Pharmaceutical Technology, NAWI Graz, Karl-Franzens-University of Graz, BioTechMed-Graz, Austria.
authors.forename: Oliver
authors.initials: O
authors.lastname: Werzer
authors.affiliation: Medical University of Graz, Center for Medical Research, BioTechMed-Graz, Austria.
authors.forename: Eleonore
authors.initials: E
authors.lastname: Fröhlich
authors.affiliation: Research Unit Electron Microscopic Techniques, Institute of Cell Biology, Histology and Embryology, Center for Medical Research, Medical University of Graz, BioTechMed-Graz, Austria.
authors.forename: Gerd
authors.initials: G
authors.lastname: Leitinger
authors.affiliation: Medical University of Graz, Center for Medical Research, BioTechMed-Graz, Austria.
authors.forename: Markus
authors.initials: M
authors.lastname: Absenger-Novak
authors.affiliation: Institute of Pharmaceutical Sciences, Department of Pharmaceutical Technology, NAWI Graz, Karl-Franzens-University of Graz, BioTechMed-Graz, Austria.
authors.forename: Birgit
authors.initials: B
authors.lastname: Teubl
authors.affiliation: Institute of Pharmaceutical Sciences, Department of Pharmaceutical Technology, NAWI Graz, Karl-Franzens-University of Graz, BioTechMed-Graz, Austria.
authors.forename: Andreas
authors.initials: A
authors.lastname: Zimmer
authors.affiliation: Institute of Pharmaceutical Sciences, Department of Pharmaceutical Technology, NAWI Graz, Karl-Franzens-University of Graz, BioTechMed-Graz, Austria ; Research Center Pharmaceutical Engineering GmbH, Graz, Austria.
authors.forename: Eva
authors.initials: E
authors.lastname: Roblegg
keywords: Caco-2 cells; M cells; atomic force microscopy; elasticity; mechanical properties