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Cytoskeleton reorganization and ultrastructural damage induced by gliadin in a three-dimensional in vitro model
Authors:Dolfini Ersilia  Roncoroni Leda  Elli Luca  Fumagalli Chiara  Colombo Roberto  Ramponi Simona  Forlani Fabio  Bardella Maria Teresa
Affiliation:1. Department of Biology and Genetics for Health Sciences, University of Milan, Milan, Italy
2. Department of Digestive Diseases and Endocrinology, IRCCS Ospedale Maggiore di Milano, University of Milan, Milan, Italy
3. Department of Biology, University of Milan, Milan, Italy
4. Milan Research Imaging Centre, Bracco SpA, Milan, Italy
5. Department of Agrifood Molecular Sciences, University of Milan, Milan, Italy
Abstract:AIM: To evaluate the interplay between gliadin and LoVo cells and the direct effect of gliadin on cytoskeletal patterns. METHODS: We treated LoVo multicellular spheroids with digested bread wheat gliadin in order to investigate their morphology and ultrastructure (by means of light microscopy and scanning electron microscopy), and the effect of gliadin on actin (phalloidin fluorescence) and the tight-junction protein occludin and zonula occluden-1. RESULTS: The treated spheroids had deep holes and surface blebs, whereas the controls were smoothly surfaced ovoids. The incubation of LoVo spheroids with gliadin decreased the number of intracellular actin filaments, impaired and disassembled the integrity of the tight-junction system. CONCLUSION: Our data obtained from an "in vivo-like" polarized culture system confirm the direct noxious effect of gliadin on the cytoskeleton and tight junctions of epithelial cells. Unlike two-dimensional cell culture systems, the use of multicellular spheroids seems to provide a suitable model for studying cell-cell interactions.
Keywords:Celiac disease  Gliadin  Cytoskeleton  Multicellular spheroids
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