Cytoskeleton reorganization and ultrastructural damage induced by gliadin in a three-dimensional in vitro model

doi:10.3748/wjg.v11.i48.7597

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Dec 28, 2005 (publication date) through Apr 18, 2024

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World Journal of Gastroenterology

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1007-9327

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Baishideng Publishing Group Inc, 7041 Koll Center Parkway, Suite 160, Pleasanton, CA 94566, USA

Basic Research

World J Gastroenterol. Dec 28, 2005; 11(48): 7597-7601
Published online Dec 28, 2005. doi: 10.3748/wjg.v11.i48.7597

Cytoskeleton reorganization and ultrastructural damage induced by gliadin in a three-dimensional in vitro model

Ersilia Dolfini, Leda Roncoroni, Luca Elli, Chiara Fumagalli, Roberto Colombo, Simona Ramponi, Fabio Forlani, Maria Teresa Bardella

Ersilia Dolfini, Leda Roncoroni, Chiara Fumagalli, Department of Biology and Genetics for Health Sciences, University of Milan, Milan, Italy

Luca Elli, Maria Teresa Bardella, Department of Digestive Diseases and Endocrinology, IRCCS Ospedale Maggiore di Milano, University of Milan, Milan, Italy

Roberto Colombo, Department of Biology, University of Milan, Milan, Italy

Simona Ramponi, Milan Research Imaging Centre, Bracco SpA, Milan, Italy

Fabio Forlani, Department of Agrifood Molecular Sciences, University of Milan, Milan, Italy

Author contributions: All authors contributed equally to the work.

Supported by the “Fondazione San Paolo” grant to “Centro per lo Studio della Celiachia”

Correspondence to: Dr Ersilia Dolfini, Department of Biology and Genetics for Health Sciences, Via Viotti 3/5, 20131 Milano, Italy. ersilia.dolfini@unimi.it

Telephone: +39-02-50315837 Fax: +39-02-50315864

Received: March 1, 2005
Revised: July 3, 2005
Accepted: July 8, 2005
Published online: December 28, 2005

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