Background: In an attempt to clarify the role of gliadin toxicity in the pathogenesis of gluten intolerance (celiac disease), previous in vitro studies have been based on two-dimensional human cell cultures, However, the specific morphological and biochemical properties of in vivo tissue are better maintained in three-dimensional cell cultures (multicellular spheroids, MCS). The aim of this study was to develop a three-dimensional in vitro model to investigate the effects of gliadin on epithelial cells and broaden our understanding of the early tissue damage occurring in celiac disease. Methods: The three-dimensionally growing Lovo cell line was exposed to increasing concentrations of peptic-tryptic-digested bread wheat gliadin (from 125 to 1000 μg/mL) for 7 days in order to evaluate cell viability (colony-forming assay), and at the standard concentration of 500 μg/mL for 7 days in order to evaluate MCS diameters, volumes and cell morphology using light and electron microscopy. Results: In comparison with the controls, the cell viability of the gliadin-treated MCS was significantly reduced (20-80%), but there was no difference in size. Various degrees of cell damage (autophagic vacuoles and intra-cytoplasmic lipid-like droplets) were detected by both light and electron microscopy. Conclusion: This is the first study investigating the effects of gliadin on MCS. Lovo MCS seem to be responsive to gliadin exposure, thus confirming previous results obtained using two-dimensional cell cultures. The data suggest that three-dimensional cell cultures may be useful in broadening our understanding of some of the early effects of gliadin peptides on epithelial cells.

Bread wheat gliadin cytotoxicity : a new three-dimensional cell model / E. Dolfini, L. Elli, S. Ferrero, P. Braidotti, L. Roncoroni, T. Dasdia, M.L. Falini, F. Forlani, M.T. Bardella. - In: SCANDINAVIAN JOURNAL OF CLINICAL & LABORATORY INVESTIGATION. - ISSN 0036-5513. - 63:2(2003), pp. 135-141.

Bread wheat gliadin cytotoxicity : a new three-dimensional cell model

E. Dolfini;L. Elli;S. Ferrero;P. Braidotti;L. Roncoroni;M.L. Falini;F. Forlani;M.T. Bardella
2003

Abstract

Background: In an attempt to clarify the role of gliadin toxicity in the pathogenesis of gluten intolerance (celiac disease), previous in vitro studies have been based on two-dimensional human cell cultures, However, the specific morphological and biochemical properties of in vivo tissue are better maintained in three-dimensional cell cultures (multicellular spheroids, MCS). The aim of this study was to develop a three-dimensional in vitro model to investigate the effects of gliadin on epithelial cells and broaden our understanding of the early tissue damage occurring in celiac disease. Methods: The three-dimensionally growing Lovo cell line was exposed to increasing concentrations of peptic-tryptic-digested bread wheat gliadin (from 125 to 1000 μg/mL) for 7 days in order to evaluate cell viability (colony-forming assay), and at the standard concentration of 500 μg/mL for 7 days in order to evaluate MCS diameters, volumes and cell morphology using light and electron microscopy. Results: In comparison with the controls, the cell viability of the gliadin-treated MCS was significantly reduced (20-80%), but there was no difference in size. Various degrees of cell damage (autophagic vacuoles and intra-cytoplasmic lipid-like droplets) were detected by both light and electron microscopy. Conclusion: This is the first study investigating the effects of gliadin on MCS. Lovo MCS seem to be responsive to gliadin exposure, thus confirming previous results obtained using two-dimensional cell cultures. The data suggest that three-dimensional cell cultures may be useful in broadening our understanding of some of the early effects of gliadin peptides on epithelial cells.
Celiac disease; Cell culture; Gluten intolerance; Multicellular spheroids
Settore MED/08 - Anatomia Patologica
Settore BIO/10 - Biochimica
Settore BIO/13 - Biologia Applicata
Settore MED/12 - Gastroenterologia
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Utilizza questo identificativo per citare o creare un link a questo documento: http://hdl.handle.net/2434/483197
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