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

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