Bread wheat gliadin cytotoxicity: a new three-dimensional cell model.
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 microg/mL) for 7 days in order to evaluate cell viability (colony-forming assay), and at the standard concentration of 500 microg/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.


CONCLUSIONS
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.
Dolfini
E
E
Department of Biology and Genetics, University of Milan, Italy.
Elli
L
L
Ferrero
S
S
Braidotti
P
P
Roncoroni
L
L
Dasdia
T
T
Falini
M L
ML
Forlani
F
F
Bardella
M T
MT
D000230:Adenocarcinoma / Q000473:pathology; D002446:Celiac Disease / Q000209:etiology* / Q000473:pathology; D045744:Cell Line, Tumor / Q000187:drug effects; D048429:Cell Size / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D003110:Colonic Neoplasms / Q000473:pathology; D004305:Dose-Response Relationship, Drug; D005903:Gliadin / Q000633:toxicity*; D006801:Humans; D014411:Neoplastic Stem Cells; D018874:Spheroids, Cellular / Q000187:drug effects* / Q000473:pathology / Q000648:ultrastructure
D005903:Gliadin