In vitro toxicity evaluation of Tritordeum cereal Introduction: Tritordeum is a novel cereal species resulted from the hybridization between durum wheat (Triticum durum) and a wild barley. (Hordeum chilense). It has accreditation as natural crop since obtained through the use of traditional breeding techniques. Tritordeum includes both nutritional quality of barley and technological properties of wheat and for this is largely accepted as novel cereal for the production of pasta, and bakery product. Objective: Tritordeum is of special interest as a candidate low toxic cereal species for patients suffering of wheat related disorder such as. Here we explored the proteome of two Tritordeum lines named Bulel and Aucan and evaluated the biological effects of their prolamin digested samples (PDGs) in human Caco-2 intestinal epithelium cells; Triticum aestivum (Mieti cv), and Triticum durum (Provental cv) were employed as reference controls. Methodology: Prolamins were extracted and preliminary characterized by proteomic analysis. Prolamins were than digested by gastric and pancreatic proteases and the biological effects of digested prolamin (DP) were determined in Caco-2 cells cultured on transwell inserts. When differentiated for 21 days, cells established a polarized epithelial monolayer that provides a physical and biochemical barrier to the passage of ions and molecules. The resultant system was adopted to explore changes in cell viability, monolayer permeability, structural organization of F-actin microfilaments, and ER-stress, triggered by the different DPs. Main findings: the exposure to Mieti- or Provental-DPs disrupted the tight junctions-permeability barrier with a prompt effect detectable after 4h incubation time and wide effects on barrier integrity (TEER) at 24h. Conversely, Aucan-DPs did not enhance monolayer permeability; whereas, Bulel-WDG exerted only slight effects. Moreover, Mieti- and Provental-DPs prompted a noticeable toxicity as confirmed by the reorganization of enterocyte cytoskeleton, in addition to changes in cell viability. Differently, Aucan-DPs and Bulel-DPs neither enhance monolayer viability nor alter the cytoskeleton structure. Proteomic analysis showed that the protein prifiles of Tritordeum lines were sufficiently different from those of common T. durum and T. aestivum to determine a lower in vitro toxicity following gastroduodenal digestion. Conclusion: Outcomes support the use of novel Tritordeum lines as prospective candidates for the development of innovative approaches to reduce wheat flour toxicity.
IN VITRO TOXICITY AND NUTRITIONAL EVALUATION OF TRITORDEUM CEREAL .
IACOMINO G;DANZI D;LAURIA F;MAMONE G
2019
Abstract
In vitro toxicity evaluation of Tritordeum cereal Introduction: Tritordeum is a novel cereal species resulted from the hybridization between durum wheat (Triticum durum) and a wild barley. (Hordeum chilense). It has accreditation as natural crop since obtained through the use of traditional breeding techniques. Tritordeum includes both nutritional quality of barley and technological properties of wheat and for this is largely accepted as novel cereal for the production of pasta, and bakery product. Objective: Tritordeum is of special interest as a candidate low toxic cereal species for patients suffering of wheat related disorder such as. Here we explored the proteome of two Tritordeum lines named Bulel and Aucan and evaluated the biological effects of their prolamin digested samples (PDGs) in human Caco-2 intestinal epithelium cells; Triticum aestivum (Mieti cv), and Triticum durum (Provental cv) were employed as reference controls. Methodology: Prolamins were extracted and preliminary characterized by proteomic analysis. Prolamins were than digested by gastric and pancreatic proteases and the biological effects of digested prolamin (DP) were determined in Caco-2 cells cultured on transwell inserts. When differentiated for 21 days, cells established a polarized epithelial monolayer that provides a physical and biochemical barrier to the passage of ions and molecules. The resultant system was adopted to explore changes in cell viability, monolayer permeability, structural organization of F-actin microfilaments, and ER-stress, triggered by the different DPs. Main findings: the exposure to Mieti- or Provental-DPs disrupted the tight junctions-permeability barrier with a prompt effect detectable after 4h incubation time and wide effects on barrier integrity (TEER) at 24h. Conversely, Aucan-DPs did not enhance monolayer permeability; whereas, Bulel-WDG exerted only slight effects. Moreover, Mieti- and Provental-DPs prompted a noticeable toxicity as confirmed by the reorganization of enterocyte cytoskeleton, in addition to changes in cell viability. Differently, Aucan-DPs and Bulel-DPs neither enhance monolayer viability nor alter the cytoskeleton structure. Proteomic analysis showed that the protein prifiles of Tritordeum lines were sufficiently different from those of common T. durum and T. aestivum to determine a lower in vitro toxicity following gastroduodenal digestion. Conclusion: Outcomes support the use of novel Tritordeum lines as prospective candidates for the development of innovative approaches to reduce wheat flour toxicity.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.