Dose and route dependent effects of the mycotoxin deoxynivalenol in a 3D gut-on-a-chip model with flow

Abstract

Deoxynivalenol (DON) is the most prevalent mycotoxin in human food and is ubiquitously detected in human bodyfluids. DON leads to intestinal barrier dysfunction, as observed from animal- and cell culture models with the known disadvantages. Here we present the effects of DON in a gut-on-a-chip model, as the first study incorporating the effects of intestinal flow. Using the OrganoPlate 3-lane, Caco-2 cells were seeded against an extracellular matrix (ECM) and formed leak tight tubules. DON was then applied in different concentrations (3 μM to 300 μM) via the apical or the basolateral channel. Permeability was assessed using continuous TEER and barrier integrity assays (BIA). Zonulin-1, toxicity (LDH) and proinflammatory status (IL-8) was analyzed. DON exposure led to a dose dependent decrease in para-and transcellular barrier integrity, which was more sensitive to basal than apical application (route). Timelaps/Continuous TEER measurements however revealed bidirectional effects, with even TEER-inducing effects of lower concentrations (until 10 μM). IL-8 secretion into luminal supernatants was only induced by apical DON. Attributed to the flow, the barrier-disintegrating effects of DON start at higher concentrations than in other culture models. The barrier was more sensitive to basolateral DON, even though DON had to pass the ECM; and IL-8 secretion was independent of TEER-alterations. Thus, the gut-on-a chip model might be a good alternative to further characterize the bidirectional effects of DON with reasonable throughput incorporating flow.