Human exposure to foodborne inorganic nanoparticles (NPs) is a growing concern. However, identifying potential hazards linked to NP ingestion often requires long-term exposure in animals. Owing these constraints, intestinal organoids are a promising alternative to in vivo experiments, as such an in vitro approach should enable rapid and reliable assessment of the effects on the gut of ingested chemicals. However, this remains to be validated for inorganic substances. In our study, a transcriptomic analysis and immunofluorescence staining were performed to compare the effects of food-grade TiO2 (fg-TiO2) on enteroid-derived monolayers (EDMs) from murine intestinal organoids to known impacts of TiO2 on intestinal epithelium. After validating their ability to dose-dependently respond to a pro-inflammatory cytokine cocktail, EDMs were exposed to 0.1, 1 or 10 µg fg-TiO2/mL for 24h. In line with known data, a dose-related increase of muc2, vilin 1 and chromogranin A gene markers of cell differentiation was observed. In addition, fg-TiO2 induced apoptosis and dose-dependent genotoxicity, while a decreased expression of genes encoding for antimicrobial peptides, and of genes related to tight junction function was observed. These results validated the use of EDMs as a reliable model for toxicity testing of foodborne NPs likely to affect the intestinal barrier.