biostudies-literatureUnknown11Santoni M<h4>Introduction</h4>Current <i>in vitro</i> intestinal models lack the mechanical forces present in the physiological environment, limiting their reliability for nanotoxicology studies. Here, we developed an enhanced Caco-2/HT29-MTX-E12 co-culture model incorporating orbital mechanical stimulation to better replicate intestinal conditions and investigate nanoparticle interactions.<h4>Methods</h4>We established co-cultures under static and dynamic conditions, evaluating their development through multiple approaches including barrier integrity measurements, gene expression analysis, and confocal microscopy. We introduced novel quantitative analysis of dome formation as a differentiation marker and demonstrated the model application by investigating cellular responses to titanium dioxide (TiFrontiers in molecular biosciences1529027https://www.ebi.ac.uk/biostudies/studies/S-EPMC11769796biostudies-literatureEnhanced intestinal epithelial co-culture model with orbital mechanical stimulation: a proof-of-concept application in food nanotoxicology.PMC11769796Santoni MRandi MRLiguori GMilani LDanesi FPiccinini GBaroncini MfalseEnhanced intestinal epithelial co-culture model with orbital mechanical stimulation: a proof-of-concept application in food nanotoxicology.<h4>Introduction</h4>Current <i>in vitro</i> intestinal models lack the mechanical forces present in the physiological environment, limiting their reliability for nanotoxicology studies. Here, we developed an enhanced Caco-2/HT29-MTX-E12 co-culture model incorporating orbital mechanical stimulation to better replicate intestinal conditions and investigate nanoparticle interactions.<h4>Methods</h4>We established co-cultures under static and dynamic conditions, evaluating their development through multiple approaches including barrier integrity measurements, gene expression analysis, and confocal microscopy. We introduced novel quantitative analysis of dome formation as a differentiation marker and demonstrated the model application by investigating cellular responses to titanium dioxide (Ti2024-01-01T00:00:00Z20242025-04-04T14:05:37.753Z2025-04-04T14:05:37.753ZS-EPMC117697963987216510.3389/fmolb.2024.1529027