Background: Limited insights into intestinal bacteria's role in diseases like inflammatory bowel disease (IBD) have been due to a scarcity of effective canine in vitro models. Recent advances in Gut-on-a-Chip technology offer new possibilities for studying the interaction between intestinal bacteria and epithelium.
Objectives: To create a microfluidic Gut-on-a-Chip co-culture system with organoids from both healthy and IBD-afflicted dogs.Animals and
Methods: Colonoids from two healthy and two IBD-affected dogs were cultured on the Gut-on-a-Chip, forming villus-like structures under dynamic conditions. We established co-cultures with non-pathogenic Escherichia coli (NPE) and compared them with static Transwell (TW) co-cultures. We assessed epithelial barrier integrity by measuring transepithelial electrical resistance (TEER) and used immunofluorescence staining (IF) to evaluate ZO-1, a tight junction protein, expression.
Results: The Gut-on-a-Chip models, after 6-9 days, showed 3D morphogenesis in the form of villus-like structures in both healthy and IBD dog-derived cells. IF confirmed ZO-1 presence, indicating tight junction formation. NPE infection in the TW system significantly reduced TEER (48.2±4.4% at 12 hours and 7.3±5.3% at 24 hours), whereas the Gut-on-a-Chip maintained TEER levels (100.3±12.8% at 12 hours, 94.9±2.2% at 24 hours, and 88.8±15.0% at 48 hours), preserving 3D structure.Conclusions and clinical importance: The canine Gut-on-a-Chip model successfully replicates villus-like structures and maintains a 3D epithelial configuration with both healthy and IBD organoids. This platform is a significant step forward for studying bacterial-epithelial interactions in veterinary and human medicine, offering a dynamic system for co-culturing live bacteria and preserving epithelial integrity.
