Investigating the role of shear stress and mucus in intestinal epithelium homeostasis using computational and organoid-based microphysiological systems

The intestinal epithelium is organized into crypt-villus structures, which support compartmentalized functions such as nutrient absorption and barrier integrity through tightly regulated processes of proliferation, migration, differentiation, and extrusion. The tissue is subjected to mechanical inputs, including shear stress and substrate stiffness, yet the influence of these on epithelial homeostasis and force distribution along the crypt-villus axis remains poorly understood. While in vivo studies offer insights, the complexity of the intestinal environment necessitates robust in vitro models to probe these interactions under controlled conditions. To address this, we developed an in-silico model to simulate the shear stress distribution along the epithelium and the mechanical role of mucus in protecting the epithelium. Our results indicated an increasing shear stress profile from crypt to villus. Building on this, we engineered a perfusable epithelium-on-a-chip platform featuring stiffness-tuned polyacrylamide gels and open-lumen organoid monolayers. The system recapitulates the crypt-villus architecture, permits direct access to the apical surface, and enables precise shear stress modulation. Our experiments demonstrated that shear stress selectively enhances crypt density and extrusion rates while maintaining traction forces and division rates, approaching a homeostatic cell turnover balance. Beyond its immediate findings, this platform offers a versatile tool for exploring epithelial biology, enabling the study of microbial interactions, drug delivery, and disease pathology in physiologically relevant conditions. By bridging the gap between static in vitro systems and the complexity of in vivo tissues, this work provides a foundation for future advances in modelling intestinal systems with precise environmental control.

Funder

Science Foundation Ireland

Publisher

University of Galway

Rights

Attribution-NonCommercial-NoDerivatives 4.0 International

cbn

Collections

University of Galway Theses (PhD Theses)