Mechanically engineered hydrogels for advanced in vitro drug permeability testing

The development of new drugs is limited by the inadequacy of pre-clinical models mimicking the intestinal barrier. The recapitulation of the intestinal microenvironment could play a pivotal role in enhancing the biomimicry and predictivity of in vitro models for drug absorption studies. We engineered mechanically tuned hydrogels to drive intestinal epithelial cell morphogenesis and sub-differentiation to improve permeability assays predictivity. Substrates were characterized to match the Young’s Modulus of different regions of the intestinal extracellular matrix. Caco-2 cell morphology, polarization and sub-differentiation were quantified by immunostaining. Some insights into the mechanotransduction pathways were provided. The in vitro model was adopted to perform proof-of-concept permeability assays by testing insulin as an example drug, providing more physiologically relevant results for assessing drug transport across the intestinal barrier. This approach could enhance the predictivity of drug absorption studies while ensuring easy translatability of validated protocols and comparability with state-of-the-art data.