Collagen‐Based Gut‐on‐Chip for In Vitro Modeling of Intestinal Barrier Function and Host‐Pathogen Interactions

ABSTRACT The human intestine, characterized by villi and crypt structures, is central to nutrient absorption, barrier function, and host defense. Traditional in vitro models using synthetic membranes such as polydimethylsiloxane (PDMS) and polycarbonate (PC) often fail to accurately replicate the complex intestinal environment. To overcome this, we developed a gut‐on‐chip (GOC), incorporating a thin, porous collagen type I membrane, to better mimic the natural extracellular matrix (ECM) and create a more physiologically relevant system. Membranes were fabricated and characterized by profilometry to determine their thickness, which closely approximates the in vivo intestinal basement membrane. Caco‐2 cells cultured within the device exhibit villi‐like structures, form tight junctions, produce mucin, and establish a functional barrier, demonstrating rapid and stable differentiation on the collagen membrane. We further investigated the device's capacity to model host‐pathogen interactions by infecting the epithelial layer with Candida albicans . Confocal microscopy reveals hyphal invasion of the epithelial cells, while permeability assays demonstrate increased barrier permeability, highlighting the device's ability to replicate infection processes and their impact on barrier integrity. By integrating a physiological membrane and replicating key structural and functional features of the intestine, this GOC provides a promising biomimetic platform for studying intestinal physiology and host‐pathogen dynamics.