Collagen-Based Gut-on-Chip for in vitro modeling of intestinal barrier function and host-pathogen interactions

Abstract

The human intestine, characterized by its villi and crypt structures, plays a critical role in nutrient absorption, barrier function, and host defense. However, traditional in-vitro models employing synthetic membranes like polydimethylsiloxane (PDMS) and polycarbonate (PC) often fail to accurately replicate the complex physiological environment of the intestine. To address this limitation, we developed a gut-on-chip, incorporating a porous collagen type I membrane, to better mimic the natural extracellular matrix (ECM) and create a more physiologically relevant in vitro system. Thin, porous collagen type I membranes were fabricated and characterized by linear close contact profilometry to determine their thickness, which closely approximated the in vivo intestinal basement membrane. Caco-2 cells cultured within the device exhibited the formation of villi-like structures, tight junction formation, and mucin production, demonstrating successful differentiation and functional barrier formation on the collagen membrane. We investigated the devices capacity to model host-pathogen interactions by infecting the cell layer with Candida albicans. Confocal microscopy revealed hyphal invasion of the epithelial cells, and permeability assays demonstrated increased layer permeability following infection, highlighting the devices ability to replicate infection processes and their impact on barrier integrity. This gut-on-chip, by integrating physiological membrane and replicating key structural and functional aspects of the intestine, offers a promising platform for studying intestinal physiology and host-pathogen dynamics.