A dynamically cultured intestinal epithelial barrier model with metabolomics assessment for evaluating oxidative injury

Abstract

The study of human intestinal diseases, particularly those involving oxidative stress-induced barrier dysfunction, has attracted increasing attention. Traditional studies have relied heavily on animal models and static 2D cell cultures, and recently, intestinal organ-on-a-chip models have emerged as a promising alternative for modeling intestinal pathophysiology in a human-relevant context. In this study, a high-throughput intestinal chip model was developed using double-sided pressure-sensitive adhesive tape and commercial polycarbonate materials. The model was employed to culture the Caco-2 barrier under continuous fluid flow and cyclic mechanical strain which are crucial for mature barrier formation and function. Bright-field and dark-field microscopy showed that the cells formed a tight, continuous barrier layer within the system. Sodium fluorescein permeation experiments demonstrated good permeability, while polymerase chain reaction (PCR) experiments and laser confocal microscopy imaging further confirmed a high degree of epithelial polarization. Additionally, an oxidative damage model was constructed using hydrogen peroxide. Immunofluorescence staining and metabolomics analysis verified that the model exhibited characteristics consistent with oxidative damage in intestinal cells, indicating the successful construction of the oxidative damage model.