Orbital shaker-driven gut-on-a-chip platform for drug-induced permeability and microenvironment studies

Gut-on-a-chip platforms replicate realistic gut microenvironments but face limited adoption due to their complex designs, expensive fabrication, and specialized instrumentation that increases operational complexity. In this study, we present a microfluidic chip insertable into 12-well plates with a unique radial design and a pumpless flow actuation system using an orbital shaker. We use a surface tension-driven hydrogel patterning technique to compartmentalize the chip, enabling co-culture of gut epithelium and vasculature, resulting in leak-proof monolayer tubes. Furthermore, computational fluid dynamic analysis demonstrates bidirectional peristaltic flow induced by the shaker. The platform's physiological relevance is confirmed through the evaluation of cell polarization, tight junction markers and barrier integrity, using high-magnification microscopy and electrical resistance measurements. We also demonstrate the ability of the platform to support live bacterial colonization, simulating host–microbe interactions. The model is validated for drug development by assessing gut and vascular permeability following drug overdose and inflammatory cytokine activation. Additionally, we explore nanoplastic poisoning using nano polyethylene terephthalate (PET) particles, highlighting the gut's role in limiting particle absorption into the bloodstream. The orbital gut-on-a-chip platform offers an accessible, dynamic cell culture system for drug discovery and biomimetic modeling of gut-related disease interactions.