A closed-loop microfluidic platform for enhanced gut organoid maturation via physicochemical control and morphodynamic feedback

Abstract

Organoid technology has fundamentally transformed biomedical research, but remains limited by its inability to manipulate the in vivo physicochemical microenvironment. Here, we introduce a platform that bridges engineering and biology by elevating organoid-on-chip culture from a static process to a dynamic, precise science. We present a closed-loop microfluidic system that integrates on-demand CO₂ microreactions for spatiotemporal control of pH and oxygen, along with a novel suite of label-free, image-based morphodynamic sensors. These sensors include the dynamic deformation index (DDI), which quantifies structural stability; the curvature growth indicator (CGI), which tracks morphogenic activity; and the time-integrated shape entropy (TISE), which measures phenotypic variability. Together, the devices monitor the organoid’s state in real time and autonomously maintain physiological homeostasis. We demonstrate that this approach improves organoid health, achieving over 95% viability, reducing hypoxic core formation by 50% compared to static cultures, and enabling robust, physiologically relevant differentiation. Our versatile toolkit enables precise organoid interrogation and control, sets a new standard for intelligent in vitro systems, with significant implications for accelerating the fidelity of disease modelling, drug screening, and personalised medicine.