Automated Microfluidic Platform for Molecular Transport Analysis across Biomimetic Interfaces

ABSTRACT Advancing drug development and disease modeling requires physiologically relevant in vitro systems that accurately reproduce the dynamic and selective transport functions of human tissue barriers. In response to regulatory efforts to reduce animal testing, there is increasing demand for standardized, scalable, and sensor‐integrated microphysiological platforms. Although organ‐on‐chip technologies have improved biological relevance, many remain limited by technical complexity and insufficient sensing, particularly for non‐equilibrium transport processes that characterize barriers such as the renal tubule, intestine, bloodbrain barrier, and placenta. Here, we present a fully automated microfluidic platform for dynamic and quantitative characterization of molecular transport across biomimetic interfaces. The system emulates key biophysical features, including directional flow, controlled gradient formation, and tunable shear stress, using synthetic, cell‐free architectures. Integrated oxygen sensors enable monitoring of physiologically relevant oxygen levels, while embedded flow sensors support closedyloop control of fluid dynamics and solute delivery. Together, these sensing capabilities allow experimental control and time‐resolved analysis of molecular transport under non‐equilibrium conditions. Validated using acellular models, the platform is engineered for future integration with living tissues and molecular sensorization. Proof‐of‐concept studies demonstrate its ability to reproduce key transport dynamics, highlighting its potential for pharmacokinetic modeling, organ‐on‐chip research, and preclinical drug development.