96-Well Format-Based Liver–Heart-on-a-Chip Platform for Facile and Parallel Testing of Drug-Induced Cardiotoxicity after Liver Metabolism

Abstract

Current preclinical drug evaluation predominantly relies on conventional two-dimensional (2D) cell cultures grown in static environments, which lack dynamic interactions and fail to recapitulate complex microenvironments. These approaches are inefficient and overly simplistic, and often require costly animal models that poorly mimic human physiology. Body-on-a-chip (BoC) technology─comprising robust and flexible three-dimensional (3D) microtissues (MTs)─has emerged to address these limitations. BoC platforms constitute microphysiological systems that mimic human organ physiology and function in vitro, enabling improved toxicity screening and personalized medicine. Here, we present a 96–well format-based microfluidic liver–heart-on-a-chip platform that supports parallel, gravity-driven perfusion to efficiently predict and assess multiorgan drug effects. We demonstrate hepatic metabolism of the cardiotoxic prodrug terfenadine (TFND) into its noncardiotoxic metabolite fexofenadine within liver MTs and the consequent physiological–pathological responses of cardiac MTs. The optimized liver–heart-on-a-chip preserves multitissue functionality, underscores the importance of continuous media circulation, and reveals intertissue interactions relevant to drug-induced cardiotoxicity after liver metabolism.