In Vitro Space of Disse Model for Exploration of Drug Induced Hepatotoxicity

Abstract

The limited predictive power of animal models remains a major bottleneck in drug development, particularly in assessing drug-induced liver injury (DILI). To address this, we developed a novel in vitro liver-on-a-chip platform focused on modeling the space of Disse (sD)-a critical yet underrepresented microenvironment mediating endothelial-hepatic interactions. The system integrates a biocompatible sodium alginate hydrogel whose mechanical properties were optimized to mimic physiological liver stiffness, enabling molecular cross-talk between human liver sinusoidal endothelial cells (LSECs) and HepaRG hepatocytes without direct contact. Under dynamic perfusion, the co-culture maintained viability and polarization for eight days, forming organized tissues with functional bile canaliculi. The presence of LSECs markedly enhanced hepatic performance, reflected by increased albumin and urea secretion and activation of proregenerative secretory pathways. Proof-of-concept studies with chronic acetaminophen exposure demonstrated the model's capacity to reproduce hepatotoxic responses, confirming its predictive relevance. This versatile and physiologically relevant microphysiological platform offers a powerful tool for studying endothelial-hepatic communication, modeling liver pathologies, and improving preclinical DILI testing. Its modular design enables future integration of Kupffer and stellate cells to simulate immune and fibrotic responses, extending its applicability to complex liver disease modeling.