<i>In vitro</i> 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 <i>in vitro</i> liver-on-a-chip platform focused on modelling the space of Disse (sD) - a key microenvironment, yet insufficiently studied, which mediates cross-talk between endothelial and hepatic systems. The system integrates a biocompatible sodium alginate hydrogel whose mechanical properties were optimised 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 showed viability and polarisation for 8 days, forming organised tissues with functional bile canaliculi. The presence of LSECs markedly enhanced hepatic performance, reflected by increased albumin and urea secretion and activation of pro-regenerative secretory pathways. Prolonged acetaminophen exposure demonstrated the model's capacity to reproduce hepatotoxic responses, confirming its predictive relevance. Such versatile microphysiological platform might become a powerful tool for studying endothelial-hepatic communication, modelling liver pathologies, and improving preclinical DILI testing.