A human liver organoids-on-chip for the assessment of drug-induced liver injury.

BACKGROUND: Drug-induced liver injury (DILI) is a leading cause of liver disease. Current drug toxicology screening primarily relies on 2D cell cultures and animal models, which have limitations in predicting clinical hepatotoxicity, leading to the withdrawal of approved drugs, such as Troglitazone and Fialuridine. To address this, we developed a liver organoids-on-chip (LOoC) system that incorporates human liver progenitor cell-derived organoids and an endothelial barrier to create a biomimetic 3D liver model for the accurate prediction of human-relevant drug responses. METHODS: The LOoC platform was constructed using a customized KabellyInsert™ chip manufactured via injection molding with polycarbonate, featuring eight triple-unit inserts sealed with porous PET membranes to form hepatic and vascular compartments. Liver organoids derived from ethically sourced human liver progenitor cells. These organoids were embedded in the hepatic chamber while liver sinusoidal endothelial cells were seeded in the vascular channel, with dynamic flow generated by orbital shaking. Platform validation included immunofluorescence staining for hepatic markers (CYP3A4, CYP2C9, ALB), functional assessments of urea synthesis and albumin secretion via ELISA assays, and drug testing at clinical Cmax-based concentrations. Hepatotoxicity was evaluated using Hy’s Law criteria with ALT/AST/ALP/LDH measurements, while pharmacokinetic parameters were quantified through LC-MS/MS analysis of compartment-specific effluents. RESULTS: The LOoC model demonstrated mature hepatic gene expression and enhanced function stability, including consistent albumin secretion and urea production over a 7-day culture period. For comprehensive hepatotoxicity assessment, we evaluated seven drugs with varying toxicity profiles (0–8 on a hepatotoxicity level scale), including the species-specific hepatotoxins Troglitazone and Fialuridine (FIAU), using human-relevant blood concentration gradients. The LOoC system achieved up to 93% accuracy in phenotypic toxicity classification and 100% accuracy in hepatotoxicity grading based on sensitivity assessments. CONCLUSION: This study established a robust liver organoid-on-chip (LOoC) platform by integrating functional human liver progenitor cell-derived organoids, a biomimetic endothelial barrier with a dynamic fluidic microenvironment. The LOoC simulated key pharmacokinetic parameters (e.g., The half-time and Km values of acetaminophen in LOoC are similar to human data) and identified species-specific hepatotoxicity (representative drugs: Troglitazone and FIAU). Through Cmax-based gradient dosing and Hy’s Law criteria, the platform demonstrated high sensitivity and accuracy in multiparameters hepatotoxicity assessment, supporting its utility for preclinical drug safety evaluation. CLINICAL TRIAL NUMBER: Not applicable. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s40360-025-01074-z.