A vascularized modular 3D in vitro liver model to study bacterial infection and the role of associated host protein markers

Abstract

Engineering physiologically relevant liver models that can replicate infection dynamics and enable real-time biomarker monitoring remains a critical unmet need in liver disease research and diagnostics. Here, we present a modular, vascularized 3D in vitro liver model using core–shell microscaffolds designed to emulate the hepatic sinusoidal architecture and inflammatory microenvironment. The system co-encapsulates Huh-7 hepatocytes and liver sinusoidal endothelial cells (LSECs) with HUVECs in two distinct regions of the scaffold, the core and shell, respectively, promoting endothelial–hepatocyte cross-talk and secretion of regenerative cytokines including HGF, IL-8, and G-CSF. Upon Escherichia coli infection, the model exhibited reduced albumin levels and upregulated expression of key inflammatory markers, replicating infection-induced hepatic stress. To enable temporal biomarker tracking, we integrated the microscaffold model with a label-free, three-electrode electrochemical biosensor for detecting antibody binding changes using square wave and cyclic voltammetry (SWV/CV). This allowed real-time, non-invasive quantification of IL-6, CRP, and PCT over two weeks, with high sensitivity, which was further corroborated by the standard detection method of ELISA. This bioengineered platform uniquely combines a tissue-mimetic structure, regenerative biology, and real-time biosensing, offering a powerful tool for infection modelling, sepsis diagnostics, and therapeutic screening. Its modularity and scalability position it as a bioengineered platform for studying infection dynamics and enabling pre-clinical therapeutic screening in a physiologically relevant 3D microenvironment.