Towards a minimally invasive platform for point-of-care early detection of NAFLD through rapid profiling of MDA and TNF-α with synthetic MIP receptor

Abstract

Non-alcoholic fatty liver disease (NAFLD) is a progressive metabolic disorder strongly associated with oxidative stress, lipid peroxidation, and inflammatory signaling. Despite its growing prevalence, NAFLD diagnosis still relies primarily on invasive liver biopsy, underscoring the need for alternative approaches. This study presents a novel, dual-analyte molecularly imprinted polymer (MIP)-based biosensing platform designed for minimally invasive detection, metabolic profiling and staging of NAFLD using microneedles. The system targets two key biomarkers— malondialdehyde (MDA), a representative product of lipid peroxidation, and tumor necrosis factor-alpha (TNF-α), a pro-inflammatory cytokine linked to hepatocellular injury and fibrosis. In silico molecular docking was employed to determine and optimize monomer–template interactions for selective recognition. Docking of MDA using its crystal structure (PDB ID: 2C9V) with methacrylic acid (MAA), acrylamide (AAm), and hydroxyethyl methacrylate (HEMA) identified HEMA as the most stable and selective monomer. Similarly, docking analysis for TNF-α (PDB ID: 1TNF) revealed MAA as the most energetically favorable functional monomer, ensuring high imprint fidelity through hydrogen bonding and electrostatic interactions. The synthesized MIPs were integrated into an electrochemical sensing interface for rapid, label-free detection of both analytes in biofluids. This dual MIP biosensor demonstrates high sensitivity, molecular specificity, and operational stability, offering a promising route for minimally invasive or non-invasive, realtime monitoring of metabolic and inflammatory biomarkers in NAFLD.