Abstract
Inflammatory cytokines and proteins are essential indicators of immune status and disease progression; however, conventional assays rely on invasive sampling and complex processing, restricting their use in real-time monitoring. Here, we present a 3D-printed poly(methyl methacrylate) (PMMA) microneedle-based biosensing platform integrated with a tyramide signal amplification–enhanced enzyme-linked immunosorbent assay (TSA–ELISA) for noninvasive and highly sensitive detection of inflammatory biomarkers in interstitial fluid. The microneedles exhibit precise geometry, adequate mechanical strength, and excellent biocompatibility, facilitating efficient skin penetration and biomarker capture. Stepwise chemical functionalization ensured stable antibody immobilization, while TSA significantly amplified detection signals. The platform achieved reliable, reproducible, and multiplex detection of cytokines and albumin in both healthy individuals and patients with inflammatory skin conditions. Notably, the measured cytokine level in lesional skin of eczema patients was 97.7 pg/mL, showing a significant difference from the 62.8 pg/mL observed in healthy subjects. This MN-based TSA–ELISA system offers a robust and minimally invasive strategy for monitoring inflammation-related biomarkers, holding great potential for clinical diagnostics and personalized healthcare applications.