In Vivo Monitoring of Thrombo‐Inflammatory Biomarkers via Molecularly Imprinted Polymer‐Integrated Hydrogel Microneedles

Abstract

ABSTRACT Thrombotic disorders are a leading cause of cardiovascular mortality worldwide; however, real‐time, point‐of‐care monitoring technologies for timely detection of evolving coagulopathies remain inaccessible. Wearable, minimally invasive tracking of thrombo‐inflammatory activity could enable earlier risk assessment and more effective therapy monitoring than conventional episodic blood tests. Here, we present a reagent‐free, wearable electrochemical platform that integrates an on‐chip Prussian Blue (PB) redox transducer with a signal‐off molecularly imprinted polymer (MIP) layer and a biocompatible hydrogel microneedle (HMN) array for interstitial fluid (ISF) sampling, enabling direct electrochemical detection of thrombo‐inflammatory biomarkers. The electrochemical PB/MIP (e‐MIP) biosensor was configured to quantify thrombin (thrombotic biomarker) as well as interleukin‐6 (IL‐6) and tumor necrosis factor‐α (TNF‐α) (inflammatory biomarkers) directly in dermal ISF extracted via the integrated HMNs. The wearable e‐MIP was characterized in vitro and ex vivo, where porcine skin tests preserved linearity and achieved limits of detection (LODs) of 0.26 ng mL − 1 for thrombin and ≤ 0.41 pg mL − 1 for IL‐6 and TNF‐α, confirming sensitive performance in a skin model. Also, selectivity studies against potential interferents (e.g., prothrombin or cardiac troponins) were conducted to assess the possible cross‐reactivity. in vivo, HMN‐integrated patches applied to lipopolysaccharide (LPS)–challenged rats sampled ISF and delivered it to the e‐MIP, which captured the rise‐and‐fall kinetics of thrombin and IL‐6 over 0–24 h. The results were validated against parallel enzyme‐linked immunosorbent assays (ELISAs) performed on plasma collected at corresponding time points. Its versatile architecture and demonstrated in vivo performance position it as a promising platform that can enable early thrombotic risk assessment and therapeutic monitoring, with potential applications in personalized cardiovascular management following clinical validation.