Electrochemical Detection of Multianalyte Biomarkers in Wound Healing Efficacy

The targeted diagnosis and effective treatments of chronic skin wounds remain a healthcare burden, requiring the development of sensors for real-time monitoring of wound healing activity. Herein, we describe an adaptable method for the fabrication of carbon ultramicroelectrode arrays (CUAs) on flexible substrates with the goal to utilize this sensor as a wearable device to monitor chronic wounds. As a proof-of-concept study, we demonstrate the electrochemical detection of three electroactive analytes as biomarkers for wound healing state in simulated wound media on flexible CUAs. Notably, to follow pathogenic responses, we characterize analytical figures of merit for identification and monitoring of bacterial warfare toxin pyocyanin (PYO) secreted by the opportunistic human pathogen Pseudomonas aeruginosa. We also demonstrate the detection of uric acid (UA) and nitric oxide (NO^•), which are signaling molecules indicative of wound healing and immune responses, respectively. The electrochemically determined limit of detection (LOD) and linear dynamic range (LDR) for PYO, UA, and NO^• fall within the clinically relevant concentrations. Additionally, we demonstrate the successful use of flexible CUAs for quantitative, electrochemical detection of PYO from P. aeruginosa strains and cellular NO^• from immune cells in the wound matrix. Moreover, we present an electrochemical examination of the interaction between PYO and NO^•, providing insight into pathogen-host responses. Finally, the effects of the antimicrobial agent, silver (Ag⁺), on P. aeruginosa PYO production rates are investigated on flexible CUAs. Our electrochemical results show that the addition of Ag⁺ to P. aeruginosa in wound simulant decreases PYO secretion rates.