Click chemistry-based quantification of extracellular matrix turnover for drug screening and regenerative medicine

Abstract

This study presents a sensitive and cost-efficient method to quantify extracellular matrix (ECM) synthesis and degradation using copper-free click chemistry reactions to fluorescently label new ECM components. The approach enables spatial visualization and longitudinal measurement of specific ECM turnover in vitro. We validated the method across multiple platforms, including native cartilage explants and monolayer cultures of human mesenchymal stem cells and breast cancer cells. The technique also proved effective for osteoarthritis drug screening by detecting compounds that mitigate inflammation-induced ECM degradation. Compared to traditional biochemical or histological assays, this click chemistry-based technique offers higher sensitivity, lower sample requirements, and improved temporal resolution. Its versatility supports broad applications in tissue engineering, regenerative medicine, disease modeling, and high-throughput drug evaluation.