Porter Annie, Fan Songshan, Peng Ying, Lv Mengxi, Zhou Yilu, Alanazi Abdulaziz, Han Lin, Wang Liyun, Lu X Lucas
Department of Mechanical Engineering, University of Delaware, Newark, Delaware, 19716, USA.
School of Biomedical Engineering, Science and Health Systems, Drexel University, Philadelphia, PA 19104, USA.
bioRxiv. 2025 May 14:2025.05.08.652928. doi: 10.1101/2025.05.08.652928.
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 . 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.
本研究提出了一种灵敏且经济高效的方法，利用无铜点击化学反应对新的细胞外基质（ECM）成分进行荧光标记，以量化ECM的合成与降解。该方法能够对特定ECM周转进行空间可视化和纵向测量。我们在多个平台上验证了该方法，包括天然软骨外植体以及人间充质干细胞和乳腺癌细胞的单层培养。通过检测减轻炎症诱导的ECM降解的化合物，该技术在骨关节炎药物筛选中也被证明是有效的。与传统的生化或组织学检测相比，这种基于点击化学的技术具有更高的灵敏度、更低的样本需求和更好的时间分辨率。其多功能性支持在组织工程、再生医学、疾病建模和高通量药物评估中的广泛应用。
