Probing the Mechano-Redox Control of Cell Movement Using Microfluidic Assays.

The mechano-redox phenomenon-where mechanical forces and redox reactions synergistically regulate protein function-has been demonstrated in several mammalian proteins. Integrins, which mediate cellular adhesion to ligands, are particularly sensitive to redox modulation via thiol-disulfide exchange catalyzed by enzymes such as thiol isomerases. These redox-driven modifications can alter integrin conformation and ligand-binding affinity, ultimately leading to dynamic regulation of cell adhesion and de-adhesion. In this work, we present a suite of functional assays designed to investigate how manipulation of thiol exchange reactions influences key cellular processes, including thrombosis and inflammation. These assays employ either thiol-reactive enzymes or small-molecule inhibitors to modulate redox states. They include: (1) a cellular de-adhesion assay to monitor integrin-ligand interactions, (2) a neutrophil crawling assay to assess integrin-mediated motility, (3) a microfluidic thrombus formation assay to evaluate the contribution of thiol isomerases to clot development, (4) the "endo-chip"-a microfluidic platform integrating an endothelium for modelling vascular thromboinflammatory responses under flow, and (5) the microvascular-on-a-post chip, an endothelialized microfluidic device with fibrin-rich thrombosis localized on a 3D post.