Double-tether extraction from human umbilical vein and dermal microvascular endothelial cells

Multiple tethers are very likely extracted when leukocytes roll on the endothelium under high shear stress. Endothelial cells have been predicted to contribute more significantly to simultaneous tethers and thus to the overall rolling stabilization. We therefore extracted and quantified double tethers from endothelial cells with the micropipette aspiration technique. We show that the constitutive parameters (threshold force (F0) and effective viscosity (etaeff)) for double-tether extraction are twice those for single-tether extraction and are remarkably similar for human neonatal (F0=105+/-5 pN; etaeff=1.0+/-0.1 pN.s/microm) and adult (F0=118+/-13 pN; etaeff=1.3+/-0.2 pN.s/microm) dermal microvascular, and human umbilical vein (F0=99+/-3 pN; etaeff=1.0+/-0.1 pN.s/microm) endothelial cells. Additionally, these parameters are also independent of surface receptor type, cytokine stimulation, and attachment state of the endothelial cell. We also introduce a novel correlation between the cell-substrate contact stress and gap width, with which we can predict the apparent cell-substrate separation range to be 0.01-0.1 microm during leukocyte rolling. With a biomechanical model of leukocyte rolling, we calculate the force history on the receptor-ligand bond during tether extraction and predict maximum stabilization for the double simultaneous tether extraction case.