Circulating leukocytes in trafficking to the inflammatory sites, will be first tether to, and then roll on the vascular surface. This event is mediated through specific interaction of P-selectin and P-selectin glycoprotein ligand-1(PSGL-1), and regulated by hemodynamics. Poor data were reported in understanding P-selectin-mediated rolling. With the flow chamber technique, we herein observed HL-60 cell rolling on P-selectin with or without 3% Ficoll at various wall shear stresses from 0.05 to 0.4 dyn/cm2. The results demonstrated that force rather than transport regulated the rolling, similar to rolling on L- and E-selectin. The rolling was accelerated quickly by an increasing force below the optimal shear threshold of 0.15 dyn/cm2 first and then followed by a slowly decelerating phase starting at the optimum, showing a catch-slip transition and serving as a mechanism for the rolling. The catch-slip transition was completely reflected to the tether lifetime and other rolling parameters, such as the mean and fractional stop time. The narrow catch bond regime stabilized the rolling quickly, through steeply increasing fractional stop time to a plateau of about 0.85. Data presented here suggest that the low shear stress threshold serves as a mechanism for most cell rolling events through P-selectin.
The velocity profile around cells in a flow chamber coated with the immobilized protein and the endothelial cells is studied using the micro particle image velocimetry(PIV). The main purpose is to study the effect of the endothelial cells on the local hydrodynamic environment and the local shear rates above a single polymorphonuclear neutrophil(PMN) and a melanoma cell when they adhere to different immobilized protein substrates. Micro-PIV images are taken in the top-view and the side-view under 10 X and 40 X objective lens and the ensemble correlation method is used to analyze the data. The results show that the endothelial monolayer has changed the patterns of the flow velocity profile of the side-view flow on the chamber bottom, and also increased the wall shear rates. The melanoma cells adhered on the immobilized fibrin disturb the local flow more than those adhered on the immobilized fibrinogen, but one sees no significant difference between the local shear rates above the PMNs adhered on the immobilized fibrinogen and those above the PMNs adhered on the immobilized fibrin.
