title: Aspects of hydrodynamic shear regulating shear-induced platelet activation and self-association of von Willebrand factor in suspension.
abstract: The binding of plasma von Willebrand factor (VWF) to platelet receptor GpIb under high hydrodynamic shear leads to platelet activation and subsequent shear-induced platelet aggregation (SIPA). We quantitatively examined the aspects of fluid flow that regulate platelet activation by subjecting human blood and isolated platelets to well-defined shear conditions in a cone-plate viscometer. We made the following observations. First, Annexin V binding to phosphatidyl serine expressed on activated cells was detectable within 10 seconds of shear application. Second, fluid shear stress rather than shear rate controls platelet activation, and a threshold shear stress of approximately 80 dyn/cm(2) is necessary to induce significant activation. Under these conditions, individual domains of soluble VWF and platelet GpIb are subjected to similar magnitudes of fluid forces on the order of 0.1 pN, whereas GpIb with bound VWF is subjected to 1 pN. Third, cell-cell collisions and time-varying stresses are not essential for platelet activation. Fourth, the mechanism of platelet activation can be resolved in 2 steps based on the contribution of VWF and fluid forces. Fluid shear and VWF are required during the first step, when GpIb-VWF binding likely occurs. Subsequently, high shear forces alone in the absence of VWF in suspension can induce platelet activation. In other experiments, purified VWF was subjected to shear in the viscometer, and VWF morphology was assessed using light scattering. These studies demonstrate, for the first time, the ability of hydrodynamic forces to induce VWF aggregation in suspension. This VWF self-association may be an additional feature involved in controlling cell adhesion rates in circulation.
authors.affiliation: Bioengineering Laboratory, Department of Chemical Engineering, State University of New York at Buffalo, NY 14260, USA.
authors.forename: Harish
authors.initials: H
authors.lastname: Shankaran
authors.forename: Paschalis
authors.initials: P
authors.lastname: Alexandridis
authors.forename: Sriram
authors.initials: S
authors.lastname: Neelamegham
mesh_terms: D017304:Annexin A5 / Q000378:metabolism; D019281:Dimerization; D006801:Humans; D007700:Kinetics; D015539:Platelet Activation; D011485:Protein Binding; D012212:Rheology; D013314:Stress, Mechanical; D013535:Suspensions; D014783:Viscosity; D014841:von Willebrand Factor / Q000378:metabolism*
chemical_list: D017304:Annexin A5; D013535:Suspensions; D014841:von Willebrand Factor