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From the Research Forum, Ullevaal University Hospital (I.A.H., T.L.), and the Department of Plastic Surgery, Rikshospitalet National Hospital (H.E.R.), Oslo, Norway.
Correspondence to Inger Anne Hagberg, Research Forum, Ullevaal University Hospital, 0407 Oslo, Norway.
Abstract We investigated the combined effect of wall shear
rate and immobilized collagen on platelet activation in
flowing nonanticoagulated human blood. By combining an ex vivo model of
thrombogenesis with flow cytometry, we showed that activated
platelets can be detected in the bloodstream passing growing
thrombi at a wall shear rate characteristic of moderately stenosed
arteries (2600 s-1). The activation of the
circulating platelets was clearly correlated with thrombus growth.
Different antibodies against platelet activation-dependent surface
markers had distinct sensitivity to the thrombotic process.
-Granule
release detected by surface expression of CD62P seemed to be the most
sensitive marker, as judged by both mean fluorescence intensity
and fraction of platelets activated. The conformational
change in glycoprotein IIbIIIa, as detected by PAC-1,
also seemed to be a sensitive marker and preceded binding of fibrinogen
to activated glycoprotein IIbIIIa, as detected by
anti-fibrinogen. Large thrombi also elicited lysosome
exocytosis, detected by surface expression of CD63. Finally, we
observed a small decrease of glycoprotein IbIX
expression, as detected by anti-CD42a. Thus, our study provides further
information on the dynamics of platelet activation in relation to
thrombus growth at arterial shear conditions in flowing
nonanticoagulated human blood.
Key Words: perfusion platelet activation flow cytometry arterial thrombogenesis collagen
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