Coimmobilized native macromolecular heparin proteoglycans strongly inhibitplatelet-collagen interactions in flowing blood

We coimmobilized mast cell-derived heparin proteoglycans (HEP-PGs) of very high molecular weight (750 kDa) or unfractionated heparin (UFH) on coversli ps together with collagen without altering the amount of immobilized collag en. Subsequently, platelet-collagen interactions were studied under both fl owing and static conditions in D-phenylalanyl-L-prolyl-L-arginine chloromet hyl ketone-anticoagulated blood and platelet-rich plasma (PRP), respectivel y. At a high shear rate (1600 1/s), the mean platelet deposition (PD) on co llagen monomers was 7.5+/-6.1x10(6)/cm(2) (n=5). When the monomers were coi mmobilized with UFH, PD was inhibited by 73% (2.0+/-1.2x10(6)/cm(2)), where as HEP-PG completely blocked it (0.42+/-0.38x10(6)/cm(2); P<0.05). Also, wh en collagen fibrils were used for coating, HEP-PG significantly inhibited P D. At a low shear rate (200 1/s) and under static conditions in PRP, the in hibitory effect of HEP-PG on PD was less marked. Inhibition of glycoprotein IIb/IIIa did not affect PD on coimmobilized HEP-PG in contrast to coimmobi lized UFH or collagen alone. As a sign of inactivation, platelets adhering to the HEP-PG surface released considerably less <beta>-thromboglobulin tha n did those adhering to pure collagen. In summary, immobilized HEP-PG stron gly inhibited PD on collagen by attenuating adhesion-induced platelet activ ation. The stronger effect on collagen monomers suggests the inhibition of glycoprotein Ia/IIa-mediated activation.