Fibrinogen Adsorption, Platelet Adhesion and Thrombin Generation at Heparinized Surfaces Exposed to Flowing Blood

Jeffrey F. W. Keuren; Simone J. H. Wielders; George M. Willems; Marco Morra; Theo Lindhout

doi:10.1055/s-0037-1613074

Thrombosis and Haemostasis, Inhaltsverzeichnis

Thromb Haemost 2002; 87(04): 742-747
DOI: 10.1055/s-0037-1613074

Review Article

Schattauer GmbH

Fibrinogen Adsorption, Platelet Adhesion and Thrombin Generation at Heparinized Surfaces Exposed to Flowing Blood

Jeffrey F. W. Keuren^*

¹Department of Biochemistry and Maastricht University, Maastricht, The Netherlands

²Cardiovascular Research Institute, Maastricht University, Maastricht, The Netherlands

,

Simone J. H. Wielders^*

¹Department of Biochemistry and Maastricht University, Maastricht, The Netherlands

,

George M. Willems

²Cardiovascular Research Institute, Maastricht University, Maastricht, The Netherlands

,

Marco Morra

³Nobil Bio Ricerche, Villafranca d’Asti, Italy

,

Theo Lindhout

¹Department of Biochemistry and Maastricht University, Maastricht, The Netherlands

²Cardiovascular Research Institute, Maastricht University, Maastricht, The Netherlands

› Institutsangaben

Abstract

Summary

Thrombus formation at an artificial surface in contact with blood is a complex process that encompasses accretion of platelets from flowing blood and fibrin deposition. Platelet adhesion and fibrin formation are intimately intertwined reactions that are triggered by different sets of surface adsorbed plasma proteins. To dissect the contribution of protein adsorption and platelet adhesion to thrombin formation, a coherent study was performed with non-coated (NC) and heparin-coated (HC) surfaces. Thrombin production in whole blood, platelet adhesion and protein adsorption were studied using an amidolytic thrombin assay, a dynamic platelet adhesion assay and ellipsometry, respectively. Thrombin generation in flowing whole blood exposed to HC surfaces was greatly diminished when compared with NC surfaces. However, separate platelet adhesion and protein adsorption studies with anticoagulated whole blood revealed that platelets do not adhere because fibrinogen is not available in the protein layer that was deposited during the perfusion. These findings indicate that the in vitro thrombogenicity of a material cannot be predicted from platelet adhesion and protein adsorption data when these measurements are performed with anticoagulated blood or platelet rich plasma. Preincubation of NC and HC surfaces with fibrinogen or 2000-fold diluted plasma resulted in similar amounts of surface-bound fibrinogen and mediated massive platelet adhesion from flowing whole blood. These results indicate that a) platelet adhesion correlates with the availability of surface-bound fibrinogen and b) NC and HC surfaces are indistinguishable with respect to protein (fibrinogen) adsorption and platelet adhesion. It is apparent that the heparinized surface used in our studies exerts its anti-thrombogenic properties by neutralizing locally formed thrombin and not by reducing fibrinogen-dependent platelet adhesion.

Keywords

Heparin coating - artificial surface - fibrinogen - platelets - thrombin

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Referenzen

References
1 Lamba NMK, Cooper SL. Interaction of blood with artificial surfaces. In: Hemostasis and thrombosis. Basic principles and clinical practice. Colman RW, Hirsh J, Marder VJ, Clowes AW, George JN. eds. Philadelphia: Lippincott Williams & Wilkins; 2001: 661-72.
2 Hanson SR, Harker LA, Ratner BD, Hoffman AS. In vivo evaluation of artificial surfaces using a nonhuman primate model of arterial thrombosis. J Lab Clin Med 1980; 95: 289-304.
3 Plow EF, McEver RP, Coller BS, Woods VL, Marguerie GA, Ginsberg MH. Related binding mechanisms for fibrinogen, fibronectin, von Willebrand factor, and thrombospondin on thrombin-stimulated platelets. Blood 1985; 66: 724-7.
4 Goodman SL, Cooper SL, Albrecht RM. Integrin receptors and platelet adhesion to synthetic surfaces. J Biomed Mater Res 1993; 27: 683-95.
5 Wagner WR, Hubbell JA. Local thrombin synthesis and fibrin formation in an in vitro thrombosis model results in platelet recruitment and thrombus stabilization on collagen in heparinized blood. J Lab Clin Med 1990; 116: 636-50.
6 Grunkemeier JM, Tsai WB, Horbett TA. Hemocompatibility of treated polystyrene substrates: contact activation, platelet adhesion, and procoagulant activity of adherent platelets. J Biomed Mater Res 1998; 41: 657-70.
7 Colman RW. Contact activation pathway: inflammatory, fybrinolytic, anticoagulant, antiadhesive and antiangiogenic activities. In: Hemostasis and thrombosis. Basic principles and clinical practice. Colman RW, Hirsh J, Marder VJ, Clowes AW, George JN. eds. Philadelphia: Lippincott Williams & Wilkins; 2001: 103-21.
8 Andrade JD. Principles of protein adsorption. In: Surface and Interfacial Aspects of Biomedical Polymers. Andrade J. ed. New York: Plenum Publ; 1985: 1-80.
9 Horbett TA, Brash JL. Proteins at interfaces: current issues and future aspects. In: Proteins at interfaces. Physiochemical and biochemical studies. Horbett TA, Brash JL. eds. Washington, DC: American Chemical Society; 1987: 1-33.
10 Green RJ, Davies J, Davies MC, Roberts CJ, Tendler SJ. Surface plasmon resonance for real time in situ analysis of protein adsorption to polymer surfaces. Biomaterials 1997; 18: 405-13.
11 Jen CJ, Lin JS. Direct observation of platelet adhesion to fibrinogenand fibrin-coated surfaces. Am J Physiol 1991; 261: H1457-63.
12 Endenburg SC, Lindeboom LBlokzijl, Zwaginga JJ, Sixma JJ, de Groot PG. Plasma fibrinogen inhibits platelets adhesion in flowing blood to immobilized fibrinogen. Arterioscler Thromb Vasc Biol 1996; 16: 633-8.
13 Feuerstein IA, McClung WG, Horbett TA. Platelet adherence and detachment with adsorbed fibrinogen: a flow study with a series of hydroxyethyl methacrylate-ethyl methacrylate copolymers using video microscopy. J Biomed Mater Res 1992; 26: 221-37.
14 Lee JH, Ju YM, Kim DM. Platelet adhesion onto segmented polyurethane film surfaces modified by addition and crosslinking of PEO-containing block copolymers. Biomaterials 2000; 21: 683-91.
15 Grunkemeier JM, Tsai WB, McFarland CD, Horbett TA. The effect of adsorbed fibrinogen, fibronectin, von Willebrand factor and vitronectin on the procoagulant state of adherent platelets. Biomaterials 2000; 21: 2243-52.
16 Vroman L, Adams AL, Fischer GC, Munoz PC. Interaction of high molecular weight kininogen, factor XII, and fibrinogen in plasma at interfaces. Blood 1980; 55: 156-9.
17 Cuypers PA, Willems GM, Hemker HC, Hermens WT. Adsorption kinetics of protein mixtures. A tentative explanation of the Vroman effect. Ann N Y Acad Sci 1987; 516: 244-52.
18 Chinn JA, Posso SE, Horbett TA, Ratner BD. Postadsorptive transitions in fibrinogen adsorbed to polyurethanes: changes in antibody binding and sodium dodecyl sulfate elutability. J Biomed Mater Res 1992; 26: 757-78.
19 Slack SM, Horbett TA. The Vroman effect: a critical review. In: Proteins at interfaces. II: fundamentals and application. Brash TA, Brash JL. eds. Washington, DC: American Chemical Society; 1995: 112-8.
20 Balasubramanian V, Grusin NK, Bucher RW, Turitto VT, Slack SM. Residence-time dependent changes in fibrinogen adsorbed to polymeric biomaterials. J Biomed Mater Res 1999; 44: 253-60.
21 Billy D, Briede J, Heemskerk JWM, Hemker HC, Lindhout T. Prothrombin conversion under flow conditions by prothrombinase assembled on adherent platelets. Blood Coagul Fibrinolysis 1997; 08: 168-74.
22 Schoen P, Lindhout T, Willems G, Hemker HC. Continuous flow and the prothrombinase-catalyzed activation of prothrombin. Thromb Haemost 1990; 64: 542-7.
23 Heemskerk JW, Vuist WM, Feijge MA, Reutelingsperger CP, Lindhout T. Collagen but not fibrinogen surfaces induce bleb formation, exposure of phosphatidylserine, and procoagulant activity of adherent platelets: evidence for regulation by protein tyrosine kinase-dependent Ca2+ responses. Blood 1997; 90: 2615-25.
24 Cuypers PA, Corsel JW, Janssen MP, Kop JM, Hermens WT, Hemker HC. The adsorption of prothrombin to phosphatidylserine multilayers quantitated by ellipsometry. J Biol Chem 1983; 258: 2426-31.
25 Blezer R, Cahalan L, Cahalan PT, Lindhout T. Heparin coating of tantalum coronary stents reduces surface thrombin generation but not factor IXa generation. Blood Coagul Fibrinolysis 1998; 09: 435-40.
26 Blezer R, Willems GM, Cahalan PT, Lindhout T. Initiation and propagation of blood coagulation at artificial surfaces studied in a capillary flow reactor. Thromb Haemost 1998; 79: 296-301.
27 Horbett TA. Principles underlying the role of adsorbed plasma proteins in blood interactions with foreign materials. Cardiovascular Pathology 1993; 02: 137S-48S.
28 Adams AL, Fischer GC, Munoz PC, Vroman L. Convex-lens-on-slide: a simple system for the study of human plasma and blood in narrow spaces. J Biomed Mater Res 1984; 18: 643-54.
29 Brash JL, ten Hove P. Effect of plasma dilution on adsorption of fibrinogen to solid surfaces. Thromb Haemost 1984; 51: 326-30.
30 Horbett TA. Mass action effects on competitive adsorption of fibrinogen from hemoglobin solutions and from plasma. Thromb Haemost 1984; 51: 174-81.