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Thromb Haemost 1998; 79(02): 296-301
DOI: 10.1055/s-0037-1614981
Letters to the Editor
Schattauer GmbH

Initiation and Propagation of Blood Coagulation at Artificial Surfaces Studied in a Capillary Flow Reactor[*]

Ron Blezer
1   From the Dept. of Biochemistry
,
George M. Willems
2   From the Cardiovascular Research Institute Maastricht, Maastricht University
,
Patrick T. Cahalan
3   From the Dept. of Biomaterials Technology, Medtronic Bakken Research Center, Maastricht, The Netherlands
,
Theo Lindhout
1   From the Dept. of Biochemistry
2   From the Cardiovascular Research Institute Maastricht, Maastricht University
› Author Affiliations
Further Information

Publication History

Received 26 May 1997

Accepted after revision 16 September 1997

Publication Date:
08 December 2017 (online)

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Summary

We have made use of a novel flow reactor to study the initiation and propagation of the ex vivo blood coagulation processes at artificial surfaces. The flow reactor consisted of a primary glass or polymer capillary that is connected to a secondary glass capillary, which inner wall was coated with a phospholipid bilayer of 25 mol% dioleoylphosphatidylserine/75 mol% dioleoylphosphatidylcholine (DOPS/DOPC). Citrated platelet free plasma and a CaCl2 solution were delivered by syringe pumps and mixed just before the entrance of the flow reactor. The outflowing plasma was assayed for factor XIa, factor IXa, factor Xa and thrombin activity. Perfusion of recalcified plasma through a bare glass capillary resulted in a transient generation of fluid phase factor XIa. In contrast, factor IXa production increased slowly to attain a stable steady-state level. We established that surface-bound factor XIa was responsible for a continuous production of factor IXa. Factor IXa-induced generation of factor Xa and thrombin was only observed when contact activated plasma was subsequently perfused through a DOPS/DOPC-coated capillary, showing that propagation of the factor IXa trigger requires a procoagulant, phosphatidylserine-containing, phospholipid membrane. The negatively charged inner surface of a heparin-coated polyurethane capillary, generated like the glass capillary significant amounts of factor XIa and factor IXa when perfused with recalcified plasma. No differences were found between unfractionated heparin and heparin devoid of anticoagulant activity. Thus, it is concluded that contact activation and factor IXa generation in flowing plasma is not inhibited by immobilised anticoagulant active heparin. Consequently, factor IXa-dependent thrombin generation at a downstream located phospholipid membrane was similar, regardless the specific anticoagulant activity of immobilised heparin.

* Supported by grant 902-26-192 from the Dutch Organization for Scientific Research (NWO)


 

  • References

  • 1 Nemerson Y. The tissue factor pathway of blood coagulation. Semin Hematol 1992; 29: 170-88.
    PubMedSearch in Google Scholar
  • 2 Forbes CD, Courtney JM. Thrombosis and artificial surfaces. In: Haemostasis and Thrombosis. Bloom AL, and Thomas ED. eds. Edinburgh: Churchill Livingstone; 1994. pp 1301-24.
    Search in Google Scholar
  • 3 Mann KG, Krishnaswamy S, Lawson JH. Surface-dependent hemostasis. Sem Hematol 1992; 29: 213-26.
    PubMedSearch in Google Scholar
  • 4 Hemker HC, BÈguin S. 1994. Thrombin generation an essential step in haemostasis and thrombosis. In: Haemostasis and Thrombosis. Bloom AL, and Thomas ED. eds. Edinburgh: Churchill Livingstone; 1994. pp 447-90.
    Search in Google Scholar
  • 5 Zwaal RFA, Comfurius P, Bevers EM. Platelet procoagulant activity and microvesicle formation – its putative role in hemostasis and thrombosis. Biochim Biophys Acta 1992; 1180: 1-8.
    CrossrefPubMedSearch in Google Scholar
  • 6 Gemmell CH, Broze GJ, Turitto VT, Nemerson Y. Utilization of a continuous flow reactor to study the lipoprotein-associated coagulation inhibitor (LACI) that inhibits tissue factor. Blood 1990; 76: 2266-71.
    PubMedSearch in Google Scholar
  • 7 Billy D, Speijer H, Willems GM, Hemker HC, Lindhout T. Prothrombin activation by prothrombinase in a tubular flow reactor. J Biol Chem 1995; 270: 1029-34.
    CrossrefPubMedSearch in Google Scholar
  • 8 Speijer H, Billy D, Willems GM, Hemker HC, Lindhout T. Inhibition of prothrombinase by antithrombin-heparin at a macroscopic surface. Thromb Haemost 1995; 73: 648-53.
    Thieme ConnectPubMedSearch in Google Scholar
  • 9 Béguin S, Lindhout T, Hemker HC. The mode of action of heparin in plasma. Thromb Haemost 1988; 60: 457-62.
    Thieme ConnectPubMedSearch in Google Scholar
  • 10 Lindhout T, Blezer R, Willems GM, Fouache B, Verhoeven M, Cahalan L, Cahalan PT. Antithrombin activity of surface-bound heparin studied under flow conditions. J Biomed Mater Res 1995; 29: 1255-66.
    CrossrefPubMedSearch in Google Scholar
  • 11 Wuillemin WA, Minnema M, Meijers JC, Roem D, Eerenberg AJ, Nuijens JH, ten Cate H, Hack CE. Inactivation of factor XIa in human plasma assessed by measuring factor XIa-protease inhibitor complexes: major role for C1-inhibitor. Blood 1995; 85: 1517-26.
    PubMedSearch in Google Scholar
  • 12 Pieters J, Lindhout T. The limited importance of factor Xa inhibition to the anticoagulant property of heparin in thromboplastin-activated plasma. Blood 1988; 72: 2048-52.
    PubMedSearch in Google Scholar
  • 13 Wiggins RC, Bouma BN, Cochrane CG, Griffin JH. Role of high molecular weight kininogen in surface-binding and activation of coagulation factor XI and prekallikrein. Proc Natl Acad Sci USA 1977; 74: 4636-40.
    CrossrefPubMedSearch in Google Scholar
  • 14 van der Graaf F, Greengard JS, Bouma BN, Kerbiriou DM, Griffin JH. Isolation and functional characterization of the active light chain of activated human blood coagulation factor XI. J Biol Chem 1983; 258: 9669-75.
    PubMedSearch in Google Scholar
  • 15 Walsh PN, Sinha D, Koshy A, Seaman FS, Bradford H. Functional characterization of platelet-bound factor XIa: retention of factor XIa activity on the platelet surface. Blood 1986; 68: 225-30.
    PubMedSearch in Google Scholar
  • 16 Pieters J, Lindhout T, Hemker HC. In situ generated thrombin is the only enzyme that effectively activates factor VIII and Factor V in thromboplastin-activated plasma. Blood 1989; 74: 1021-4.
    PubMedSearch in Google Scholar
  • 17 Willems GM, Lindhout T, Hermens WT, Hemker HC. Simulation model for thrombin generation in plasma. Haemostasis 1991; 21: 197-207.
    PubMedSearch in Google Scholar
  • 18 Blezer R, Fouache B, Willems GM, Lindhout T. Activation of blood coagulation at heparin-coated surfaces. J Biomed Mater Res 1997; 37: 108-13.
    CrossrefPubMedSearch in Google Scholar
  • 19 Sanchez J, Elgue G, Riesenfeld J, Olsson P. Control of contact activation on end-point immobilized heparin: the role of antithrombin and the specific antithrombin-binding sequence. J Biomed Mater Res 1995; 29: 655-61.
    CrossrefPubMedSearch in Google Scholar
  • 20 Griffin JH, Cochrane CG. Mechanisms for the involvement of high molecular weight kininogen in surface-dependent reactions of Hageman factor. Proc Natl Acad Sci USA 1976; 73: 2554-8.
    CrossrefPubMedSearch in Google Scholar
  • 21 Dors DM, Nuijens JH, Huijbregts CCM, Hack CE. A Novel sensitive assay for functional factor-XII based on the generation of kallikrein-C1-inhibitor complexes in factor-XII-deficient plasma by glass-bound factor-XII. Thromb Haemost 1992; 67: 644-8.
    Thieme ConnectPubMedSearch in Google Scholar
  • 22 Naito K, Fujikawa K. Activation of human blood coagulation factor XI independent of factor XII. Factor XI is activated by thrombin and factor XIa in the presence of negatively charged surfaces. J Biol Chem 1991; 266: 7353-8.
    PubMedSearch in Google Scholar
  • 23 Broze GJ, Gailani D. The role of Factor-XI in coagulation. Thromb Haemost 1993; 70: 72-4.
    Thieme ConnectPubMedSearch in Google Scholar
  • 24 Brunnee T, La Porta C, Reddigari SR, Salerno VM, Kaplan AP, Silverberg M. Activation of factor XI in plasma is dependent on factor XII. Blood 1993; 81: 580-6.
    PubMedSearch in Google Scholar