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. 1985 Feb;75(2):531–540. doi: 10.1172/JCI111729

Role of factor VIII-von Willebrand factor and fibronectin in the interaction of platelets in flowing blood with monomeric and fibrillar human collagen types I and III.

W P Houdijk, K S Sakariassen, P F Nievelstein, J J Sixma
PMCID: PMC423528  PMID: 3919060

Abstract

Platelet adhesion to monomeric collagen types I and III, which were purified from human umbilical arteries, was studied in a perfusion chamber under well defined flow conditions. For this purpose, glass coverslips were coated with 20-30 micrograms/cm2 of collagen types I and III by spraying a solution of these collagens with a retouching air brush. Platelet deposition increased with the time of perfusion. Adhesion to both collagen types was similar in the first 3 min, but increased platelet deposition occurred on collagen type III after 3 min due to thrombus formation. Adhesion at a shear rate of 800 s-1 was strongly impaired with plasma of a patient with von Willebrand's disease or with fibronectin-free plasma. Addition of purified fibronectin to fibronectin-free plasma restored adhesion to the level obtained with normal plasma. Platelet deposition in normal plasma increased with increasing shear rates. Platelet deposition in VWD-plasma was normal at 490 s-1, but there was no increase at higher shear rates. Platelet deposition in fibronectin-free plasma was diminished at all shear rates studied from 490 to 1,300 s-1. Perfusion with a human albumin solution (HAS) to which purified Factor VIII-von Willebrand factor complex (FVIII-VWF) and fibronectin had been added gave similar platelet deposition as with normal plasma. Preincubation of collagen with FVIII-VWF and perfusion with HAS containing fibronectin, or, conversely, preincubation with fibronectin and perfusion with HAS containing FVIII-VWF, also resulted in adhesion similar to that observed in normal plasma. Similar adhesion was also observed after preincubation with both FVIII-VWF and fibronectin and subsequent perfusion with HAS alone. Sequential preincubations with first FVIII-VWF and then fibronectin, or with first fibronectin and then FVIII-VWF followed by perfusion with HAS, also gave a similar adhesion as observed with normal plasma. These data indicate that platelet adhesion to monomeric collagen types I and III is dependent on both FVIII-VWF and fibronectin. FVIII-VWF is only required at relatively high shear rates; fibronectin also at relatively low shear rates. Their complementary role in platelet adhesion suggests separate binding sites for FVIII-VWF and fibronectin on collagen. Platelet deposition on performed fibrils of collagen types I and III was also studied. Initial adhesion expressed as percentage surface coverage was similar to that found with monomeric collagen, but thrombus formation was much enhanced. Adhesion on fibrillar collagen at 800 s(-1) was impaired in VWD-plasma and fibronectin-free plasma, and was restored by addition of purified fibronectin to fibronectin-free plasma. When perfusions were performed with HAS, only addition of FVIII-VWF was required for optimal adhesion to fibrillar collagen; addition of fibronectin had no effect. These data are in contrast to the studies with monomeric collagens described above, in which the addition of both FVIII-VWF and fibronectin was required. These data are also in contrast to the observation that in plasma both FVIII-VWF and fibronectin are required for optimal adhesion to fibrillar collagen.

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Selected References

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  1. Allain J. P., Cooper H. A., Wagner R. H., Brinkhous K. M. Platelets fixed with paraformaldehyde: a new reagent for assay of von Willebrand factor and platelet aggregating factor. J Lab Clin Med. 1975 Feb;85(2):318–328. [PubMed] [Google Scholar]
  2. Amrani D. L., Homandberg G. A., Tooney N. M., Wolfenstein-Todel C., Mosesson M. W. Separation and analysis of the major forms of plasma fibronectin. Biochim Biophys Acta. 1983 Oct 28;748(2):308–320. doi: 10.1016/0167-4838(83)90308-4. [DOI] [PubMed] [Google Scholar]
  3. Aplin J. D., Hughes R. C. Protein-derivatised glass coverslips for the study of cell-to substratum adhesion. Anal Biochem. 1981 May 1;113(1):144–148. doi: 10.1016/0003-2697(81)90057-9. [DOI] [PubMed] [Google Scholar]
  4. Balian G., Click E. M., Bornstein P. Location of a collagen-binding domain in fibronectin. J Biol Chem. 1980 Apr 25;255(8):3234–3236. [PubMed] [Google Scholar]
  5. Baumgartner H. R. Platelet interaction with collagen fibrils in flowing blood. I. Reaction of human platelets with alpha chymotrypsin-digested subendothelium. Thromb Haemost. 1977 Feb 28;37(1):1–16. [PubMed] [Google Scholar]
  6. Baumgartner H. R., Tschopp T. B., Weiss H. J. Platelet interaction with collagen fibrils in flowing blood. II. Impaired adhesion-aggregation in bleeding disorders. A comparison with subendothelium. Thromb Haemost. 1977 Feb 28;37(1):17–28. [PubMed] [Google Scholar]
  7. Bensusan H. B., Koh T. L., Henry K. G., Murray B. A., Culp L. A. Evidence that fibronectin is the collagen receptor on platelet membranes. Proc Natl Acad Sci U S A. 1978 Dec;75(12):5864–5868. doi: 10.1073/pnas.75.12.5864. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Bolhuis P. A., Sakariassen K. S., Sander H. J., Bouma B. N., Sixma J. J. Binding of factor VIII-von Willebrand factor to human arterial subendothelium precedes increased platelet adhesion and enhances platelet spreading. J Lab Clin Med. 1981 Apr;97(4):568–576. [PubMed] [Google Scholar]
  9. Booyse F. M., Feder S., Quarfoot A. J. Culture-produced subendothelium. II. Effect of plasma, F.VIIIR:WF and fibronectin on interaction of normal platelets with normal and von Willebrand porcine aortic subendothelium. Thromb Res. 1982 Nov 1;28(3):299–311. doi: 10.1016/0049-3848(82)90113-x. [DOI] [PubMed] [Google Scholar]
  10. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
  11. Bruns R. R., Gross J. High-resolution analysis of the modified quarter-stagger model of the collagen fibril. Biopolymers. 1974 May;13(5):931–941. doi: 10.1002/bip.1974.360130509. [DOI] [PubMed] [Google Scholar]
  12. Cejka J. Enzyme immunoassay for factor VIII-related antigen. Clin Chem. 1982 Jun;28(6):1356–1358. [PubMed] [Google Scholar]
  13. ChandraRajan J. Separation of type III collagen from type I collagen and pepsin by differential denaturation and renaturation. Biochem Biophys Res Commun. 1978 Jul 14;83(1):180–186. doi: 10.1016/0006-291x(78)90414-x. [DOI] [PubMed] [Google Scholar]
  14. Chung E., Miller E. J. Collagen polymorphism: characterization of molecules with the chain composition (alpha 1 (3)03 in human tissues. Science. 1974 Mar;183(130):1200–1201. doi: 10.1126/science.183.4130.1200. [DOI] [PubMed] [Google Scholar]
  15. Dessau W., Adelmann B. C., Timpl R. Identification of the sites in collagen alpha-chains that bind serum anti-gelatin factor (cold-insoluble globulin). Biochem J. 1978 Jan 1;169(1):55–59. doi: 10.1042/bj1690055. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Engvall E., Ruoslahti E., Miller E. J. Affinity of fibronectin to collagens of different genetic types and to fibrinogen. J Exp Med. 1978 Jun 1;147(6):1584–1595. doi: 10.1084/jem.147.6.1584. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Grinnell F., Feld M., Snell W. The influence of cold insoluble globulin on platelet morphological response to substrata. Cell Biol Int Rep. 1979 Oct;3(7):585–592. doi: 10.1016/0309-1651(79)90056-0. [DOI] [PubMed] [Google Scholar]
  18. Heyns A. D., Badenhorst P. N., Pieters H., Lötter M. G., Minnaar P. C., Duyvené de Wit L. J., Van Reenen O. R., Retief F. P. Preparation of a viable population of indium-111-labelled human blood platelets. Thromb Haemost. 1980 Feb 29;42(5):1473–1482. [PubMed] [Google Scholar]
  19. Hovig T., Stormorken H. Ultrastructural studies on the platelet plug formation in bleeding time wounds from normal individuals and patients with von Willebrand's disease. Acta Pathol Microbiol Scand Suppl. 1974;Suppl 248:105–122. [PubMed] [Google Scholar]
  20. Hynes R. O., Ali I. U., Destree A. T., Mautner V., Perkins M. E., Senger D. R., Wagner D. D., Smith K. K. A large glycoprotein lost from the surfaces of transformed cells. Ann N Y Acad Sci. 1978 Jun 20;312:317–342. doi: 10.1111/j.1749-6632.1978.tb16811.x. [DOI] [PubMed] [Google Scholar]
  21. Klebe R. J., Bentley K. L., Sasser P. J., Schoen R. C. Elution of fibronectin from collagen with chaotrophic agents. Exp Cell Res. 1980 Nov;130(1):111–117. doi: 10.1016/0014-4827(80)90047-6. [DOI] [PubMed] [Google Scholar]
  22. Kleinman H. K., Klebe R. J., Martin G. R. Role of collagenous matrices in the adhesion and growth of cells. J Cell Biol. 1981 Mar;88(3):473–485. doi: 10.1083/jcb.88.3.473. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Kleinman H. K., McGoodwin E. B., Martin G. R., Klebe R. J., Fietzek P. P., Woolley D. E. Localization of the binding site for cell attachment in the alpha1(I) chain of collagen. J Biol Chem. 1978 Aug 25;253(16):5642–5646. [PubMed] [Google Scholar]
  24. Koteliansky V. E., Leytin V. L., Sviridov D. D., Repin V. S., Smirnov V. N. Human plasma fibronectin promotes the adhesion and spreading of platelets on surfaces coated with fibrillar collagen. FEBS Lett. 1981 Jan 12;123(1):59–62. doi: 10.1016/0014-5793(81)80019-1. [DOI] [PubMed] [Google Scholar]
  25. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  26. Lahav J., Schwartz M. A., Hynes R. O. Analysis of platelet adhesion with a radioactive chemical crosslinking reagent: interaction of thrombospondin with fibronectin and collagen. Cell. 1982 Nov;31(1):253–262. doi: 10.1016/0092-8674(82)90425-1. [DOI] [PubMed] [Google Scholar]
  27. Laurell C. B. Electroimmuno assay. Scand J Clin Lab Invest Suppl. 1972;124:21–37. doi: 10.3109/00365517209102748. [DOI] [PubMed] [Google Scholar]
  28. Legrand Y. J., Rodriguez-Zeballos A., Kartalis G., Fauvel F., Caen J. P. Adsorption of factor VIII antigen-activity complex by collagen. Thromb Res. 1978 Nov;13(5):909–911. doi: 10.1016/0049-3848(78)90196-2. [DOI] [PubMed] [Google Scholar]
  29. Leytin V. L., Gorbunova N. A., Misselwitz F., Novikov I. D., Podrez E. A., Plyusch O. P., Likhachova E. A., Repin V. S., Smirnov V. N. Step-by-step analysis of adhesion of human platelets to a collagen-coated surface defect in initial attachment and spreading of platelets in von Willebrand's disease. Thromb Res. 1984 Apr 1;34(1):51–63. doi: 10.1016/0049-3848(84)90105-1. [DOI] [PubMed] [Google Scholar]
  30. Macfarlane D. E., Stibbe J., Kirby E. P., Zucker M. B., Grant R. A., McPherson J. Letter: A method for assaying von Willebrand factor (ristocetin cofactor). Thromb Diath Haemorrh. 1975 Sep 30;34(1):306–308. [PubMed] [Google Scholar]
  31. Mosher D. F., Johnson R. B. In vitro formation of disulfide-bonded fibronectin multimers. J Biol Chem. 1983 May 25;258(10):6595–6601. [PubMed] [Google Scholar]
  32. Muggli R., Baumgartner H. R., Tschopp T. B., Keller H. Automated microdensitometry and protein assays as a measure for platelet adhesion and aggregation on collagen-coated slides under controlled flow conditions. J Lab Clin Med. 1980 Feb;95(2):195–207. [PubMed] [Google Scholar]
  33. Over J., Sixma J. J., Bruïne M. H., Trieschnigg M. C., Vlooswijk R. A., Beeser-Visser N. H., Bouma B. N. Survival of 125iodine-labeled Factor VIII in normals and patients with classic hemophilia. Observations on the heterogeneity of human Factor VIII. J Clin Invest. 1978 Aug;62(2):223–234. doi: 10.1172/JCI109120. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Plow E. F., Ginsberg M. H. Specific and saturable binding of plasma fibronectin to thrombin-stimulated human platelets. J Biol Chem. 1981 Sep 25;256(18):9477–9482. [PubMed] [Google Scholar]
  35. Sakariassen K. S., Aarts P. A., de Groot P. G., Houdijk W. P., Sixma J. J. A perfusion chamber developed to investigate platelet interaction in flowing blood with human vessel wall cells, their extracellular matrix, and purified components. J Lab Clin Med. 1983 Oct;102(4):522–535. [PubMed] [Google Scholar]
  36. Sakariassen K. S., Bolhuis P. A., Sixma J. J. Human blood platelet adhesion to artery subendothelium is mediated by factor VIII-Von Willebrand factor bound to the subendothelium. Nature. 1979 Jun 14;279(5714):636–638. doi: 10.1038/279636a0. [DOI] [PubMed] [Google Scholar]
  37. Sakariassen K. S., Bolhuis P. A., Sixma J. J. Platelet adherence to subendothelium of human arteries in pulsatile and steady flow. 1980 Aug 15-Sep 1Thromb Res. 19(4-5):547–559. doi: 10.1016/0049-3848(80)90027-4. [DOI] [PubMed] [Google Scholar]
  38. Santoro S. A. Adsorption of von Willebrand factor/factor VIII by the genetically distinct interstitial collagens. Thromb Res. 1981 Mar 15;21(6):689–691. doi: 10.1016/0049-3848(81)90272-3. [DOI] [PubMed] [Google Scholar]
  39. Santoro S. A., Cunningham L. W. Fibronectin and the multiple interaction model for platelet-collagen adhesion. Proc Natl Acad Sci U S A. 1979 Jun;76(6):2644–2648. doi: 10.1073/pnas.76.6.2644. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Shadle P. J., Barondes S. H. Adhesion of human platelets to immobilized trimeric collagen. J Cell Biol. 1982 Oct;95(1):361–365. doi: 10.1083/jcb.95.1.361. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Silberberg A., Klein J. Structure and properties of surface adsorbed biopolymer layers - soluble collagen adsorbed to glass. Biorheology. 1981;18(3-6):589–599. doi: 10.3233/bir-1981-183-622. [DOI] [PubMed] [Google Scholar]
  42. Sixma J. J., Wester J. The hemostatic plug. Semin Hematol. 1977 Jul;14(3):265–299. [PubMed] [Google Scholar]
  43. Sochynsky R. A., Boughton B. J., Burns J., Sykes B. C., McGee J. O. The effect of human fibronectin on platelet-collagen adhesion. Thromb Res. 1980 May 1;18(3-4):521–533. doi: 10.1016/0049-3848(80)90348-5. [DOI] [PubMed] [Google Scholar]
  44. Stegemann H., Stalder K. Determination of hydroxyproline. Clin Chim Acta. 1967 Nov;18(2):267–273. doi: 10.1016/0009-8981(67)90167-2. [DOI] [PubMed] [Google Scholar]
  45. Szymanowicz A. G., Bellon G., Laurain-Guillaume G., Delvincourt T., Randoux A., Caulet T., Borel J. P. An evaluation by sequential extraction of the proportions of collagen types from medium sized arteries. Artery. 1982;10(4):250–265. [PubMed] [Google Scholar]
  46. Tschopp T. B., Weiss H. J., Baumgartner H. R. Decreased adhesion of platelets to subendothelium in von Willebrand's disease. J Lab Clin Med. 1974 Feb;83(2):296–300. [PubMed] [Google Scholar]
  47. Turitto V. T., Weiss H. J., Baumgartner H. R. Decreased platelet adhesion on vessel segments in von Willebrand's disease: a defect in initial platelet attachment. J Lab Clin Med. 1983 Oct;102(4):551–564. [PubMed] [Google Scholar]
  48. Vuento M., Vaheri A. Purification of fibronectin from human plasma by affinity chromatography under non-denaturing conditions. Biochem J. 1979 Nov 1;183(2):331–337. doi: 10.1042/bj1830331. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Wagner D. D., Urban-Pickering M., Marder V. J. Von Willebrand protein binds to extracellular matrices independently of collagen. Proc Natl Acad Sci U S A. 1984 Jan;81(2):471–475. doi: 10.1073/pnas.81.2.471. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Weiss H. J. Aspirin ingestion compared with bleeding disorders--search for a useful platelet antiaggregant. Blood. 1970 Mar;35(3):333–340. [PubMed] [Google Scholar]
  51. Weiss H. J., Baumgartner H. R., Tschopp T. B., Turitto V. T., Cohen D. Correction by factor VIII of the impaired platelet adhesion to subendothelium in von Willebrand disease. Blood. 1978 Feb;51(2):267–279. [PubMed] [Google Scholar]
  52. Weiss H. J., Turitto V. T., Baumgartner H. R. Effect of shear rate on platelet interaction with subendothelium in citrated and native blood. I. Shear rate--dependent decrease of adhesion in von Willebrand's disease and the Bernard-Soulier syndrome. J Lab Clin Med. 1978 Nov;92(5):750–764. [PubMed] [Google Scholar]
  53. Williams B. R., Gelman R. A., Poppke D. C., Piez K. A. Collagen fibril formation. Optimal in vitro conditions and preliminary kinetic results. J Biol Chem. 1978 Sep 25;253(18):6578–6585. [PubMed] [Google Scholar]
  54. Zucker M. B., Mosesson M. W., Broekman M. J., Kaplan K. L. Release of platelet fibronectin (cold-insoluble globulin) from alpha granules induced by thrombin or collagen; lack of requirement for plasma fibronectin in ADP-induced platelet aggregation. Blood. 1979 Jul;54(1):8–12. [PubMed] [Google Scholar]
  55. van Mourik J. A., Mochtar I. A. Purification of human antihemophilic factor (factor VIII) by gel chromatography. Biochim Biophys Acta. 1970 Dec 22;221(3):677–679. doi: 10.1016/0005-2795(70)90247-3. [DOI] [PubMed] [Google Scholar]

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