Download PDF
Thromb Haemost 1990; 63(01): 112-121
DOI: 10.1055/s-0038-1645696
Original Article
Schattauer GmbH Stuttgart

The Effect of Red Blood Cells on the ADP-induced Aggregation of Human Platelets in Flow Through Tubes

David N Bell
The McGill University Medical Clinic, Montreal General Hospital, and the Department of Medicine, McGill University, Montreal, Canada
,
Samira Spain
The McGill University Medical Clinic, Montreal General Hospital, and the Department of Medicine, McGill University, Montreal, Canada
,
Harry L Goldsmith
The McGill University Medical Clinic, Montreal General Hospital, and the Department of Medicine, McGill University, Montreal, Canada
› Author Affiliations
Further Information

Publication History

Received 20 January 1989

Accepted after revision23 October 1989

Publication Date:
02 July 2018 (online)

Also available at
  PDF Download  Permissions and Reprints

Summary

The effect of red blood cells, rbc, and shear rate on the ADPinduced aggregation of platelets in whole blood, WB, flowing through polyethylene tubing was studied using a previously described technique (1). Effluent WB was collected into 0.5% glutaraldehyde and the red blood cells removed by centrifugation through Percoll. At 23°C the rate of single platelet aggregtion was upt to 9× greater in WB than previously found in platelet-rich plasma (2) at mean tube shear rates Ḡ = 41.9,335, and 1,920 s−1, and at both 0.2 and 1.0 µM ADP. At 0.2 pM ADP, the rate of aggregation was greatest at Ḡ = 41.9 s−1 over the first 1.7 s mean transit time through the flow tube, t, but decreased steadily with time. At Ḡ ≥335 s−1 the rate of aggregation increased between t = 1.7 and 8.6 s; however, aggregate size decreased with increasing shear rate. At 1.0 µM ADP, the initial rate of single platelet aggregation was still highest at Ḡ = 41.9 s1 where large aggregates up to several millimeters in diameter containing rbc formed by t = 43 s. At this ADP concentration, aggregate size was still limited at Ḡ ≥335 s−1 but the rate of single platelet aggregation was markedly greater than at 0.2 pM ADP. By t = 43 s, no single platelets remained and rbc were not incorporated into aggregates. Although aggregate size increased slowly, large aggregates eventually formed. White blood cells were not significantly incorporated into aggregates at any shear rate or ADP concentration. Since the present technique did not induce platelet thromboxane A2 formation or cause cell lysis, these experiments provide evidence for a purely mechanical effect of rbc in augmenting platelet aggregation in WB.

Keywords

Platelet aggregation - Shear rate - Red blood cells
 

  • References

  • 1 Bell DN, Spain S, Goldsmith HL. The ADP-induced aggregation of human platelets in flow through tubes: I Measurement of the concentration and size of single platelets and aggregates. Biophys J 1989; 56: 817-828
    CrossrefPubMedGoogle Scholar
  • 2 Bell DN, Spain S, Goldsmith HL. The ADP-induced aggregation of human platelets in flow through tubes: II. Effect of shear rate, donor sex, and ADP concentration. Biophys J 1989 56. 829-843
    PubMedGoogle Scholar
  • 3 Begent N, Born GV R. Growth rate in vivo of platelet thrombi, produced by iontophoresis of ADP, as a function of mean blood flow velocity. Nature 1970; 227: 926-930
    CrossrefPubMedGoogle Scholar
  • 4 Arfors K-E, Cockburn JS, Gross JF. Measurement of growth rate of laser-induced intravascular platelet aggregation and the influence of blood flow velocity. Microvasc Res 1976; 11: 79-87
    CrossrefPubMedGoogle Scholar
  • 5 McPherson VJ, Zucker MB, Friedberg NM, Rifkin PL. Platelet retention in glass bead columns: further evidence for the importance of ADP. Blood 1974; 44: 411-425
    PubMedGoogle Scholar
  • 6 Fox S, Burgess-Wilson ME, Heptinstall S, Mitchell JR. Platelet aggregation in whole blood determined using the Ultra-Flo 100 platelet counter. Thromb Haemostas 1982; 48: 327-329
    PubMedGoogle Scholar
  • 7 Saniabadi AR, Lowe GD O, Barbenel JC, Forbes CD. A comparison of spontaneous platelet aggregation in whole blood with platelet rich plasma: additional evidence for the role of ADP. Thromb Haemostas 1984; 51: 115-118
    PubMedGoogle Scholar
  • 8 Jen CJ, Mclntire LV. Characteristics of shear-induced aggregation in whole blood. J Lab Clin Med 1984; 103: 115-124
    PubMedGoogle Scholar
  • 9 Zawilska KM, Born GV R, Begent NA. Effect of ADP-utilizing enzymes on the arterial bleeding time in rats and rabbits. Br J Haematol 1982; 50: 317-325
    CrossrefPubMedGoogle Scholar
  • 10 Skoza L, Zucker MB, Jerushalmy Z, Grant R. Kinetic studies of aggregation induced by adenosine diphosphate and its inhibition by chelating agents, guanidino compounds, and adenosine. Thromb Diath Haemorrh 1967; 18: 713-725
    PubMedGoogle Scholar
  • 11 Parker JC. Metabolism of external adenine nucleotides by human red blood cells. Am J Physiol 1970; 218: 1568-1574
    PubMedGoogle Scholar
  • 12 Gresele P, Arnout J, Deckmyn H, Vermylen J. Mechanism of the antiplatelet action of dipyridamole in whole blood: modulation of adenosine concentration and activity. Thromb Haemostas 1986; 55: 12-18
    PubMedGoogle Scholar
  • 13 Barker LA, Kadatz RA. Mechanism of action of dipyridamole. Thromb Res 1983; (Suppl. 04) 39-46
    PubMedGoogle Scholar
  • 14 Gresele P, Zoja C, Deckmyn H, Arnout J, Vermylen J, Verstraete M. Dipyridamole inhibits platelet aggregation in whole blood. Thromb Haemost 1983; 50: 852-856
    Article in Thieme ConnectPubMedGoogle Scholar
  • 15 Holmsen H, Day HJ, Strom E. Adenine nucleotide metabolism of blood platelets. VI. Subcellular localization of nucleotide pools with different functions in the platelet release reaction. Biochim Biophys Acta 1969 186. 254-266
    PubMedGoogle Scholar
  • 16 Gresele P, Arnout J, Deckmyn H, Vermylen J. Combining antiplatelet agents: potentiation between aspirin and dipyridamole. Lancet 1985; 1: 937-938
    PubMedGoogle Scholar
  • 17 Riess H, Braun G, Brehn G, Hiller E. Critical evaluation of platelet aggregation in whole human blood. Am J Clin Pathol 1986; 85: 50-56
    CrossrefPubMedGoogle Scholar
  • 18 Goldsmith HL. Red cell motions and wall interactions in tube flow. Fed Proc 1971; 30: 1578-1588
    PubMedGoogle Scholar
  • 19 Turitto VT, Benis AM, Leonard EF. Platelet diffusion in flowing blood.. lnd Eng Chem Fundam 1972; 11: 216-223
    PubMedGoogle Scholar
  • 20 Tangelder GJ, Teirlinck HC, Slaaf DW, Reneman RS. Distribution of blood platelets in flowing arterioles. Am J Physiol 1985; 248: H318-h323
    PubMedGoogle Scholar
  • 21 Corattiyl V, Eckstein EC. Regional platelet concentration in blood flow through capillary tubes. Microvasc Res 1986; 32: 261-270
    CrossrefPubMedGoogle Scholar
  • 22 Zucker MB, Rifkin PL, Friedberg NM, Coller BS. Mechanism of platelet function as revealed by the retention of platelets in glass bead columns. Ann N Y Acad Sci 1972; 201: 138-144
    CrossrefPubMedGoogle Scholar
  • 23 Karino T, Goldsmith HL. Adhesion of human platelets to collagen on the walls distal to a tubular expansion. Microvasc Res 1979; 17: 238-262
    CrossrefPubMedGoogle Scholar
  • 24 Reimers RC, Sutera SP, Joist JH. Potentiation by red blood cells of shear-induced platelet aggregation: relative importance of chemical and physical mechanisms. Blood 1984; 64: 1200-1206
    CrossrefPubMedGoogle Scholar
  • 25 Bell DN, Spain S, Goldsmith HL. Extracellular free Ca2+ accounts for the sex difference in the aggregation of human platelets in citrated platelet-rich plasma. Thromb Res: submitted
    PubMedGoogle Scholar
  • 26 Bell DN. Physical factors governing the aggregation of human platelets in sheared suspensions. Ph. D. Thesis, McGill University, Montreal, Quebec 1988
    PubMedGoogle Scholar
  • 27 Goldsmith HL, Marlow JC. Flow behavior of erythrocytes. II.Particle motions in sheared suspensions of ghost cells. J Colloid Interface Sci 1979 71. 383-407
    PubMedGoogle Scholar
  • 28 Dittmer DS. ed Blood and Other Body Fluids. Federation of American Society for Experimental Biology; Washington, DC: 1961: 102
    Google Scholar
  • 29 Grover NB, Naaman N, Ben-Sasson S, Doljanski F. Electrical sizing of particles in suspensions. I. Theory. Biophys J 1969 9. 1398-415
    PubMedGoogle Scholar
  • 30 Nakeff A, Ingram M. Platelet count : volume relationships in four mammalian species. J Appl Physiol 1970; 28: 530-533
    CrossrefPubMedGoogle Scholar
  • 31 Paulus J. Platelet size in man. Blood 1975; 46: 321-336
    PubMedGoogle Scholar
  • 32 Mundschenk DD, Connelly DP, White JG, Brunning RD. An improved technique for the electronic measurement of platelet size and shape. J Lab Clin Med 1976; 88: 301-315
    PubMedGoogle Scholar
  • 33 Holme S, Murphy S. Coulter Counter and light transmission studies of platelets exposed to low temperature, ADP, EDTA, and storage at 22°. J Lab Clin Med 1980; 96: 480-493
    PubMedGoogle Scholar
  • 35 Bahr GF, Zeitler E. The determination of the dry mass in populations of isolated particles. Lab Invest 1965; 14: 955-977
    PubMedGoogle Scholar
  • 35 Von Behrens WE. Platelet size. M. D. Thesis, University of Adelaide, Adelaide, S. A 1972
    PubMedGoogle Scholar
  • 36 Nichols AR, Bosman HB. Platelet aggregation: newly quantified using nonempirical parameters. Thromb Haemostas 1979; 42: 679-93
    PubMedGoogle Scholar
  • 1 Gear AR L. Rapid reactions of platelets studied by a quenched-flow approach: Aggregation kinetics. J Lab Clin Med 1982; 100: 866-886
    PubMedGoogle Scholar
  • 38 Belval T, Heliums JD. The analysis of shear-induced platelet aggregation with population balance mathematics. Biophys J 1986; 50: 479-487
    CrossrefPubMedGoogle Scholar
  • 39 Valdorf-Hansen JF, Zucker MB. Effect of temperature and inhibitors on serotonin-14C release from human platelets. Am J Physiol 1971; 220: 105-111
    PubMedGoogle Scholar
  • 40 Mustard JF, Perry DW, Kinlough-Rathbone RL, Packham MA. Factors responsible for ADP-induced release reaction of human platelets. Am J Physiol 1975; 228: 1757-1765
    PubMedGoogle Scholar
  • 41 Lages B, Weiss HJ. Dependence of human functional responses on divalent cations: Aggregation and secretion in heparin- and hirudinanticoagulated platelet-rich plasma and the effects of chelating agents. Thromb Haemostas 1981; 45: 173-179
    PubMedGoogle Scholar
  • 42 Bell DN, Goldsmith HL. Platelet aggregation in Poiseuille flow: II. Effect of shear rate. Microvasc Res 1984 27. 316-330
    PubMedGoogle Scholar
  • 43 Goldsmith HL, Mason SG. The microrheology of dispersions. In: Rheology: Theory and Applications Eirich FR. ed 4 Academic Press; New York: 1967: 85-250
  • 44 Van de Ven TG M, Mason SG. The microrheology of colloidal dispersions. VII. Orthokinetic doublet formation of spheres. Coll Polymer Sci 1977 255. 468-479
    PubMedGoogle Scholar
  • 45 Belval T, Heliums JD. The analysis of shear-induced platelet aggregation with population balance mathematics. Biophys J 1986; 50: 479-1029
    CrossrefPubMedGoogle Scholar
  • 46 Turitto VT, Baumgartner HR. Platelet interaction with subendothelium in a perfusion system: physical role of red blood cells. Microvasc Res 1975; 9: 335-44
    CrossrefPubMedGoogle Scholar
  • 1 Smoluchowski Mvon. Versuch einer mathematischen Theorie der Koagulationskinetik kolloider Losungen. Z Physik Chem 1917; 92: 129-68
    PubMedGoogle Scholar
  • 48 Brown CH, Leverett LB, Lewis CW, Alfrey CP, Heliums JD. Morphological, biochemical, and functional changes in human blood platelets subjected to shear stress. J Lab Clin Med 1975; 86: 462-471
    PubMedGoogle Scholar
  • 49 Dewitz TS, Martin RR, Solis RT, Heliums J, McIntyre LV. Microaggregate formation in whole blood exposed to shear stress. Microvasc Res 1978; 16: 263-71
    CrossrefPubMedGoogle Scholar
  • 50 Frojmovic MM, Milton JG. Human platelet size, shape and related functions in health and disease. Physiol Rev 1982; 62: 185-261
    CrossrefPubMedGoogle Scholar
  • 51 Milton JG, Frojmovic MM. Adrenaline and ADP-induced platelet aggregation require shape change: importance of pseudopods. J Lab Clin Med 1984; 104: 805-815
    PubMedGoogle Scholar
  • 52 Hellem AJ. The adhesiveness of human blood platelets in vitro. Scand J Clin Lab Invest 1960; 12 (Suppl. 51) 1-117
    CrossrefPubMedGoogle Scholar
  • 53 Born GV R, Wchmcier A. Inhibition of platelet thrombus formation by chlorpromazine acting to diminish haemolysis. Nature 1979; 282: 212-3
    CrossrefPubMedGoogle Scholar
  • 54 Leverett LB, Heliums JD, Alfrey CP, Lynch EC. Red blood cell damage by shear stress. Biophys J 1972; 12: 257-273
    CrossrefPubMedGoogle Scholar
  • 55 Heliums JD, Hardwick RA. Response of platelets to shear stress – a review. In The Rheology of Blood, Blood Vessels and Associated Tissues. Gross DR, Hwang NH C. eds Sijthoff and Noordhoff; Rockville, MD: 1981: 160-183
  • 56 Burgess-Wilson ME, Green S, Heptinstall S, Mitchell JR A. Spontaneous platelet aggregation in whole blood: dependence on age and haematocrit. Lancet 1984; 2: 1213
    PubMedGoogle Scholar
  • 57 Saniabadi A, Lowe G DO, Barbenel JC, Forbes CD. Further studies on the role of red blood cells in spontaneous platelet aggregation. Thromb Res 1985; 38: 225-232
    CrossrefPubMedGoogle Scholar
  • 58 Abbate R, Favilla S, Boddi M, Costanzo G, Prisco D. Factors influencing platelet aggregation in whole blood. Am J Clin Pathol 1986; 86: 91-96
    CrossrefPubMedGoogle Scholar
  • 59 Breddin K, Grun H, Krzywanck HJ, Schremmer WP. On the measurement of spontaneous platelet aggregation. The platelet aggregation test III. Methods and first clinical results. Thromb Haemostas 1976; 35: 669-691
    PubMedGoogle Scholar
  • 60 Adams GA, Feuerstein IA. Platelet adhesion and release: Interfacial concentration of released materials. Am J Physiol 1981; 240: H99-H108
    PubMedGoogle Scholar