Multiple electrode aggregometry: A new device to measure platelet aggregation in whole blood

Orsolya Tóth; Andreas Calatzis; Sandra Penz; Hajna Losonczy; Wolfgang Siess

doi:10.1160/TH06-05-0242

Thrombosis and Haemostasis, Inhaltsverzeichnis

Thromb Haemost 2006; 96(06): 781-788
DOI: 10.1160/TH06-05-0242

Platelets and Blood Cells

Schattauer GmbH

Multiple electrode aggregometry: A new device to measure platelet aggregation in whole blood

Autoren

Orsolya Tóth

¹Institute for Prevention of Cardiovascular Diseases, University of Munich, Munich, Germany

²1^stDepartment of Medicine, University of Pécs, Pécs, Hungary
Andreas Calatzis

³Department of Transfusion Medicine and Hemostaseology, University of Munich, Munich, Germany
Sandra Penz

¹Institute for Prevention of Cardiovascular Diseases, University of Munich, Munich, Germany
Hajna Losonczy

²1^stDepartment of Medicine, University of Pécs, Pécs, Hungary
Wolfgang Siess

¹Institute for Prevention of Cardiovascular Diseases, University of Munich, Munich, Germany

Abstract

Summary

Several methods are used to analyse platelet function in whole blood. A new device to measure whole blood platelet aggregation has been developed, called multiple electrode platelet aggregometry (MEA). Our aim was to evaluate MEA in comparison with the single platelet counting (SPC) method for the measurement of platelet aggregation and platelet inhibition by aspirin or apyrase in diluted whole blood. Platelet aggregation induced by different concentrations of ADP, collagen and TRAP-6 and platelet inhibition by apyrase or aspirin were determined in citrateor hirudin-anticoagulated blood by MEA and SPC. MEA indicated that spontaneous platelet aggregation was lower, and stimulated platelet aggregation was higher in hirudin- than citrate-anticoagulated blood. In hirudin-anticoagulated, but not citrate-anticoagulated blood, spontaneous platelet aggregation measured by MEA was inhibited by apyrase. For MEA compared with SPC the dose response-curves of agonist-induced platelet aggregation in citrate- and hirudin-blood showed similar EC50 values for TRAP, and higher EC50 values for ADP (non-significant) and collagen (p<0.05). MEA and the SPC method gave similar results concerning platelet-inhibition by apyrase and aspirin. MEA was more sensitive than SPC to the inhibitory effect of aspirin in collagen-induced aggregation. In conclusion, MEA is an easy, reproducible and sensitive method for measuring spontaneous and stimulated platelet aggregation, and evaluating antiplatelet drugs in diluted whole blood. The use of hirudin as an anticoagulant is preferable to the use of citrate. MEA is a promising technique for experimental and clinical applications.

Keywords

Anticoagulation - multiple electrode aggregometry - platelet aggregation - single platelet counting

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References
1 Born GV, Cross MJ. Effect of adenosine diphosphate on the concentration of platelets in circulating blood. Nature 1963; 197: 974-6.
2 Bartlett GR. Metabolism by man of intravenously administered adenine. Transfusion 1977; 17: 367-73.
3 Santos MT, Valles J, Marcus AJ. et al. Enhancement of platelet reactivity and modulation of eicosanoid production by intact erythrocytes. A new approach to platelet activation and recruitment. J Clin Invest 1991; 87: 571-80.
4 Persidsky MD, Ling NS. Separation of platelet-rich plasma by modified centrifugal elutriation. J Clin Apher 1982; 01: 18-24.
5 Reiss RF, Katz AJ. Optimizing recovery of platelets in platelet rich plasma by the simplex strategy. Transfusion 1976; 16: 370-4.
6 Hill RJ, Stenberg PE, Sullam PM. et al. Use of arabinogalactan to obtain washed murine platelets free of contaminating plasma proteins and appropriate for studies of function, morphology, and thrombopoiesis. J Lab Clin Med 1988; 111: 73-83.
7 Dyszkiewicz-Korpanty AM, Frenkel EP, Sarode R. Approach to the assessment of platelet function: comparison between optical-based platelet-rich plasma and impedance-based whole blood platelet aggregation methods. Clin Appl Thromb Hemost 2005; 11: 25-35.
8 Fox SC, Burgess-Wilson M, Heptinstall S. et al. Platelet aggregation in whole blood determined using the Ultra-Flo 100 Platelet Counter. Thromb Haemost 1982; 48: 327-9.
9 Mandal S, Sarode R, Dash S. et al. Hyperaggregation of platelets detected by whole blood platelet aggregometry in newly diagnosed noninsulin-dependent diabetes mellitus. Am J Clin Pathol 1993; 100: 103-7.
10 Saniabadi AR, Lowe GD, Forbes CD. et al. Platelet aggregation studies in whole human blood. Thromb Res 1983; 30: 625-32.
11 Gray RP, Hendra TJ, Patterson DL. et al. “Spontaneous” platelet aggregation in whole blood in diabetic and non diabetic survivors of acute myocardial infarction. Thromb Haemost 1993; 70: 932-6.
12 Cho NH, Becker D, Dorman JS. et al. Spontaneous whole blood platelet aggregation in insulin-dependent diabetes mellitus: an evaluation in an epidemiologic study. Thromb Haemost 1989; 61: 127-30.
13 Haseruck N, Erl W, Pandey D. et al. The plaque lipid lysophosphatidic acid stimulates platelet activation and platelet-monocyte aggregate formation in whole blood: involvement of P2Y1 and P2Y12 receptors. Blood 2004; 103: 2585-92.
14 Cardinal DC, Flower RJ. The electronic aggregometer: a novel device for assessing platelet behavior in blood. J Pharmacol Methods 1980; 03: 135-58.
15 Riess H, Braun G, Brehm G. et al. Critical evaluation of platelet aggregation in whole human blood. Am J Clin Pathol 1986; 85: 50-6.
16 Ingerman-Wojenski C, Smith JB, Silver MJ. Evaluation of electrical aggregometry: comparison with optical aggregometry, secretion of ATP, and accumulation of radiolabeled platelets. J Lab Clin Med 1983; 101: 44-52.
17 Wallen NH, Ladjevardi M, Albert J. et al. Influence of different anticoagulants on platelet aggregation in whole blood; a comparison between citrate, low molecular mass heparin and hirudin. Thromb Res 1997; 87: 151-7.
18 Phillips DR, Teng W, Arfsten A. et al. Effect of Ca2+ on GP IIb-IIIa interactions with integrilin: enhanced GP IIb-IIIa binding and inhibition of platelet aggregation by reductions in the concentration of ionized calcium in plasma anticoagulated with citrate. Circulation 1997; 96: 1488-94.
19 Storey RF, Wilcox RG, Heptinstall S. Differential effects of glycoprotein IIb/IIIa antagonists on platelet microaggregate and macroaggregate formation and effect of anticoagulant on antagonist potency. Implications for assay methodology and comparison of different antagonists. Circulation 1998; 98: 1616-21.
20 Sharp DS, Beswick AD, O’Brien JR. et al. The association of platelet and red cell count with platelet impedance changes in whole blood and light-scattering changes in platelet rich plasma: evidence from the Caerphilly Collaborative Heart Disease Study. Thromb Haemost 1990; 64: 211-5.
21 Armstrong R, May JA, Losche W. et al. Factors that contribute to spontaneous platelet aggregation and streptokinase-induced aggregation in whole blood. Thromb Haemost 1995; 73: 297-303.
22 Sathiropas P, Marbet GA, Sahaphong S. et al. Detection of small inhibitory effects of acetylsalicylic acid (ASA) by platelet impedance aggregometry in whole blood. Thromb Res 1988; 51: 55-62.
23 Storey RF, Newby LJ, Heptinstall S. Effects of P2Y(1) and P2Y(12) receptor antagonists on platelet aggregation induced by different agonists in human whole blood. Platelets 2001; 12: 443-7.