Effect of heparin on in vitro platelet reactivity in cardiac surgical patients-a comparative assessment by whole blood platelet aggregometry and haemostatometry

1992 ◽  
Vol 66 (6) ◽  
pp. 649-656 ◽  
Author(s):  
L.C.H. John ◽  
G.M. Rees ◽  
I.B. Kovacs
1987 ◽  
Vol 58 (02) ◽  
pp. 744-748 ◽  
Author(s):  
A R Saniabadi ◽  
G D O Lowe ◽  
J C Barbenel ◽  
C D Forbes

SummarySpontaneous platelet aggregation (SPA) was studied in human whole blood at 3, 5, 10, 20, 30, 40 and 60 minutes after venepuncture. Using a whole blood platelet counter, SPA was quantified by measuring the fall in single platelet count upon rollermixing aliquots of citrated blood at 37° C. The extent of SPA increased with the time after venepuncture, with a correlation coefficient of 0.819. The inhibitory effect of dipyridamole (Dipy) on SPA was studied: (a) 10 μM at each time interval; (b) 0.5-100 μM at 3 and 30 minutes and (c) 15 μM in combination with 100 μM adenosine, 8 μM 2-chloroadenosine (2ClAd, an ADP receptor blocker) and 50 μM aspirin. There was a rapid decrease in the inhibitory effect of Dipy with the time after venepuncture; the correlation coefficient was -0.533. At all the concentrations studied, Dipy was more effective at 3 minutes than at 30 minutes after venepuncture. A combination of Dipy with adenosine, 2ClAd or aspirin was a more effective inhibitor of SPA than either drug alone. However, when 15 μM Dipy and 10 μM Ad were added together, the inhibitory effect of Dipy was not increased significantly, suggesting that Dipy inhibits platelet aggregation independent of Ad. The increase in SPA with the time after venepuncture was abolished when blood was taken directly into the anticoagulant containing 5 μM 2ClAd. It is suggested that ADP released from the red blood cells is responsible for the increased platelet aggregability with the time after venepuncture and makes a serious contribution to the artifacts of in vitro platelet function studies.


2006 ◽  
Vol 96 (12) ◽  
pp. 781-788 ◽  
Author(s):  
Andreas Calatzis ◽  
Sandra Penz ◽  
Hajna Losonczy ◽  
Wolfgang Siess ◽  
Orsolya Tóth

SummarySeveral 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.


1994 ◽  
Vol 9 (5) ◽  
pp. 556-558
Author(s):  
J. E. Taylor ◽  
J. J. F. Belch ◽  
I. S. Henderson ◽  
W. K. Stewart

2006 ◽  
Vol 52 (3) ◽  
pp. 383-388 ◽  
Author(s):  
Boris T Ivandic ◽  
Philipp Schlick ◽  
Peter Staritz ◽  
Kerstin Kurz ◽  
Hugo A Katus ◽  
...  

Abstract Background: Inhibition of platelet aggregation by clopidogrel may be insufficient in up to 30% of users. These nonresponders carry an increased risk of cardiovascular events. We reported here a simple assay to study clopidogrel responsiveness. Methods: Electrical impedance aggregometry was performed in diluted whole blood in the presence of 5 and 20 μmol/L ADP. Some samples were incubated with 0.1 mmol/L methyl-S-adenosine monophosphate (MeSAMP), a P2Y12 receptor blocker, to maximize inhibition of aggregation before aggregometry. To validate the assay, we analyzed 6-min impedance in 21 healthy probands and 244 patients with coronary artery disease (CAD). Results: At 5 μmol/L ADP, the imprecision of the assay was 11%. Mean (SD) impedance of the healthy cohort was 12.2 (2.2) Ω. The mean − 3 SD was used to define the cutoff for clopidogrel responsiveness: responders and nonresponders exhibited a 6-min impedance ≤5 Ω and &gt;5 Ω, respectively. Samples from nonresponders were incubated with MeSAMP and analyzed again to distinguish pharmacokinetic and pharmacodynamic types of resistance. Sixteen percent of CAD patients were classified as nonresponders (38 and 2 cases of pharmacokinetic and pharmacodynamic resistance, respectively). Female sex was strongly associated with clopidogrel resistance (P = 0.0002, Fisher exact test). A higher clopidogrel loading dose (P = 0.0353, Mann–Whitney U-test) was given to responders (median, 450 mg) than nonresponders (median, 300 mg). Age and cardiovascular diagnosis showed no significant associations. Conclusions: Impedance aggregometry using 5 μmol/L ADP is a useful tool for studying clopidogrel responsiveness. MeSAMP allows characterization of responsiveness “on treatment” and may be useful for optimizing clopidogrel dosing.


Hematology ◽  
2018 ◽  
Vol 2018 (1) ◽  
pp. 516-521 ◽  
Author(s):  
Rachel Sara Bercovitz

Abstract There is a delicate balance between bleeding and clotting in patients on circuits such as ventricular assist devices or extracorporeal membrane oxygenation. Traditional coagulation tests, prothrombin time, activated partial thromboplastin time, and anti-factor Xa levels, are used to monitor patients on these devices. However, turnaround times and inability to assess global hemostasis, including platelets and fibrinogen have contributed to a recognition that faster, accurate, and more informative coagulation tests are needed. Activated clotting time is used to monitor heparin in patients on circuits and has the advantages of being a near-patient point-of-care test. However, its utility is limited to heparin monitoring. Viscoelastic tests (thromboelastometry and thromboelastography) are global, whole-blood coagulation tests, and whole-blood platelet aggregometry evaluates platelet function. Ideally, these tests can ensure that patients are within the therapeutic range of their antithrombotic medications, identify patients at risk for hemorrhagic or thrombotic complications, and guide management of acute bleeding complications. This ideal is currently hampered by a lack of studies that delineate clear ranges that are clinically relevant. Future research is needed to better understand the optimal use of point-of-care coagulation testing in patients on extracorporeal circuits and ventricular assist devices.


Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 4166-4166
Author(s):  
Neil Blumberg ◽  
Kelly F. Gettings ◽  
Joanna M. Heal

Abstract Three observations led us to investigate whether infusion of ABO non-identical platelets might impair, rather than improve hemostasis. (1) Exposure of platelets to immune complexes or platelet specific antibody can interfere with platelet function in vitro (Thromb Haemost76: 774, 1996). (2) In surgical patients receiving similar numbers of platelet transfusions, those receiving ABO non-identical platelets require 50% more red cell transfusions (Transfusion41: 790, 2001). (3) Patients with acute leukemia receiving prophylactic platelet transfusions typically are reported with serious bleeding at a rate of 15–20%, yet the bleeding rate in patients receiving only ABO identical platelets is below 5% (BMC Blood Disorders4: 6, 2004). In this study, the number of red cell transfusions and clinical outcomes during March 2002-Feb 2003 for all surgical patients of blood groups B and AB (B/AB) who received platelet transfusions were compared with patients of blood groups O and A (O/A). Recipients of blood groups B/AB would not be expected to experience excess bleeding, as measured by red cell transfusions, compared with patients of blood groups O/A. However, because of the lower prevalence of blood groups B/AB in the donor blood supply B/AB recipients may more frequently receive ABO mismatched platelet transfusions. O/A surgical patients (n=281) who received platelet transfusions required a mean of 13 ± 13 (SD) red cell transfusions as compared with B/AB patients (n=54), who required 19 ± 25 red cells (p =0.0086). O/A patients also had shorter length of stay, mean = 25 ± 34 days as compared with B/AB patients at 36 ± 59 days (p =0.064). Rates of mortality and nosocomial infections were not statistically significantly different. O/A patients received a mean of 14 ± 19 units of whole blood platelets during and after surgery, compared with 16 ± 16 units for B/AB patients (p =0.47). O/A patients received a mean of 3.3 ± 6.2 ABO non-identical platelets in contrast with B/AB patients who received 7.5 ± 11 (p = 0.0001). Both groups received similar numbers of ABO identical platelets: 11 ± 16 (O/A) versus 9 ± 12 (B/AB) (p =0.35). All but two patients received only ABO identical FFP and both groups received similar total amounts of FFP (mean of 9 units versus 11 units). While O/A patients received similar mean amounts of cryoprecipitate (6 units) to B/AB patients (8 units), the B/AB patients received significantly more ABO non-identical cryoprecipitate (mean = 4.2 vs. 2.3 units; p = 0.02). To study the effects of ABO incompatible plasma on platelet function, we measured PFA-100 (epinephrine cartridge) closure times in reconstituted whole blood exposing group A platelets to either group A or O plasma. In four of seven instances, closure times for A platelets exposed to O plasma were prolonged by more than 50 seconds, compared with A platelets exposed to allogeneic A plasma. These preliminary results support previous observations that exposure to ABO non-identical platelet transfusions is associated with increased red cell transfusions. One possible mechanism is impaired platelet function caused by antibody or immune complex binding. We speculate that transfusion of ABO mismatched platelets, FFP and/or cryoprecipitate may in some instances exacerbate bleeding, rather than correcting defects in hemostasis. Though further investigation is needed before suggesting changes in clinical practice these findings raise the possibility that use of ABO identical blood components might reduce red cell transfusion needs in bleeding surgical patients.


Author(s):  
Barbara Nunn

The effect of aspirin on human platelet function is usually assessed using platelet-rich plasma (PRP). Some preliminary results in vitro suggested that the effect of aspirin appears to be greater in PRP than whole blood. To explore this possibility further, a comparison of the effect of aspirin in humans ex vivo has been made taking measurements simultaneously in whole blood and PRP at 2 platelet concentrations. Blood samples (36ml) were drawn from 7 male volunteers after a light breakfast. Each took 300mg soluble aspirin and blood samples were drawn again 2 hours later. Blood was mixed with 0.1 volumes 129nM trisodium citrate. Some (30ml) was then centrifuged to prepare PRP and platelet -poor plasma (PPP) by standard techniques. Platelet concentration of some PRP was adjusted with PPP to equal that of the corresponding blood sample; the rest was adjusted to 350,000 per μl. Aggregation in response to collagen (Horm, Munich) was measured photometrically at 37°. Aggregation in 0.5ml aliquots of whole blood was measured after 4 min stirring with 154mM NaCl (control) or collagen at 37° as the fall in single platelet count determined using an Ultraflo- 100 whole blood platelet counter (Clay Adams). The concentrations of collagen producing a 50% maximal response (EC50) in PRP and blood were determined. Dose-ratios for each volunteer were calculated by dividing the EC50 obtained after aspirin by the corresponding value obtained before aspirin.The effect of aspirin was significantly (p<0.001) less in blood than PRP. Whether or not the results in whole blood more closely reflect the effect of aspirin in vivo remains to be determined.


2021 ◽  
Author(s):  
Benjamin Panzer ◽  
Patricia P Wadowski ◽  
Kurt Huber ◽  
Simon Panzer ◽  
Thomas Gremmel

Abstract Background: Dual antiplatelet therapy is a cornerstone in the secondary prevention of ischemic events following percutaneous coronary intervention (PCI) with stent implantation. The new, more potent adenosine diphosphate (ADP) P2Y12 receptor inhibitors prasugrel and ticagrelor have been shown to improve patients’ outcomes. Whether or not these drugs have equal efficacy in diabetic as in non-diabetic individuals is disputed. Furthermore, platelets can be activated by thrombin, which is, at least in part, independent of ADP-inducible activation. Protease-activated receptor (PAR)-1 and -4 are thrombin receptors on human platelets activated by the agonists SFLLRN and AYPGKF, respectively. In the current study, we sought to compare the in vitro efficacy of prasugrel (n=121) and ticagrelor (n=99) to inhibit PAR-mediated platelet activation in patients with type 2 diabetes (n=55).Materials and Methods: We compared P2Y12-, PAR-1- and PAR-4-mediated platelet aggregation as assessed by multiple electrode platelet aggregometry between prasugrel- and ticagrelor-treated patients without and with type 2 diabetes who underwent acute PCI. Results: There were no significant differences of on-treatment platelet aggregation in response to ADP, SFLLRN and AYPGKF between patients on prasugrel or on ticagrelor. Diabetic and non-diabetic patients responded equally. There was no significant correlation between either; ADP-, SFLLRN-, or AYPGKF-inducible platelet aggregation and levels of HbA1c or the body mass index. However, we observed patients with high residual platelet reactivity to SFLLRN and AYPGKF in all cohorts.Conclusion: Prasugrel and ticagrelor inhibit platelet aggregation in diabetic and non-diabetic patients to a similar extent.


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