Effects of sodium citrate, low molecular weight heparin, and prostaglandin E1 on aggregation, fibrinogen binding, and enumeration of equine platelets.
Abstract: To investigate the effects of sodium citrate, low molecular weight heparin (LMWH), and prostaglandin E1 (PGE1) on aggregation, fibrinogen binding, and enumeration of equine platelets. Methods: Blood samples obtained from 4 Thoroughbreds. Methods: Blood was collected into syringes in the ratio of 9 parts blood:1 part anticoagulant. Anticoagulants used were sodium citrate, LMWH, sodium citrate and LMWH, or 300 nM PGE1/ml of anticoagulant. Platelet aggregation in response to ADP, collagen, and PGE1 was assessed, using optical aggregometry. Platelet activation was evaluated, using flow cytometry, to detect binding of fluorescein-conjugated anti-human fibrinogen antibody. Plasma concentration of ionized calcium was measured, using an ion-selective electrode. Results: Number of platelets (mean +/- SEM) in samples containing LMWH (109.5+/-11.3 x 10(3) cells/microl) was significantly less than the number in samples containing sodium citrate (187.3+/-30.3 x 10(3) cells/microl). Increasing concentrations of sodium citrate resulted in reductions in platelet aggregation and plasma concentration of ionized calcium. Addition of PGE1 prior to addition of an agonist inhibited platelet aggregation in a concentration-dependent manner, whereas addition of PGE1 4 minutes after addition of ADP resulted in partial reversal of aggregation and fibrinogen binding. Conclusions: A high concentration of sodium citrate in blood samples decreases plasma concentration of ionized calcium, resulting in reduced platelet aggregation and fibrinogen binding. Platelets tend to clump in samples collected into LMWH, precluding its use as an anticoagulant. Platelet aggregation and fibrinogen binding can be reversed by PGE1, which may result in underestimation of platelet activation.
Publication Date: 2001-05-01 PubMed ID: 11327462DOI: 10.2460/ajvr.2001.62.547Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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This research article explores the impact of sodium citrate, low molecular weight heparin (LMWH), and prostaglandin E1 (PGE1) on the aggregation, fibrinogen binding, and count of equine platelets. The findings highlight the influence of sodium citrate in reducing platelet aggregation and fibrinogen binding, the unsuitability of LMWH as an anticoagulant due to platelet clumping, and the ability of PGE1 to reverse platelet aggregation and fibrinogen binding.
Methodology
- The study involved blood samples obtained from 4 Thoroughbreds.
- The collected blood was mixed into syringes in the ratio of 9 parts blood:1 part anticoagulant.
- The anticoagulants applied were sodium citrate, LMWH, a combination of sodium citrate and LMWH, or 300 nM PGE1/ml of anticoagulant.
- Platelet aggregation in response to ADP, collagen, and PGE1 was assessed using optical aggregometry – a laboratory technique used for testing the function of platelets.
- Platelet activation was evaluated with flow cytometry, a tool that helped detect the binding of fluorescein-conjugated anti-human fibrinogen antibody.
- Plasma concentration of ionized calcium was measured using an ion-selective electrode.
Results
- The study found that the number of platelets in samples containing LMWH was significantly less than those containing sodium citrate.
- Increasing concentrations of sodium citrate induced reductions in both platelet aggregation and plasma concentration of ionized calcium.
- Adding PGE1 beforehand led to the inhibition of platelet aggregation in a concentration-dependent manner. Additionally, introducing PGE1 four minutes after adding ADP resulted in a partial reversal of aggregation and fibrinogen binding.
Conclusions
- The high concentration of sodium citrate in blood samples reduces plasma concentration of ionized calcium, which brings about a reduction in platelet aggregation and fibrinogen binding.
- The tendency of platelets to clump in samples collected into LMWH suggests its inappropriateness as an anticoagulant.
- PGE1 can reverse platelet aggregation and fibrinogen binding, indicating that it might result in an underestimation of platelet activation.
Cite This Article
APA
Kingston JK, Bayly WM, Sellon DC, Meyers KM, Wardrop KJ.
(2001).
Effects of sodium citrate, low molecular weight heparin, and prostaglandin E1 on aggregation, fibrinogen binding, and enumeration of equine platelets.
Am J Vet Res, 62(4), 547-554.
https://doi.org/10.2460/ajvr.2001.62.547 Publication
Researcher Affiliations
- Department of Veterinary Clinical Sciences, College of Veterinary Medicine, Washington State University, Pullman 99164-6610, USA.
MeSH Terms
- Adenosine Diphosphate / pharmacology
- Alprostadil / pharmacology
- Animals
- Anticoagulants / pharmacology
- Blood Platelets / cytology
- Blood Platelets / drug effects
- Blood Platelets / metabolism
- Calcium / blood
- Citrates / pharmacology
- Fibrinogen / metabolism
- Fibrinolytic Agents / pharmacology
- Flow Cytometry / veterinary
- Hematocrit / veterinary
- Heparin, Low-Molecular-Weight / pharmacology
- Horses / blood
- Platelet Aggregation / drug effects
- Platelet Aggregation / physiology
- Platelet Count / veterinary
- Sodium Citrate
Citations
This article has been cited 9 times.- Gorog DA, Becker RC. Point-of-care platelet function tests: relevance to arterial thrombosis and opportunities for improvement.. J Thromb Thrombolysis 2021 Jan;51(1):1-11.
- Seidel SRT, Vendruscolo CP, Moreira JJ, Fülber J, Ottaiano TF, Oliva MLV, Michelacci YM, Baccarin RYA. Does Double Centrifugation Lead to Premature Platelet Aggregation and Decreased TGF-β1 Concentrations in Equine Platelet-Rich Plasma?. Vet Sci 2019 Aug 21;6(3).
- Mehbuba Hossain S, Chowdhury EH. Citrate- and Succinate-Modified Carbonate Apatite Nanoparticles with Loaded Doxorubicin Exhibit Potent Anticancer Activity against Breast Cancer Cells.. Pharmaceutics 2018 Mar 11;10(1).
- Gutiérrez CM, López C, Giraldo CE, Carmona JU. Study of a Two-Step Centrifugation Protocol for Concentrating Cells and Growth Factors in Bovine Platelet-Rich Plasma.. Vet Med Int 2017;2017:1950401.
- Khorshidi H, Raoofi S, Bagheri R, Banihashemi H. Comparison of the Mechanical Properties of Early Leukocyte- and Platelet-Rich Fibrin versus PRGF/Endoret Membranes.. Int J Dent 2016;2016:1849207.
- Epstein KL, Bergren A, Giguère S, Brainard BM. Cardiovascular, colloid osmotic pressure, and hemostatic effects of 2 formulations of hydroxyethyl starch in healthy horses.. J Vet Intern Med 2014 Jan-Feb;28(1):223-33.
- Watson MG, Lopez JM, Paun M, Jones SA. A novel dynamic layer-by-layer assembled nano-scale biointerface: functionality tests with platelet adhesion and aggregate morphology influenced by adenosine diphosphate.. J Thromb Thrombolysis 2013 Nov;36(4):448-57.
- Solomon C, Winterhalter M, Gilde I, Hoy L, Calatzis A, Rahe-Meyer N. Influence of the sample anticoagulant on the measurements of impedance aggregometry in cardiac surgery.. Med Devices (Auckl) 2008 Jul;1:23-30.
- Giordano A, Meazza C, Salvadori M, Paltrinieri S. Thromboelastometric profiles of horses affected by exercise-induced pulmonary hemorrhages.. Vet Med Int 2010 Sep 30;2010.
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