Effects of sodium citrate, low molecular weight heparin, and prostaglandin E1 on aggregation, fibrinogen binding, and enumeration of equine platelets.
OBJECTIVE
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.
Kingston
J K
JK
Department of Veterinary Clinical Sciences, College of Veterinary Medicine, Washington State University, Pullman 99164-6610, USA.
Bayly
W M
WM
Sellon
D C
DC
Meyers
K M
KM
Wardrop
K J
KJ
D000244:Adenosine Diphosphate / Q000494:pharmacology; D000527:Alprostadil / Q000494:pharmacology*; D000818:Animals; D000925:Anticoagulants / Q000494:pharmacology; D001792:Blood Platelets / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D002118:Calcium / Q000097:blood; D002951:Citrates / Q000494:pharmacology*; D005340:Fibrinogen / Q000378:metabolism*; D005343:Fibrinolytic Agents / Q000494:pharmacology; D005434:Flow Cytometry / Q000662:veterinary; D006400:Hematocrit / Q000662:veterinary; D006495:Heparin, Low-Molecular-Weight / Q000494:pharmacology*; D006736:Horses / Q000097:blood*; D010974:Platelet Aggregation / Q000187:drug effects* / Q000502:physiology; D010976:Platelet Count / Q000662:veterinary; D000077559:Sodium Citrate
D000925:Anticoagulants; D002951:Citrates; D005343:Fibrinolytic Agents; D006495:Heparin, Low-Molecular-Weight; D000077559:Sodium Citrate; D000244:Adenosine Diphosphate; D005340:Fibrinogen; D000527:Alprostadil; D002118:Calcium