Original articleA novel modification of the Thrombelastograph assay, isolating platelet function, correlates with optical platelet aggregation☆
Section snippets
Materials
Reptilase (batroxobin maranhao; CenterChem, Norwalk, Ct) and factor XIIIa (Nu-Millennium Research, Manhasset, NY) were obtained separately or as a mixture through Hameoscope Corp (Niles, Ill). Monoject sodium heparin and sodium citrate tubes were obtained from Sherwood Medical (St Louis, Mo). We used the GP IIb/IIIa inhibitors eptifibatide (Integrilin injection; Millennium Pharmaceuticals, Cambridge, MA) and abciximab (ReoPro; Eli Lilly, Indianapolis, Ind). PPACK anticoagulant was purchased
Development of the modified Thromboelastograph assay
Several properties can be measured with the standard Thromboelastograph assay, including time to clot formation, rate of clot formation, and development of MA. The in vitro effect of strong inhibitors of platelet function such as GPIIb/IIIa antagonists can be observed with this standard assay in terms of all the parameters measured, including a reduction in MA.22 To measure platelet activation by other agonists and make the assay sensitive to weaker platelet inhibitors such as NSAIDS or
Discussion
Reptilase and thromboelastography have been used previously to study the interaction of platelets with the fibrin network.29 The development of MA has been shown to depend on platelet activation and to be correlated with clot retraction. However, the MA obtained with reptilase and ADP in the absence of factor XIIIa crosslinking was relatively small and slow to develop compared with a thrombin clot.
In this study, the addition of purified factor XIIIa to a reptilase-generated fibrin network
Acknowledgements
We acknowledge the technical assistance of Carolyn C. Snider, Melinda Coker, and Natalie Hammer, as well as the help of the Knoxville Cardiology Group and catheterization-lab staff in collecting blood samples from patients.
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Supported by the University of Tennessee Anesthesiology Research Fund and the Physicians' Medical Education and Research Foundation, as well as Haemoscope Corp, of which Dr Cohen is chief executive officer. Haemoscope Corp may benefit from these studies.