In Vitro Comparison of Blood Pump Induced Platelet Microaggregates between a Centrifugal and Roller Pump During Cardiopulmonary Bypass

Abstract

Platelets are consumed during cardiopulmonary bypass (CPB) and mechanical ventricular assistance, at least partly as a result of the formation of platelet microaggregates in the blood pump. There is no commonly accepted method currently available to detect platelet microaggregates during the use of CPB or left ventricular assist devices (LVAD). The purpose of this study was to develop a flow cytometric method for the quantification of platelet microaggregates generated in blood pumps, and to evaluate the effect of cellular fragments from hemolyzed erythrocytes on the perioperative assessment of platelet counts during CPB. Method Fresh human anticoagulated blood (1IU heparin /mL, activated clotting time 250 ± 24 sec.) was circulated for 120 minutes in an artificial circulatory system, containing either a centrifugal pump (CP) or roller pump (RP). Whole blood was used to quantify platelet consumption and to detect circulating platelet microaggregates in a flow cytometer. Platelet consumption was additionally analyzed using an automated “Coulter” blood cell counter. Hemolysis was analyzed by measurement of plasma free hemoglobin (fHb), as well a by flow cytometric detection of red blood cell (RBC) fragments. Results Flow cytometric analysis demonstrated significantly more circulating platelet aggregates and platelet consumption in the RP than in the CP (p<0.01). Quantification of RBC fragments and plasma free hemoglobin (fHb) levels also indicated significantly increased hemolysis in the RP than in the CP (p<0.01). In contrast, the Coulter count data indicated less platelet consumption in the PP compared to the CP. Conclusion Fragments from hemolyzed erythrocytes have the same size distribution as intact platelets and the number of RBC fragments correlates with the extent of pump-induced hemolysis during CPB. Our data suggest that assessment of platelets by “Coulter counting” cannot distinguish platelets from RBC fragments and may underestimate platelet consumption in the presence of hemolysis during CPB. We conclude that flow cytometry is more accurate in the perioperative assessment of platelet count and platelet aggregation during CPB and LVAD support.