Hemolysis in clinical blood samples leads to inaccurate assay results and often to the need for repeated blood draws. In vitro experiments were conducted to determine the influence on hemolysis in phlebotomy needles and catheters of pressure difference, cannula diameter, and cannula material. Fresh blood from five human volunteers was forced from a syringe inside a pressurized chamber through 14, 18, and 22 gauge 304 stainless steel needles and polyurethane and Teflon catheters, all 40 mm long. Hemolysis was measured in the samples by a spectrophotometer. It was found that hemolysis increased with increases in pressure difference and cannula diameter and no consistent trend could be identified with regard to cannula material. The pressure differences required for significant hemolysis were above those typical of clinical venipuncture blood draws. While there was substantial variability among individuals, the hemolysis values scaled with exponent S = (t/t0)[(tau/tau0)-1]2, where t is the characteristic duration of shear, t0 is a time constant, tau is the wall shear stress, and tau0 is the wall shear stress threshold below which no hemolysis occurs. A hemolysis threshold including both time and shear stress was also defined for S = constant. The threshold implies that a threshold shear stress exists below which erythrocytes are not damaged for any length of exposure time, but that red cells may be damaged by an arbitrarily short period of exposure to sufficiently large shear stress.

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Minute concentrations of ADP are released when platelets are exposed to shear stress during extracorporeal flow. However, based on current methods, these low concentrations have not been shown to have a significant impact on platelet function. We report here the formation of rigid microaggregates (MA) in response to low concentrations of ADP. A newly developed light scattering whole blood aggregometer (LSWBA) was used to detect an aggregation dose response to ADP (0-2 microM) in heparinized (1.5 U/ml) human blood. Although the LSWBA showed that ADP induced MA were reversible, evidence provided by constant pressure filtration (50 mm Hg) suggested that aggregates existed as rigid particles in the blood for up to 6 minutes. The possible implications of these findings to extracorporeal circulation are discussed.

To observe the dynamics of thromboembolism (TE) in an animal model, a carotid-carotid arterial ex vivo shunt was developed. A coronary stent deployed in a 3.5 mm polyvinylchloride (PVC) tubing segment was used as a model device in the shunt. A light-scattering microemboli detector monitored the embolic content of the blood flowing through the shunt at 50-150 ml/min as determined by a clamp-on ultrasound flow probe. The stent was found to actively develop thrombi and release emboli for 1-3 hours when the activated clotting time (ACT) was maintained between 125 and 150 seconds. The shunt flow rate fluctuated considerably (from 50 to 150 ml/min) depending on the animal's activity. When the time profile of this fluctuating flow rate was super imposed on the time profile of embolization, it was noted that sudden increases in flow rate were associated with incidents of embolization. Statistically, sudden flow rate increases of 100% or more were accompanied by embolic events 95% of the time (p < 0.01). Based on the results of this study, it was postulated that the increased embolization may be due to the fluid forces associated with accelerating flow. To explore this postulate, in vitro studies were conducted to compare the effects of pulsatile flow with steady flow on stent induced TE. Results of this study suggested a significant increase (100%) in both stent thrombosis and embolism during pulsatile flow compared with steady flow.