Matthew W. Hall

Formation of occlusive platelet aggregates in whole blood caused by low concentrations of ADP.

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 &mgr;M) 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.

Hypothermia-induced platelet aggregation and cognitive decline in coronary artery bypass surgery: a pilot study.

Hypothermia-induced platelet aggregation (HIPA) was previously reported in whole blood exposed to synthetic surfaces at 24°-32°C in one-third of normal subjects tested. Cardiopulmonary bypass, conducted with hypothermia, may lead to such aggregation, resulting in microvascular occlusion contributing to cognitive impairment. This pilot study was conducted to explore the relationship between HIPA and cognitive outcome at hospital discharge in patients undergoing coronary artery bypass graft (CABG) surgery as a first step toward a longer-term study. Patients (n=45) undergoing mild to moderate hypothermia (32°-28°8C) during CABG surgery underwent cognitive testing preoperatively and prior to hospital discharge. Tests included: visual and verbal memory, mental processing speed, executive function, language, and intellectual function. HIPA was identified using an in vitro assay in which blood flowing in polyvinychloride tubing was subjected to hypothermia, and platelet aggregates were detected using microscopy and passing the exiting blood through a 20-μm pore filter. Forty-four percent of patients exhibited HIPA. The entire cohort exhibited significant postoperative cognitive decline in verbal memory, mental processing speed and executive function. There was greater cognitive decline in the group with HIPA compared with the group not exhibiting this phenomenon. The patients with HIPA showed significant decline in four of five cognitive measures whereas patients not exhibiting this phenomenon declined in only two of five cognitive measures. HIPA appears to be associated with an added risk of cognitive decline immediately following CABG surgery employing mild to moderate hypothermia. The findings of our study suggest the long-term cognitive effects of hypothermia-induced platelet aggregation need to be explored.

Light-scattering instrument to detect thromboemboli in blood

The characteristics and capabilities of a light-scattering microemboli detector (LSMD) are delineated by detailing its state-of-the-art configuration, by discussing the theoretical and empirical aspects of instrument calibration, and by summarizing various experimental studies that have benefited from this instrument. In the past, thromboembolism, which often results when blood contacts medical devices, has eluded scientific scrutiny due to the absence of instruments that could detect and quantify thromboemboli in circulating blood. More recently, the ability of the LSMD to provide continuous, noninvasive detection of thromboemboli in whole blood (meaning that the LSMD probe does not contact the blood) was exploited in various in vitro and ex vivo models to explore thromboembolic phenomena. Through this work, the LSMD evolved as a sensitive and an economical research tool for the study of thromboembolic phenomena.

In vitro model for studying the effects of hemodynamics on device induced thromboembolism in human blood

Biomateria related thromboembolism is a complex phenomenon, affected by such variables as biomaterial surface chemistry, hemodynamics, and individual donor variations. Thus, isolation of the individual variables would greatly facilitate the understanding and inhibition of this phenomenon. A low volume in vitro model with this potential has been developed, with the initial focus on studying the influence of hemodynamics on thromboembolism (TE) in human blood. Patterned after a larger in vitro model for bovine blood used successfully in our laboratory, the smaller model directed fresh human blood in a single pass through 1/32 inch ID PVC tubing and a flow cell at 3 ml/min. The flow cell consisted of alternating abrupt expansions and contractions of cylindrical tubing that could be modified to study the effects of hemodynamic parameters on TE. Thrombus growth in the flow cell was monitored visually by transillumination microscopy. Emboli from the flow cell were detected continuously by a light-scattering microemboli detector (LSMD), and their strength was assessed by using the constant-pressure filtration (CPF) method. Preliminary studies confirmed the potential of this model. Thrombi were observed visually in the flow cell at sites of high vorticity and at flow separation and reattachment points and were also observed to embolize. Emboli were detected by the LSMD downstream of the flow cell in significantly greater numbers than upstream and were coincident with the embolization of thrombi observed visually. Emboli collected downstream of the flow cell occluded the CPF filters at 50 mm Hg, suggesting that they possessed sufficient strength to occlude microvessels. This model may be used to aid in developing a computer model of thromboembolism, which could subsequently be refined with clinical data.