Abstract 391: Device-Relevant Dynamic and Constant Shear Stresses Induce Prothrombotic Platelet- and Monocyte-Derived Microparticles

Abstract

Cardiovascular blood-recirculating devices provide life saving cardiopulmonary support to critically ill populations. Unfortunately, common device-related thrombotic complications continue to result in significant mortality, neurological morbidity, and limb loss. Past studies have shown that increased wall shear stress from blood-recirculating devices activates circulating monocytes and platelets. Less well studied is the enhancement of thrombosis by platelet-derived (PMPs) and monocyte-derived microparticles (MoMPs). These are small (0.1-1 micron) cell-derived membrane vesicles released from activated cells. Studies using a biological stimulant demonstrate that MoMPs initiate clotting whereas PMPs enhance clotting. Few if any studies have defined the contributions of device-produced microparticles (MPs) to changes in thrombotic potential. Our hypothesis is the increased dynamic or constant shear stress generated by a blood-recirculating device promotes release of prothrombotic MPs. Using an Anton Paar MCR Rheometer we exposed whole human blood in vitro to both dynamic (70 dynes/cm 2 at 0.05hz) and constant (70 and 35 dynes/cm 2 ) shear stress conditions for two minutes. After shear, MPs were collected with dual centrifugation and then quantified using a BD FACS Canto II flow cytometer with forward scatter photomultiplier tube (resolution=200nm). Then we pelleted and washed MPs to measure changes in thrombin generation with a Calibrated Automated Thrombogram (CAT). Dynamic shear stress generated 3841±1171 PMPs/μL and 3522±1090 MoMPs/μL, (MEAN±SE, n=6). Constant shear stress at 70 dynes/cm 2 and 35 dynes/cm 2 generated PMPs/μL of 4937±1655 and 1880±389 and MoMPs/μL of 6935±2029 (n=3) and 2262±1041 respectively (MEAN±SE, n=3 for each). All shear conditions generated an increase in MP concentration over static conditions of 644±120 PMPs/μL and 538 MoMPs/μL (p<0.01). Dynamic and constant sheared MPs increased the estimated thrombin potential and peak thrombin generation by at least 30% compared to static (p<0.05). This study is the first to demonstrate that device relevant constant and dynamic shear stresses can generate PMPs and MoMPs that promote thrombosis.