title: A new method for quantitation of platelet microthrombi and microemboli from cardiopulmonary bypass in organs using 111In labeled platelets.
abstract: During cardiopulmonary bypass (CPB), showers of microemboli (ME) distribute among the organs and connective tissues according to regional blood flow. Post CPB, ME were quantified by subtracting residual platelets (RP) in the organs of a group of unoperated control Yorkshire pigs (n = 6) from those of operated pigs. The RP level was minimized by heparinization (300 IU/kg) before death and exsanguination. The number of adherent microthrombi (MT) and ME from the oxygenator (OX), arterial filter (AF), and thoracotomy site were determined using 111In labeled autologous platelets (INPLT) (525-585 microCi administered 24 hr before CPB) in two CPB groups (ACT > 400 sec) of 12 pigs (30-35 kg). CPB was carried out at a flow of 2.5-3.5 L/min at 28 degrees C with a roller or a centrifugal pump, OX (Bentley Univox 1.8 m2), AF (0.25 m2), and cardiotomy reservoir (CR) (Bentley BR: 3,500), for 90 (n = 6) and 180 (CPB 180, n = 6) min. Six pigs underwent thoracotomy without CPB. L-Arginine was infused at a dose of 2 mg/ kg/min during CPB (n = 6). Flow cytometry was used to estimate the circulating ME in blood. MT and organ trapped ME were imaged with a gamma camera and measured with an ion chamber and a gamma counter. ME values (percent of injected INPLT dose) in six organs and four connective tissues were calculated for all five groups. INPLT distribution indicated a uniform distribution of low level platelet MT in the CR and AF. Circulating ME amounted to 2.5% of total platelets. In the CPB circuit, ME generation in AF was the rate-limiting step (n = 4 x 10(5)). Similar studies in organs and tissues suggested the presence of a uniform distribution of the total events of ME (n = 500 x 10(6)). ME increase in brain, lung, liver, and skeletal muscle following thoracotomy and CPB was significant. The low level of ME in ischemia sensitive organs also indicated the presence of a thrombolytic threshold for cumulative ME. ME disaggregation was activated at an early stage to prevent ischemic damage, specifically in the brain. Measurement of trapped ME provided a novel, reliable, and one step method of evaluation of thrombogenicity of a CPB device and drugs.
authors.affiliation: Department of Veterinary Biosciences, University of Illinois at Champaign-Urbana 61802, USA.
authors.forename: M K
authors.initials: MK
authors.lastname: Dewanjee
authors.forename: P
authors.initials: P
authors.lastname: Zhai
authors.forename: L C
authors.initials: LC
authors.lastname: Hsu
authors.forename: A R
authors.initials: AR
authors.lastname: Twardock
mesh_terms: D000818:Animals; D001698:Biomedical Engineering; D001792:Blood Platelets / Q000000981:diagnostic imaging*; D002315:Cardiopulmonary Bypass / Q000009:adverse effects* / Q000379:methods; D004617:Embolism / Q000000981:diagnostic imaging* / Q000209:etiology*; D005069:Evaluation Studies as Topic; D005434:Flow Cytometry; D015902:Gamma Cameras; D008955:Models, Cardiovascular; D009928:Organ Specificity; D010973:Platelet Adhesiveness; D011877:Radionuclide Imaging; D013552:Swine; D013927:Thrombosis / Q000000981:diagnostic imaging* / Q000209:etiology*