fibrinopeptide-a and Hemolysis

An in vitro model cardiopulmonary bypass (CPB) circuit consisting ot tubing, oxygenator and venous reservoirs with either a roller or a centrifugal pump, and with either heparin-coated (Carmeda Bioactive Surface, CBAS) or uncoated surfaces, was studied with respect to 'blood activation', using small-scale-based blood volume (450 + 500 ml). Sixteen circuits were tested in each pump group, eight with and eight without heparin-coated surfaces, by circulating heparinized fresh human blood for 72 hours at 30 degrees C. Blood plasma, sampled at defined intervals, was analysed for haemolysis (lactate dehydrogenase and potassium), complement activation (C3bc and C5b-9 (TCC)), complement lytic inhibitors (vitronectin and clusterin), coagulation activation (fibrinopeptide A), granulocyte (lactoferrin and myeloperoxidase) and platelet (beta-thromboglobulin) activation and contaminating endotoxin. The heparin coating significantly reduced the concentrations of C3bc, TCC, fibrinopeptide A, lactoferrin, myeloperoxidase and beta-thromboglobulin. The two pump types did not differ with respect to these parameters, but the roller pump caused significantly higher increases in plasma LDH and potassium and significantly greater reductions in clusterin and vitronectin than the centrifugal pump. Endotoxin concentration was low at the start and after 24 hours in all groups. These results confirm that heparin-coated CPB surfaces reduce blood activation, and suggest that centrifugal pumps cause less haemolysis and less reduction in lytic complement inhibitors than roller pumps.

Topics: Blood Coagulation; Cardiopulmonary Bypass; Complement Activation; Complement Inactivator Proteins; Extracorporeal Circulation; Fibrinopeptide A; Granulocytes; Hemolysis; Heparin; Humans; L-Lactate Dehydrogenase; Oxygenators, Membrane; Platelet Activation; Surface Properties

Centrifugal pumps are being used increasingly for short-term extracorporeal circulation purposes such as during heart operations. Whether the centrifugal pump improves the cardiopulmonary bypass biocompatibility has not been fully documented.. A roller pump (n = 20) was compared in vivo with a centrifugal pump (n = 20) in groups of patients in which cardiopulmonary bypass circuits that were either totally heparin coated (Carmeda BioActive Surface; n = 20) or uncoated (n = 20) were used. We expected the heparin coating to attenuate blood activation, thus possibly making the comparison of the two pumps easier with respect to their different blood activation potentials. Samples of blood plasma, obtained during cardiopulmonary bypass from low-risk coronary artery bypass grafting patients, were analyzed for hemolysis (plasma haemoglobin), complement activation (C3bc and the terminal complement complex), a complement lytic inhibitor (vitronectin), coagulation activation (fibrinopeptide A), granulocyte activation (lactoferrin), and platelet activation (beta-thromboglobulin).. The concentrations of terminal complement complex, lactoferrin, and beta-thromboglobulin were significantly lower in association with heparin-coated surfaces. The concentration of plasma hemoglobin was significantly lower in association with the centrifugal pump. In uncoated circuits, the beta-thromboglobulin level was significantly higher in association with the roller pump than with the centrifugal pump, but this significant reduction in the beta-thromboglobulin level did not hold true for the heparin-coated circuit group.. A heparin-coated cardiopulmonary bypass surface reduces the blood activation potential during cardiopulmonary bypass, and the centrifugal pump causes less hemolysis than the roller pump.

Topics: Adult; Aged; beta-Thromboglobulin; Biocompatible Materials; Cardiopulmonary Bypass; Complement C3b; Complement Membrane Attack Complex; Coronary Artery Bypass; Female; Fibrinopeptide A; Hemoglobins; Hemolysis; Heparin; Humans; Lactoferrin; Male; Middle Aged; Vitronectin

A new, automated technique for the preparation of blood components is described. A system of 3 or 4 integrally connected plastic containers (Optipac) is handled by a new type of extractor (Optipress). The container in which the blood is collected has an outlet at the top and another at the bottom. After normal centrifugation to obtain separation of the blood components, these are squeezed out from the top and bottom simultaneously under control of a photocell. The primary separation step results in three components: a leukocyte-poor red-cell suspension in SAGM medium, CPD plasma, and a buffy-coat preparation. The system has been tested in two laboratories (lab A and lab B). A 'heavy-spin' centrifugation to obtain a maximum yield of cell-poor plasma gave the best removal of leukocytes from the red cells; the remaining leukocyte content was 0.46 +/- 0.25 (lab A) and 0.5 +/- 0.4 (lab B) x 10(9)/red-cell unit. Platelet concentrates can be prepared either the normal way via platelet-rich plasma or from buffy coat. Red-cell 24-hour autologous posttransfusion survival using labeling with 51Cr was 87.5 +/- 4.1% (lab A) after 35 days, and 84.2 +/- 4.2% (lab A) and 77.5 +/- 1.5% (lab B) after 42 days. Red-cell morphology and fluidity compared favorably to previous studies using the same additive solution in traditional plastic-bag systems. The total adenine nucleotide concentration was maintained normal for 42 days.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: 2,3-Diphosphoglycerate; Adenosine Monophosphate; Blood Cell Count; Blood Glucose; Blood Preservation; Diphosphoglyceric Acids; Erythrocyte Aging; Erythrocytes; Fibrinopeptide A; Hemolysis; Humans; Hydrogen-Ion Concentration; Kallikreins; Peptide Hydrolases; Plasma; Transfusion Reaction

Sodium citrate is almost always used to anticoagulate blood for the preparation of washed platelet suspensions. Several adverse effects of citrate on platelet functional responses have been reported. We investigated the extent of activation of platelets and plasmatic coagulation during the preparation of washed platelets from human blood to which no anticoagulant was added. Washed platelets from native blood (PNB) were prepared by passing freshly-drawn human blood rapidly through a mixed Sephadex G-25/G-50 column to remove divalent cations. Gel-filtered blood (GFB), diluted by the elution medium containing 0.35% albumin and 2U/ml apyrase, was obtained within 5 minutes of venepuncture. Using CaCl2, the GFB was found to contain a mean of 1.65 X 10 mM calcium. Fibrinopeptide A measurements indicated less activation of plasmatic coagulation in GFB than in citrated blood. Measurements of beta-thromboglobulin and platelet factor 4 during the various stages of the preparation of PNB showed no platelet activation. No platelet aggregates, as measured by the platelet aggregate ratio method, were observed in GFB. Transmission electron microscopy showed intact discoid platelets similar to those in platelet-rich plasma. The reduced activation of platelets and of plasmatic coagulation was due to the more effective removal of calcium ions from the blood by gel filtration than chelation by citrate. PNB may thus provide a model for studying the requirements for calcium in platelet function in the absence of any citrate-platelet interactions.

Topics: Anticoagulants; beta-Thromboglobulin; Blood Platelets; Calcium; Cations, Divalent; Cell Separation; Chromatography, Gel; Citrates; Fibrinopeptide A; Hemolysis; Humans; Microscopy, Electron; Platelet Aggregation; Platelet Factor 4

An in vitro perfusion system was used to study the platelet reactivity of the following vascular graft materials when tested with human blood: expanded polytetrafluoroethylene (ePTFE), crimped Dacron Bionit (DB) and preclotted Dacron Bionit (DB/PC). These materials were simultaneously compared to silicone rubber (SR) using an identical perfusion circuit with the same donor's blood. All vascular graft materials tested in this in vitro perfusion system caused some degree of platelet activation as shown by a decrease in platelet count, an increase in platelet factor 3 activity, elevation of plasma levels of both platelet factor 4 and beta-thromboglobulin and decreased platelet aggregability. The observed platelet activation was striking for Dacron and especially preclotted Dacron, with ePTFE showing low levels of platelet activation. Platelet activation by Dacron was initially rapid and then levelled off, whereas the platelet activation with preclotted Dacron began more slowly, but reached much greater levels after three hours of in vitro perfusion.

Topics: beta-Thromboglobulin; Blood Coagulation; Blood Platelets; Blood Vessel Prosthesis; Fibrinogen; Fibrinopeptide A; Hemolysis; Humans; In Vitro Techniques; Platelet Aggregation; Platelet Factor 3; Platelet Factor 4; Polyethylene Terephthalates; Polytetrafluoroethylene; Silicones; Surface Properties