Reduction of complement activation during bypass by prime manipulation

Complement activation is believed to be of importance in the development of complications arising after cardiopulmonary bypass. The effect on complement activation of priming the extracorporeal circuit with crystalloid alone, crystalloid plus albumin, or crystalloid plus the plasma expander polygeline was assessed in 36 patients undergoing coronary artery operations with cardiopulmonary bypass using a bubble oxygenator. Activation of the alternative and common complement pathways was monitored before, during, and after the bypass period by measuring concentrations of factor B and its fragment Ba and C3 and its fragment C3d. Complement activation occurred in all three groups of patients, with no difference between the crystalloid and crystalloid-albumin groups. In contrast, Ba fragment concentrations were persistently and significantly lower during and after bypass in the polygeline group, denoting reduced complement activation. C3d levels also showed a tendency to be lower in this group. Our results indicate that addition of polygeline to the priming solution reduces complement activation. Because complement activation is associated with morbidity after cardiopulmonary bypass, addition of polygeline to the priming solution may offer an inexpensive method of reducing morbidity after cardiopulmonary bypass.     

Different activation patterns in the plasma kallikrein-kinin and complement systems during coronary bypass surgery

Components of the plasma kallikrein-kinin and complement systems were determined in patients undergoing open heart surgery with cardiopulmonary bypass. Spontaneous kallikrein activity (KK), plasma prekallikrein (PKK), functional kallikrein inhibition capacity (KKI), C3 activation products (C3-act), and the terminal complement complex (TCC) were measured. A marked, transitory increase in KK and a decrease in PKK were found prior to cardiopulmonary bypass just after heparin injection. An additional decline in PKK and KKI during bypass with a return to near control levels in the postoperative period was observed. C3-act increased in all patients during bypass, reaching a peak value at wound closure. The TCC concentration also increased significantly during cardiopulmonary bypass, returned to control levels in the early postoperative period, and then increased again in the late postoperative period. It is concluded that activation of the kallikrein-kinin system started after injection of heparin, prior to cardiopulmonary bypass. Activation of both the initial and the terminal complement cascade, however, started only after onset of cardiopulmonary bypass.     

Complement activation during major operations with or without cardiopulmonary bypass

Plasma concentrations of the complement products C3dg and the terminal complement complex, as well as the number of granulocytes (polymorphonuclear neutrophils), were assessed in patients undergoing aorta-coronary bypass with extracorporeal circulation, abdominal aneurysmectomy with implantation of an aortic graft, or thoracotomy without the introduction of synthetic material into the circulation. The concentration of terminal complement complex increased significantly only in the group undergoing extracorporeal circulation, with a corresponding drop in the number of granulocytes. In contrast, the C3dg concentration increased during both extracorporeal circulation and abdominal aneurysmectomy, which indicates that other factors than extracorporeal circulation may affect C3 activation during major operations. In the thoracotomy group, where the most pronounced increase in granulocytes was found, no complement activation was recorded. It is concluded that extracorporeal circulation activates the terminal pathway of complement and that assays detecting activation of both the initial and the terminal parts should be included when the pathophysiology of complement is examined during major operations and extracorporeal circulation.     

Complement activation in extracorporeal circulation. Comparison of nylon versus polyester bubble oxygenators

Complement activation during cardiopulmonary bypass is well known and may influence postoperative morbidity. As nylon can particularly induce complement activation, its influence was assessed by measuring total haemolytic complement and B, C3 and C4 factors, during cardiopulmonary bypass with bubble oxygenators for coronary surgery, comparing "nylon" circuits (20 patients, Bentley BOS 10) versus "polyester" circuits (19 patients, Shiley S 100 A). Complement activation began with induction of anaesthesia and surgical procedures, B, C3 and C4 levels falling significantly (respectively 15, 17 and 20% from baseline values). The alternative pathway was activated before the classical pathway. Complement activation continued during cardiopulmonary bypass, with no more consumption of complement factors (slight variations of about 0 to 3% of the levels found after anaesthetic induction and surgical procedures). No statistically significant difference appeared between the two groups. This suggested that nylon did not significantly increase complement activation during cardiopulmonary bypass. The bubble oxygenator material cannot therefore be considered as a criterion for choosing the type of equipment.     

Plasma fibronectin and complement in surgical patients

Plasma fibronectin and a fragment of the third component of complement (C3b) are both opsonins of the reticulo-endothelial system. Twenty patients undergoing uncomplicated gastro-intestinal surgery were studied. There was a significant reduction in plasma fibronectin and total haemolytic complement, but not C3, in the early postoperative period (days 1 to 3). In six surgical patients critically ill with severe sepsis both plasma fibronectin concentrations and C3 concentration were reduced significantly when compared with the early postoperative period of uncomplicated surgery. In addition, in the critically ill patients, direct evidence of complement activation was demonstrated. These studies show a transient reduction of humoral host defence mechanisms following surgery which may be permissive to bacteraemia and complement activation.     

PERIOPERATIVE CHANGES IN COMPLEMENT ASSOCIATED WITH CARDIOPULMONARY BYPASS

The total haemolytic complement (CH₅₀), the complement componentsC3 and C4, the complement breakdown product C3d, alternativepathway activation and transferrin, were measured before, duringand after cardiopulmonary bypass. As expected, CH₅₀o decreasedafter heparinization, remained low during bypass and decreasedfurther up to 8 h after bypass. C3 and C4 decreased significantlyduring bypass, continued to decrease for a further 8 h afterbypass (by 35% and 40% respectively) and thereafter increasedgradually up to 48 h. Although the depletions observed weresuggestive of complement activation, there were no demonstrableincreases in C3d, and in all patients the concentration of C3dremained within the normal range. Hence it was concluded thatcomplement depletions of this magnitude were unlikely to resultfrom complement activation. Non-specific changes in proteinconcentrations during bypass, as a result of dilution, redistributionor other unidentified factors, are more probable causes of theobserved reductions. The acute phase response to surgery maybe a factor in the subsequent increase in C3 and C4 which isseen 24 h after bypass. As transfenn concentrations in the plasmaare known to decrease during this response the observed decreasein transfcrrin concentration would support this view. ^*Present address: Department of Anaesthetics, The MiddlesexHospital, Mortimer Street, LondonWl.     

Time for new concepts about measurement of complement activation by cardiopulmonary bypass?

Fifty-one patients admitted for routine coronary bypass operations were randomized to cardiopulmonary bypass with a membrane oxygenator (Capiox) or a bubbler (Polystan or William Harvey). Complement activation was measured using enzyme immunoassays for concentrations of C3 activation products and the terminal complement complex. From 5.8 to 8.1 arbitrary units (AU)/mL (medians), the plasma concentrations of C3 activation products increased by 119.9 AU/mL (Capiox), 124.6 AU/mL (Polystan), and 79.5 AU/mL (William Harvey) to a peak at closure of the sternum (not significant when related to baseline concentrations). The increase in C3 activation products and baseline C3 activation were linearly correlated (R2 = 0.30; p less than 0.0001). From 5.5 to 6.1 AU/mL, the plasma terminal complement complex concentrations increased by 45.2 AU/mL (Capiox), 15.4 AU/mL (Polystan), and 17.4 AU/mL (William Harvey) to a peak before termination of cardiopulmonary bypass. Maximal terminal (C5-C9) activation was significantly higher in the membrane oxygenator group (p less than 0.0001) and showed no relationship to C3 activation. Measurement of C3 activation only gives no information about C5-C9 activation. At present, terminal complement complex quantitation is probably the best index of C5-C9 activation during cardiopulmonary bypass.     

Relationship between granulocyte elastase and C3a under protamine dosing in on-pump cardiac surgery.

OBJECTIVE: The complement cascade and granulocytes are activated in on-pump cardiac surgery. If activation of complement directly regulates granulocytes, granulocyte elastase (GEL) should increase significantly after protamine administration. We examined the effect of protamine on granulocytes by protamine administration and observation of the effect on GEL and C3a. METHODS: Thirty patients who underwent coronary artery bypass grafting were randomly assigned to two groups. In 15 patients, protamine was administered 5 min after the termination of cardiopulmonary bypass, and was administered 35 min after cardiopulmonary bypass in the other 15 patients. All patients were perfused with heparin-coated circuits and received 300 IU/kg heparin and 3 mg/kg protamine. GEL and C3a concentrations were measured at 7 time points. RESULTS: GEL concentrations increased significantly just before aortic declamping and did not increase significantly after protamine administration. C3a concentrations, however, did not increase during cardiopulmonary bypass and did increase significantly after protamine administration. CONCLUSIONS: This study indicates that GEL does not increase after protamine administration and that complement concentration does not directly affect GEL release.     

Early postoperative changes with different priming solutions in open-heart surgery

The consequences of substituting dextran 70 (Macrodex) for human plasma in the priming solution were studied during the first 48 hours after open-heart surgery. Twenty patients undergoing elective surgery for coronary artery disease were selected for the study. Clinical, haemodynamic, metabolic and roentgenologic parameters were monitored. No major differences were found between results with use of plasma or dextran (each in 10 patients). The lower pulmonary capillary wedge pressure in the dextran group may indicate better left ventricular performance. As human plasma is a limited resource and is a potential disease transmitter, dextran 70 may with advantage replace human plasma in the priming solution, and at much lower cost.     

Induction of acute-phase reactive substances during open-heart surgery and efficacy of ulinastatin. Inhibiting cytokines and postoperative organ injury.

OBJECTIVE: A systemic inflammatory response after open-heart surgery using cardiopulmonary bypass may be responsible for postoperative organ dysfunction. Ulinastatin, a protease inhibitor, plays an important role in host defense under periods of stress. METHODS: We studied the efficacy of ulinastatin on changes in acute-phase reactive substances during and after open-heart surgery. Patients undergoing open-heart surgery were divided into an ulinastatin group (Group U) and a control group (Group C). In Group U, we introduced 600,000 units of ulinastatin into a priming solution for cardiopulmonary bypass, 300,000 units into a cardiopulmonary bypass circuit at the removal of aortic cross-clamping, and 300,000 units a day for 5 days following surgery. RESULTS: Immediately after cardiopulmonary bypass, alpha 1-antitrypsin levels decreased significantly in both groups, and increased significantly on the second day after surgery. Ulinastatin levels decreased after cardiopulmonary bypass in Group C. Significantly high levels of ulinastatin were obtained in Group U. Interleukin-6, interleukin-8, and polymorphonuclear elastase were markedly induced, and high levels of plasma concentration continued for several days after surgery. At all sample points, these concentrations in Group U tended to be lower than those in Group C. A significantly positive correlation was seen between the maximum levels of interleukin-8 and polymorphonuclear elastase, but these cytokine and polymorphonuclear elastase levels did not correlate with parameters such as the duration of anesthesia, surgery, cardiopulmonary bypass, or aortic cross-clamping. CONCLUSIONS: Our study suggests that high-dose ulinastatin administration to maintain a sufficient concentration of circulating protease inhibitors may suppress overinduction of cytokines and polymorphonuclear elastase in open-heart surgery.     

Complement activation before, during and after cardiopulmonary bypass

Plasma levels of the complement parent molecules C3, C4, and factor B and their split products, C3d, C4d, and Ba were measured in 12 patients undergoing cardiopulmonary bypass for coronary artery surgery. Alternative and common complement pathway activation, demonstrated by statistically significant rising levels of Ba (P less than 0.05), and C3d (P less than 0.05) and by elevated Ba:B (P less than 0.05) and C3d:C3 (P less than 0.05) ratios were found before the institution of cardiopulmonary bypass but following heparin administration suggesting that heparin may itself initiate alternative pathway activation. In addition, significant depletion of parent complement components and elevation of split product concentrations was seen during bypass suggesting classical and alternate pathway activation (P less than 0.01). This study clarifies the pathways of complement activation during bypass and presents evidence that heparin administration may initially activate the complement cascade.     

Plasma fibronectin and complement activation in coronary bypass surgery

In patients with severe sepsis, plasma fibronectin concentrations are reduced and complement is activated. It is not known whether complement activation interferes with plasma fibronectin. Cardiopulmonary bypass is known to activate complement. We have therefore used this operation to study the effect of complement activation on plasma fibronectin in the absence of sepsis. After 15 min of bypass the percentage changes of plasma fibronectin and haematocrit were similar (65.9 +/- s.e.m. 4.8 and 67.0 per cent +/- s.e.m. 2.3, respectively), but the changes in C3 and C5 (58.4 +/- s.e.m. 4.8 per cent and 52.5 per cent +/- s.e.m. 2.1) were significantly greater than those of the haematocrit, indicating complement consumption. During the first 60 min of bypass there was a significant increase in the neutrophil count which is compatible with complement activation. It is unlikely that complement activation alone, in the absence of sepsis, contributes to the reduction of plasma fibronectin concentrations.     

Peri-operative changes in plasma C3a and C5a concentrations in infants

It is well known that complement activation plays a key role in the development of pulmonary insufficiency; it is also well known that cortisol suppresses complement activation. By measuring the plasma concentrations of C3a and C5a, we investigated peri-operative changes in the activation of the complement system in 18 infants undergoing elective abdominal surgery. We also measured plasma cortisol concentrations to assess the peri-operative relationship between complement activation and the stress produced by multiple peri-operative factors following two anaesthetic techniques. Eighteen infants ranging in age from 1 to 11 months were randomly divided into two groups according to the anaesthetic technique used: Group 1 consisted of nine infants in whom general anaesthesia was maintained with halothane and nitrous oxide (N₂O) in oxygen, while Group 2 consisted of nine infants in whom general anaesthesia was maintained with fentanyl and N₂O in oxygen. Plasma C3a and C5a concentrations were higher in the fentanyl group than in the halothane group during the peri-operative period. Plasma cortisol concentration, in contrast, was lower in the fentanyl group both during and after surgery. The post-operative clinical course showed no significant intergroup differences between the two groups throughout the study. These observations suggest that the difference in peri-operative complement activation between the halothane and the fentanyl groups may have been due, in part, to different peri-operative stress responses. However, further studies are required to elucidate the precise causative mechanisms and the clinical implications of complement activation in infants.     

Complement activation and lung permeability during cardiopulmonary bypass

Pulmonary dysfunction after cardiopulmonary bypass has been attributed to the damaging effects of complement activation on the lung. To further explore this phenomenon, we measured plasma levels of activated complement components (radioimmunoassay), assessed neutrophil n-formyl-methionyl-leucyl-phenylalanine (FMLP) receptor status (radioligand saturation binding assay), and quantified pulmonary epithelial permeability as radioaerosol lung clearance of technetium 99m-labeled diethylenetriamine pentaacetic acid in a series of 8 patients undergoing cardiopulmonary bypass. Significant elevations of plasma C3adesArg, C4adesArg, and C5adesArg levels were seen just after CPB, indicating activation of both the classic and alternate complement pathways. Neutrophil activation was evident as increased expression of neutrophil FMLP surface receptors after bypass. Despite the presence of complement and neutrophil activation, increased pulmonary epithelial permeability was not seen. These data support the hypothesis that complement and neutrophil activation during cardiopulmonary bypass is not associated with acute lung injury, at least not pulmonary epithelial injury. One can therefore infer that increased pulmonary epithelial permeability in patients at high risk for and experiencing sepsis-induced and trauma-induced adult respiratory distress syndrome may be due to factors other than complement and neutrophil activation.     

Superior biocompatibility of heparin-bonded circuits in pediatric cardiopulmonary bypass.

BACKGROUND: Heparin bonding of pediatric cardiopulmonary bypass circuits may decrease activation of blood compartments as inflammatory responses. We studied the biocompatibility of heparin-bonded circuits in infant cardiac surgery. METHODS: Twenty-four infants undergoing elective cardiac surgery were randomly assigned to either a nonheparin-bonded control circuit (n = 12) or a fully heparin-bonded circuit (n = 12) including membrane oxygenator, reservoir, and all tubing. Blood samples were used to identify differences in complement activation and cytokine release between groups during and after cardiopulmonary bypass. The postbypass oxygenation index was also compared. RESULTS: The C3 activation product in the heparin-bonded group was significantly lower during (p < 0.01) and just after (p < 0.05) cardiopulmonary bypass. No statistically significant difference in C4 activation products was observed. Lower interleukin-6 and tumor necrosis factor-alpha were found immediately after cardiopulmonary bypass (p < 0.05) and a higher mean postbypass oxygenation index was also seen (p < 0.05) in the heparin-bonded group. CONCLUSION: We found that a heparin-bonded cardiopulmonary bypass circuit reduced inflammatory response and improved oxygenation in pediatric cardiac surgery. These results suggest that the superior biocompatibility of the bonded circuit may reduce pulmonary complications.     

Heparin-coated cardiopulmonary bypass equipment. II. Mechanisms for reduced complement activation in vivo

OBJECTIVE: Our objective was to study mechanisms for reduced complement activation by heparin coating of cardiopulmonary bypass equipment in clinical heart surgery. METHODS: Adults undergoing elective coronary artery bypass grafting were randomized to cardiopulmonary bypass with Duraflo II heparin-coated (n = 15) or uncoated (n = 14) sets (Duraflo coating surface; Baxter International, Inc, Deerfield, Ill). Blood samples were analyzed with the use of enzyme immunoassays for C1rs-C1 inhibitor complexes and the activation products Bb, C4bc, C3bc, C5a-desArg, and the terminal complement complex. Data were compared by repeated-measures analysis of variance. RESULTS: C1 was activated during bypass, and increases in C1rs-C1 inhibitor complexes were larger with heparin coating (P =.03). C4bc increased after administration of protamine, without intergroup differences (P =.69). Bb (P =.22) and C5a-desArg (P =.13) tended to increase less with heparin coating. Formation of C3bc (P =.03) and the terminal complement complex (P <.01) was significantly reduced with heparin coating. C5a-desArg increased 2-fold during bypass, whereas the terminal complement complex increased 10- to 20-fold. Maximal terminal complement complex concentrations were significantly correlated to maximal Bb and C3bc (R = 0.6, P <.001), but not to C1rs-C1 inhibitor complexes or C4bc (R < 0.05, P >.8). CONCLUSIONS: C1 activation during bypass was increased by heparin coating, but further classical pathway activation was held in check until administration of protamine. Heparin coating significantly inhibited C3bc and terminal complement complex formation. Terminal complement complex concentrations were related to alternative pathway activation and may be useful for evaluation of differences in bypass circuitry. Increases and intergroup differences in terminal complement complex concentrations were much larger than those in C5a-desArg.     

Complement activation during cardiopulmonary bypass: mechanism and prevention

The mechanism of complement activation during cardiopulmonary bypass was studied for the prevention. In ten patients undergoing open-heart procedures, the serum levels of complement fractions (C3, C4, and C3 activator) were measured by a single radial immune diffusion method. In four of ten patients, the plasma levels of C3a, C4a, and C5a fractions were studied by the radioimmunoassay 2 antibodies method. The serum levels of C3, C4, and C3 activator decreased after cardiopulmonary bypass. The plasma levels of C3a, C4a, and C5a increased after bypass. The lower level of C3 activator shows that C3 activator was not excessively produced during cardiopulmonary bypass. Therefore it can be thought that much C4b2a from classical pathway as well as C3a over-production localizing extra-corporeal circuits and little inhibitors on alternative pathway resulted in increased complement activation. The prevention should be done from these etiologies.     

Induction of interleukin-1 production in patients undergoing cardiopulmonary bypass

Systemic reactions resembling inflammation occur in patients undergoing cardiopulmonary bypass. We now report that interleukin-1, an endogenous pyrogen and a key mediator of inflammation, is transiently and consistently generated in vivo by circulating monocytes within hours after cardiopulmonary bypass. Interleukin-1 production was assessed by measuring interleukin-1 functional activity and interleukin-1 beta antigen concentration in cell lysates from monocytes of patients during and after bypass. There was no increase in intracellular interleukin-1 activity during bypass and within the first hours after bypass, possibly because of a suppressive effect of hypothermia on interleukin-1 production, as documented in vitro. Maximal generation of interleukin-1 was observed 24 hours after extracorporeal circulation, concomitantly with the occurrence of a peak in body temperature. The amount of interleukin-1 generated at that time was linearly correlated with the increase in patients' body temperature. The peak in interleukin-1 production followed by 20 hours the peak in complement activation as assessed by determining C3a desArg and C5a desArg concentrations in patients' plasma. These results indicate that interleukin-1 may be involved in the pathogenesis of adverse systemic reactions associated with cardiopulmonary bypass.     

Neutrophil degranulation and complement activation during fetal cardiac bypass

BACKGROUND: Fetal cardiac bypass results in dysfunction of the fetoplacental unit (FPU) characterized by increased placental vascular resistance and respiratory acidosis. However the mechanisms of this dysfunction are not completely understood. To test the hypothesis that complement activation and neutrophil degranulation may contribute to the placental dysfunction associated with fetal bypass, we compared placental hemodynamics, complement activation, and neutrophil degranulation among fetuses exposed to cardiac bypass with a miniaturized bypass circuit including an in-line axial flow pump (Hemopump), fetuses undergoing bypass with a conventional roller pump circuit, and control fetuses that were similarly exposed but did not undergo bypass. METHODS: Twenty-six Western Cross sheep fetuses (median 122 days gestation) were randomly assigned to undergo cardiac bypass for 30 minutes with the Hemopump circuit (n = 8), to undergo bypass for 30 minutes with the conventional (roller pump) circuit (n = 10), or to undergo identical exposure and cannulation but not bypass (n = 8, controls). Blood samples were collected to measure white cell count and differential, and C3a and lactoferrin levels prior to bypass, at the end of bypass, and 1 and 2 hours after bypass. Hemodynamics and blood gases were also monitored. RESULTS: There was a fall in white cell count over time that continued after bypass in all groups; neutrophils and lymphocytes were affected similarly. C3a levels rose significantly from prebypass to postbypass in the roller pump group (p<0.0001) but not in either of the other groups. Lactoferrin levels rose significantly from start of bypass in both bypass groups (Hemopump p = 0.01; roller pump p<0.0001) but not in controls. The elevation in lactoferrin level coincided with worsening placental gas exchange and deteriorating cardiac function. CONCLUSIONS: Complement and neutrophil activation occurred with fetal cardiac bypass but only neutrophil activation mirrored the FPU and cardiac dysfunction, suggesting that products of neutrophil activation may be important contributing factors. Improved FPU function with a bypass circuit that has less extracorporeal surface and does not require a large priming volume may be due in part to a reduction in the magnitude of this inflammatory response.     

Neutrophil and macrophage activation and anaphylatoxin formation in orthotopic liver transplantation without the use of veno-venous bypass

Background. Activation of neutrophils and activation of complement may be an aetiologic factor behind circulatory insufficiency in association with reperfusion of the grafted liver.
Methods. Neutrophil and macrophage activation (determined as PMN elastase and neopterin release) and complement activation were evaluated in 15 consecutive patients undergoing orthotopic liver transplantation without the use of veno-venous bypass.
Results. The PMN elastase concentrations were increased at the end ot the anhepatic phase, 2, 5 and 30 min after start of reperfusion and 6 and 24 h postoperatively. There were significantly higher PMN elastase concentrations in patients with circulatory instability (postreperfusion syndrome) compared with those without postreperfusion syndrome. The neopterin concentration was increased 2 min after the start of reperfusion and remained elevated until 6 h postoperatively. The plasma complement C3a concentrations were increased at the end of the anhepatic phase and 2, 5 and 30 min after the start of reperfusion. The plasma C3a levels were higher in patients with postreperfusion syndrome compared to those without.
Conclusions. Activation of neutrophils and macrophages and of the complement cascade with the formation of biologically active substances may be one explanation for the circulatory instability often seen in patients undergoing orthotopic liver transplantation.