Ann Thorac Surg 2003;76:1234-1239
© 2003 The Society of Thoracic Surgeons
Original article: cardiovascular
ONO-6818, a novel, potent neutrophil elastase inhibitor, reduces inflammatory mediators during simulated extracorporeal circulation
Yukihiro Yoshimura, MDa,
Yuji Hiramatsu, MD, PhDa*,
Yukio Sato, MD, PhDa,
Satoshi Homma, MD, PhDa,
Yoshiharu Enomoto, MDb,
Tomoaki Jikuya, MD, PhDa,
Yuzuru Sakakibara, MD, PhDa
a Department of Cardiovascular Surgery, Institute of Clinical Medicine, University of Tsukuba, Tsukuba, Japan
b The Harrison Department of Research Laboratories, University of Pennsylvania, Philadelphia, Pennsylvania, USA
* Address reprint requests to Dr Hiramatsu, Department of Cardiovascular Surgery, University of Tsukuba, 1-1-1 Tennodai, Tsukuba 305-8575, Japan.
e-mail: yuji3{at}md.tsukuba.ac.jp
Presented at the Poster Session of the Thirty-ninth Annual Meeting of The Society of Thoracic Surgeons, San Diego, CA, Jan 31–Feb 2, 2003.
 |
Abstract
|
|---|
BACKGROUND: Among the serine proteases, neutrophil elastase is a powerful cytotoxic enzyme and plays a pivotal role in the inflammatory response associated with cardiopulmonary bypass. This study assesses the effects of the specific inhibition of neutrophil elastase by a novel, potent, low-molecular-weight neutrophil elastase inhibitor, ONO-6818. We hypothesized that ONO-6818 reduces inflammatory mediators and modulates adhesion molecules and the deformability of neutrophils during simulated extracorporeal circulation.
METHODS: Simulated extracorporeal circulation was established by recirculating fresh heparinized (3.75 U/mL) human blood for 120 minutes in a membrane oxygenator and a roller pump with and without 1.0 µmol/L of ONO-6818 (n = 9 for control group, n = 7 for ONO-6818 group). The neutrophil adhesion molecules, CD11b and L-selectin, and the cytoplasmic F-actin of neutrophils were measured by flow cytometry. Neutrophil deformability was evaluated using simulated silicon microcapillaries. Neutrophil elastase, interleukin 8, and C5b-9 were measured using enzyme immunoassay.
RESULTS: Neutrophil elastase levels were significantly lower in the ONO-6818 group. ONO-6818 significantly reduced interleukin 8 and C5b-9 production. ONO-6818 did not modulate changes of CD11b and L-selectin during recirculation. Cytoplasmic F-actin content and changes of neutrophil deformability did not significantly differ between the groups.
CONCLUSIONS: Inhibition of neutrophil elastase activity with ONO-6818 reduces further interleukin 8 production and the formation of the complement membrane attack complex, and this results in a reduction of neutrophil elastase levels during simulated extracorporeal circulation. This study suggests that specific neutrophil elastase inhibition with ONO-6818 is a feasible therapeutic option to attenuate the exaggerated inflammatory response associated with cardiopulmonary bypass.
|
Introduction
|
|---|
Cardiopulmonary bypass (CPB) is known to induce a whole body inflammatory reaction that is responsible for significant morbidity and mortality in cardiac surgery. During CPB, activated blood components are thought to be intertwined and form a vicious network, which contributes to the inflammatory response. Activated neutrophils play a central role in this circle of the inflammatory cascade, which involves a contact system, a complement system, cytokines, most of the blood cells, and endothelium. Activated neutrophils induce tissue injury by the release of toxic oxygen species and granule contents including neutrophil elastase. Among a number of serine proteases, neutrophil elastase is one of the most powerful cytotoxic enzymes because of its biologic effects [1]. Neutrophil elastase degrades connective tissue components such as elastin, proteoglycan, fibronectin, and collagen [2], and therefore may cause severe tissue damage and multiple organ dysfunction.
ONO-6818, a low-molecular-weight compound (C23H28N6O4; molecular weight, 452.51), is a specific, reversible, nonpeptide neutrophil elastase inhibitor. The inhibitory activity of ONO-6818 (Ki value) against human neutrophil elastase is 12.16 nmol/L. The half-life of the drug is approximately 2 to 3 hours. This agent competitively inhibits neutrophil elastase in humans, rats, and hamsters, but does not inhibit other proteases such as pancreatic elastase, trypsin, proteinase 3, cathepsin G, or matrix metalloproteinases [3]. It has been reported that ONO-6818 inhibits acute lung injury induced by neutrophil elastase in rats, minimizing lung hemorrhage and the accumulation of neutrophils in the lung [4].
This study assesses the effects of the specific inhibition of neutrophil elastase by ONO-6818 in a simulated extracorporeal circulation (SECC) model. We measured leukocyte and neutrophil counts, neutrophil adhesion molecules (CD11b and L-selectin), cytoplasmic F-actin content of neutrophils, neutrophil deformability determined by whole-blood transit time through simulated microcapillaries, plasma neutrophil elastase levels, interleukin 8 (IL-8), and the terminal complement complex C5b-9. We hypothesized that neutrophil elastase inhibition by ONO-6818 reduces inflammatory mediators and modulates adhesion molecules and the deformability of neutrophils during SECC.
|
Material and methods
|
|---|
Simulated extracorporeal circulation involved a spiral coil membrane oxygenator (model 60EC, surface area, 0.6 m2; MERA, Inc, Tokyo, Japan), a polyvinyl chloride venous reservoir bag (MERA), silastic tubing (1/4- and 3/8-inch inner diameter), polycarbonate connectors, and a barely occlusive roller head pump (model MS-033; MERA). Each circuit was primed with 250 mL of fresh human blood without dilution. Blood was obtained from healthy, fasting volunteers, who abstained from all medication for at least 2 weeks before donation. One donor was used for each individual bypass. Written informed consent was obtained from donors, and the protocol was approved by the Institutional Review Board of the University of Tsukuba. In the control (standard heparin) group (n = 9), blood was drawn directly into a reservoir bag containing standard heparin (3.75 U/mL) and dextrose (2.25 mg/mL). In the ONO-6818 group (n = 7), the reservoir bag contained heparin (3.75 U/mL), dextrose (2.25 mg/mL), and ONO-6818 (final concentration of 1.0 µmol/L blood). ONO-6818 was a generous gift from Ono Pharmaceutical Co, Osaka, Japan. Each individual circuit was used only once and then discarded. Blood was recirculated for 120 minutes at 400 mL/min with the blood temperature maintained at 37°C by immersing the reservoir bag in a constant-temperature shaking water bath. The oxygenator was ventilated with 95% oxygen–5% carbon dioxide at a rate of 1.0 L/min. Preliminary experiments confirmed that the pH of the circulating blood was maintained from 7.3 to 7.5 and that the activated clotting time was more than 500 seconds throughout the experiment. The circuit pressure was not measured or controlled.
Blood samples were obtained for analysis from each donor before any anticoagulant was introduced (donor sample), from the reservoir bag before beginning recirculation (0 minutes), and at 30, 60, and 120 minutes of recirculation. Additionally, a standing control sample (3.75 U/mL heparin and 2.25 mg/mL dextrose with or without ONO-6818) was collected from the reservoir bag and incubated for 120 minutes at 37°C. Blood samples for analysis were obtained with either 3.8% acid-citrate-dextrose (for CD11b, L-selectin, F-actin, and microchannel analysis, 9:1 by volume) or 1.0% ethylenediaminetetraacetic acid (EDTA-2Na; for neutrophil elastase, IL-8, and C5b-9). Blood collected with 1.0% EDTA-2Na was centrifuged immediately for 15 minutes at 2,000 g at 4°C. The plasma was then divided into aliquots and stored at -80°C for subsequent measurements. Samples for cell counts were collected in EDTA-2Na tubes (3.0 mg of EDTA-2Na per 2.0 mL of blood). The plasma concentration of ONO-6818 was not measured in this study.
Blood cell counts
Blood cell counts were performed using a counter (T-660; Coulter Electronics, Hialeah, FL), and differential white cell counts were made on Wright's stained blood smears by an experienced independent observer. Leukocyte and neutrophil counts were expressed as a percentage of the donor values.
Neutrophil elastase, interleukin-8, and C5b-9 assay
Plasma neutrophil elastase in a complex with 
1-protease inhibitor was determined by an activating immunization method (Merck Diagnostica, Darmstadt, Germany) with an automated homogeneous enzyme immunoassay. Enzyme-linked immunoassay for IL-8 was performed using commercially available kits (Pierce Endogen, Rockford, IL). Terminal complement complex C5b-9 was measured by an enzyme-linked immunosorbent assay (Quidel, San Diego, CA). The sensitivity limit of each assay undertaken was as follows: neutrophil elastase, 25 ng/mL; IL-8, 2 pg/mL; C5b-9, 16 ng/mL.
Adhesion molecules assay
Changes in the surface expression of L-selectin and CD11b of neutrophils were measured using flow cytometry as previously described [5]. One hundred microliters of whole blood samples were incubated for 30 minutes with 2 mg/mL of fluorescein isothiocyanate (FITC)–conjugated CD62L antibody (Pharmingen, San Diego, CA) and 1 mg/mL of phycoerythrin-conjugated mouse monoclonal anti-human CD11b antibody (DAKO Laboratories, Copenhagen, Denmark) at 4°C. Identical samples were incubated with FITC-conjugated mouse immunoglobin G (DAKO Laboratories) and phycoerythrin-conjugated mouse immunoglobin G2a (DAKO Laboratories) for negative control. The erythrocytes were lysed with Immuno-lyse, and leukocytes were fixed with Immuno-fix (Coulter Clone, Hialeah, FL). Neutrophils were identified using the typical forward and side-scatter pattern, and the expression of L-selectin and CD11b was measured as the mean fluorescent intensity of 5,000 cells. L-Selectin and CD11b changes were expressed as the percentage changes compared to the donor value.
F-actin content assay
A 50-µL sample was fixed with formaldehyde, and the cells were permeabilized using IntraPrep (Immunotech Coulter, Marseilles, France). Neutrophils were stained for 30 minutes at 37°C with 1 U of BODIPY FL phallacidin (Molecular Probes Inc, Eugene, OR). Cells were washed with phosphate-buffered saline solution, and the F-actin content was measured using a flow cytometer (FACS Calibur, Becton Dickinson, Franklin Lakes, NJ) as previously described [5]. The change in F-actin content was expressed as the percentage change from the donor value.
Neutrophil deformability assay
The transit time of the whole blood through the microchannel array was measured as a surrogate marker of neutrophil deformability. The detailed procedures and apparatus (Microchannel Array Flow Analyzer, MC-FAN, type KH-2; Hitachi Haramachi Electronics, Co, Hitachi, Japan) for this analysis have been previously described [6, 7]. In short, microgrooves formed in the surface of a single crystal silicon substrate were converted to leakproof microchannels by tightly covering them with an optical flat glass plate. The microgrooves in the silicon-made microchannel chip (Bloody-3S, 2600 channels; width, 6 µm; depth, 4.5 µm; length, 10 µm; Hitachi Haramachi Electronics), which are close to the size of capillaries, were prefilled with saline. Whole blood samples collected with 3.8% acid-citrate-dextrose were diluted with phosphate-buffered saline solution (1:1 by volume), and the suspension was made to flow through the microchannels under a pressure difference of 10 cm H2O. The transit time for each 100-µL suspension was determined to assess the filterability of the whole blood. These measurements were performed immediately after blood sampling, keeping the room temperature at 20° to 25°C. Results were expressed as a percentage of the transit time of the donor samples.
Statistical analysis
All values are expressed as mean ± standard error of the mean. One-way analysis of variance as compared with the donor value was used for within group comparison. Comparison of two groups as a function of time was performed by two-way analysis of variance with repeated measures. Data were further compared by the use of the Bonferroni test if analysis of variance was significant (p < 0.05).
|
Results
|
|---|
Changes in measured blood and plasma constituents and microchannel transit times during experiments are shown in Table 1.
The hematocrit value (data not shown) did not change significantly in either group throughout the recirculation. By 120 minutes of recirculation, leukocyte and neutrophil counts significantly decreased to approximately 86% to 92% of the donor value in both the control and the ONO-6818 groups, but there were no significant differences between the groups with time.
Plasma neutrophil elastase levels showed a significant 10-fold increase at 120 minutes of recirculation (p < 0.0001) in the control group. ONO-6818 significantly reduced this increase of neutrophil elastase. There were significant differences between the two groups at 30, 60, and 120 minutes of recirculation (Fig 1; p = 0.036, p = 0.027, and p = 0.021, respectively).
Plasma IL-8 levels increased significantly from 7.5 ± 3.7 to 175.0 ± 47.2 pg/mL by 120 minutes of recirculation (p = 0.0053) in the control group, whereas the ONO-6818 group showed a significant but smaller increase of IL-8 levels from 6.8 ± 1.6 to 36.1 ± 7.4 pg/mL by 120 minutes (p = 0.0023). Significant differences were noted between the two groups at 60 and 120 minutes of recirculation (Fig 2;
p = 0.0073 and p = 0.0093, respectively).
Complement C5b-9 levels increased significantly from 36.1 ± 7.4 to 1142.7 ± 135.0 ng/mL by 120 minutes of recirculation (p < 0.0001) in the control group. Increase of C5b-9 levels was attenuated in the ONO-6818 group. There were significant differences between the two groups at 120 minutes (Fig 3;
p = 0.044).
Expression of CD11b on the neutrophil surface increased significantly in both groups, reaching its highest values at 120 minutes of recirculation. No statistically significant differences were observed between the groups (Fig 4).
Expression of L-selectin on the neutrophil surface consistently decreased after 30 minutes of recirculation. There were no significant differences between the groups (Fig 5).
The cytoplasmic F-actin content of the neutrophils significantly increased to 161.0% ± 27.9% at 120 minutes (p = 0.040) in the control group. At other times, F-actin content did not reach statistical significance, with large standard errors. F-Actin content did not show any significant differences between groups.
The microchannel transit time of the blood increased significantly during recirculation in both groups: 169.4% ± 23.1% of the donor value in the control group and 169.9% ± 29.5% in the ONO-6818 group at 120 minutes (p = 0.013 and p = 0.045, respectively). However, there were no significant differences between the two groups.
|
Comment
|
|---|
On the basis of preliminary in vitro studies in human plasma that indicate that ONO-6818 inhibits the activity of neutrophil elastase in a dose-dependent fashion at doses of 0.1 to 10.0 µmol/L (50% inhibitory concentration of 0.403 µmol/L) at 37°C (unpublished data), we decided to use an initial blood concentration of 1.0 µmol/L in this first trial of the drug in SECC. The preliminary studies also demonstrate that ONO-6818 does not affect release of neutrophil elastase from neutrophils. Hence, reduction of plasma neutrophil elastase levels in the ONO-6818 group observed in this study is not thought to be the direct effect of this agent. In addition to tissue damage, neutrophil elastase promotes chemotaxis activation of neutrophils and expression of neutrophil adhesion molecules [8]. Neutrophil elastase also has the important biologic activity of inducing IL-8 [9], which can again stimulate the degranulation of neutrophils. In the present study, inhibition of neutrophil elastase activity with ONO-6818 led to reduction of IL-8, and probably prevented further neutrophil degranulation by IL-8 or neutrophil elastase itself. Thus, it could have resulted in reduction of neutrophil elastase levels.
Interleukin 8 is produced predominantly by activated neutrophils, monocytes, macrophages, and T cells. The elevation of plasma IL-8 levels has been shown during and after clinical CPB [10, 11], and is associated with cardiac and pulmonary dysfunction after CPB [12]. The delayed increase of IL-8 at 120 minutes of SECC observed in our study is consistent with previous in vivo [10, 11] and in vitro [13] studies. The effects of IL-8 on human neutrophils are not only chemotaxis and release of storage enzymes such as neutrophil elastase but also expression of surface adhesion molecules and production of reactive oxygen metabolites [14]. Hence, the fact that ONO-6818 attenuates IL-8 production means the drug possibly reduces neutrophil-mediated tissue injury associated with CPB.
Complement is activated by both classic and alternative pathways during CPB [15]. Complement activation generates the anaphylatoxins C3a and C5a and membrane attack complex C5b-9. Each of these substances has been implicated as an important cause of the inflammatory response [16]. C5b-9, formed as a byproduct of the pathway and released into the plasma, is relatively stable and a good marker of activation of the terminal complex pathway [17]. In previous studies, nafamostat mesilate completely attenuated neutrophil elastase release but failed to inhibit overall complement activity as judged by C5b-9 [18]. On the other hand, anti-factor D monoclonal antibody 166-32 inhibited the alternative complement cascade and reduced the production of C3a, C5b-9, IL-8, and neutrophil elastase and CD11b expression during SECC [13]. In a baboon CPB model, anti-factor D 166-32 reduced C3a, C5b-9, and IL-6 production and CD11b expression [19]. It has been suggested that neutrophil elastase is an important mediator in complement-mediated acute lung injury [20]. In the present study, we expected IL-8 reduction by neutrophil elastase inhibition, but the attenuation of C5b-9 in the ONO-6818 group was not expected with certainty. Our findings, as well as the results of previous studies, suggest that neutrophil elastase activity may have close interactions with complement activation and inflammatory cytokines inside the huge network of the inflammatory cascade.
Downregulation of L-selectin and upregulation of CD11b during SECC have been proven to be good markers of neutrophil activation [5, 6, 10, 21, 22]. In response to stimulation by chemotactic factors, there is a rapid and transient increase in F-actin assembly from the G-actin pool [23]. Changes in the neutrophil cytoskeleton with F-actin assembly at the cell periphery are thought to be responsible for cell deformability [23]. The decrease of deformability of neutrophils contributes to neutrophil sequestration in organ capillaries, allowing close proximity and adhesion of neutrophils to endothelial cells. Because neutrophil sequestration in organ capillaries is one of the initiating events in the development of the inflammatory response and multiple organ injuries [24], F-actin content and expression of adhesion molecules of neutrophils are important indicators for the inflammatory response. We have previously reported that SECC causes an increase of F-actin and a loss of neutrophil deformability estimated by whole-blood transit time through simulated microcapillaries, as well as an increase of CD11b and a decrease of L-selectin [6]. We have also reported that phosphodiesterase type 4 inhibitor rolipram attenuates expression of CD11b and F-actin and preserves neutrophil deformability in SECC [5].
In the present study, however, ONO-6818 did not modulate expression of adhesion molecules, cytoplasmic F-actin content, or the change of neutrophil deformability. Regarding F-actin content, there were relatively large standard errors with time in both groups as found in our previous studies [5, 6], which makes interpretation of the drug's effect on this cytoplasmic marker uncertain. It is unclear why ONO-6818 did not affect these adhesion molecules or cell deformability markers, despite significant attenuation of the plasma inflammatory mediators, neutrophil elastase, IL-8, and C5b-9 by the drug. The lack of endothelial cells, which have many molecules and function against neutrophil adhesion, could be the main reason. This may be a limitation of the current study on the nonendothelial SECC model, which mimics pediatric CPB systems with a small membrane oxygenator and small-diameter tubing. In the modulation of adhesion molecules and the cytoskeleton and the deformability of neutrophils, neutrophil elastase inhibitors may potentially have positive effects in the presence of endothelial cells, as has been demonstrated in a recent study which shows that neutrophil elastase enhances adhesion of neutrophils to endothelial cells [25]. Although significant limitations exist, a similar normothermic SECC system has been continuously used for many years to screen potential anticoagulants and protease inhibitors in a number of previous studies [5, 18, 26]. The advantage of the SECC model is that this totally in vitro approach can avoid loss of activated blood cells or markers from the circuit, and also avoids new cells being recruited from the bone marrow. It is a screen and is not intended to be a substitute for animal studies, which are not easy because most human antibodies to inflammatory markers do not cross-react with nonprimate laboratory animals. Further investigation of ONO-6818 in an in vivo primate CPB system would be necessary to reveal the entire effects of this promising drug on the inflammatory network in the presence of the endothelial system.
In conclusion, inhibition of neutrophil elastase with ONO-6818 reduces IL-8 production and formation of the terminal complement complex during SECC, but does not modulate neutrophil adhesion molecules, F-actin content, or neutrophil deformability. Neutrophil elastase inhibition with ONO-6818 is a feasible therapeutic strategy to prevent an exaggerated inflammatory response in CPB.
|
Acknowledgments
|
|---|
The authors thank Shoko Sato for her excellent technical support, Avi Landau for the language direction, Shiro Hinotsu, PhD, for statistical assistance, and Ono Pharmaceutical Co, Osaka, Japan, for the drugs and support for the study. A portion of the perfusion materials were kindly provided by MERA, Inc, Tokyo, Japan.
|
References
|
|---|
- Weissmann G., Smolen J.E., Korchak H.M. Release of inflammatory mediators from stimulated neutrophils. N Engl J Med 1980;303:27-34.[Medline]
- Havemann K., Gramse M. Physiology and pathophysiology of neutral proteinases of human granulocytes. Adv Exp Med Biol 1984;167:1-20.
- Ohmoto K., Okuma M., Yamamoto T., et al. Design and synthesis of new orally active inhibitors of human neutrophil elastase. Bioorg Med Chem 2001;9:1307-1323.[Medline]
- Kuraki T., Ishibashi M., Takayama M., Shiraishi M., Yoshida M. A novel oral neutrophil elastase inhibitor (ONO-6818) inhibits human neutrophil elastase-induced emphysema in rats. Am J Respir Crit Care Med 2002;166:496-500.[Abstract/Free Full Text]
- Sato Y., Hiramatsu Y., Homma S., Sato S., Onizuka M., Sakakibara Y. Phosphodiesterase type-4 inhibition of activated polymorphonuclear leukocytes in a simulated extracorporeal circulation model. J Thorac Cardiovasc Surg 2003;125:172-177.[Abstract/Free Full Text]
- Yoshimura Y., Hiramatsu Y., Sato Y., et al. Activated neutrophils and platelet microaggregates impede blood filterability through microchannels during simulated extracorporeal circulation. Ann Thorac Surg 2003;75:1254-1260.[Abstract/Free Full Text]
- Kikuchi Y. Effect of leukocytes and platelets on blood flow through a parallel array of microchannels: micro- and macroflow relation and rheological measures of leukocyte and platelet activities. Microvasc Res 1995;50:288-300.[Medline]
- Yamaguchi Y., Matsumura F., Wang F.S., et al. Neutrophil elastase enhances intercellular adhesion molecule-1 expression. Transplantation 1998;65:1622-1628.[Medline]
- Padilla M.L., Galicki N.I., Kleinerman J., Orlowski M., Lesser M. High cathepsin B activity in alveolar macrophages occurs with elastase-induced emphysema but not with bleomycin-induced pulmonary fibrosis in hamsters. Am J Pathol 1988;131:92-101.[Abstract]
- Finn A., Morgan B.P., Rebuck N., et al. Effects of inhibition of complement activation using recombinant soluble complement receptor 1 on neutrophil CD11b/CD18 and L-selectin expression and release of interleukin-8 and elastase in simulated cardiopulmonary bypass. J Thorac Cardiovasc Surg 1996;111:451-459.[Abstract/Free Full Text]
- Ashraf S., Tian Y., Cowan D., et al. "Low-dose" aprotinin modifies hemostasis but not proinflammatory cytokine release. Ann Thorac Surg 1997;63:68-73.[Abstract/Free Full Text]
- Hennein H., Rodriguez L., Ebba H., et al. Cytokine production associated with myocardial ischemia following uncomplicated coronary artery bypass grafting. Surg Forum 1992;43:223-225.
- Fung M., Loubser P.G., Undar A., et al. Inhibition of complement, neutrophil, and platelet activation by an anti-factor D monoclonal antibody in simulated cardiopulmonary bypass circuits. J Thorac Cardiovasc Surg 2001;122:113-122.[Abstract/Free Full Text]
- Baggiolini M., Walz A., Kunkel S.L. Neutrophil-activating peptide-1/interleukin 8, a novel cytokine that activates neutrophils. J Clin Invest 1989;84:1045-1049.
- Edmunds L.H., Jr Inflammatory response to cardiopulmonary bypass. Ann Thorac Surg 1998;66(Suppl):S12-28.[Abstract/Free Full Text]
- Kirklin J.K., Westaby S., Blackstone E.H., Kirklin J.W., Chenoweth D.E., Pacifico A.D. Complement and the damaging effects of cardiopulmonary bypass. J Thorac Cardiovasc Surg 1983;86:845-857.[Abstract]
- Videm V., Svennevig J.L., Fosse E., Semb G., Osterud A., Mollnes T.E. Reduced complement activation with heparin-coated oxygenator and tubings in coronary bypass operations. J Thorac Cardiovasc Surg 1992;103:806-813.[Abstract]
- Sundaram S., Gikakis N., Hack C.E., et al. Nafamostat mesilate, a broad spectrum protease inhibitor, modulates platelet, neutrophil and contact activation in simulated extracorporeal circulation. Thromb Haemost 1996;75:76-82.[Medline]
- Undar A., Eichstaedt H.C., Clubb F.J., Jr, et al. Novel anti-factor D monoclonal antibody inhibits complement and leukocyte activation in a baboon model of cardiopulmonary bypass. Ann Thorac Surg 2002;74:355-362.[Abstract/Free Full Text]
- Hagio T., Nakao S., Matsuoka H., Matsumoto S., Kawabata K., Ohno H. Inhibition of neutrophil elastase activity attenuates complement-mediated lung injury in the hamster. Eur J Pharmacol 2001;426:131-138.[Medline]
- Le Deist F., Menasche P., Kucharski C., Bel A., Piwnica A., Bloch G. Hypothermia during cardiopulmonary bypass delays but does not prevent neutrophil-endothelial cell adhesion: a clinical study. Circulation 1995;92(Suppl II):II-354-358.
- Galinanes M., Watson C., Trivedi U., Chambers D.J., Young C.P., Venn G.E. Differential patterns of neutrophil adhesion molecules during cardiopulmonary bypass in humans. Circulation 1996;94(Suppl II):II-364-369.
- Frank R.S. Time-dependent alterations in the deformability of human neutrophils in response to chemotactic activation. Blood 1990;76:2606-2612.[Abstract/Free Full Text]
- Toren M., Goffinet J.A., Kaplow L.S. Pulmonary bed sequestration of neutrophils during hemodialysis. Blood 1970;36:337-340.[Abstract/Free Full Text]
- Nozawa F., Hirota M., Okabe A., et al. Elastase activity enhances the adhesion of neutrophil and cancer cells to vascular endothelial cells. J Surg Res 2000;94:153-158.[Medline]
- Gikakis N., Khan M.M.H., Hiramatsu Y., et al. Effect of factor Xa inhibitors on thrombin formation and complement and neutrophil activation during in vitro extracorporeal circulation. Circulation 1996;94(Suppl II):II-341-346.
This article has been cited by other articles:
|
|
|
|
 
K. Matsuzaki, Y. Hiramatsu, S. Homma, S. Sato, O. Shigeta, and Y. Sakakibara
Sivelestat Reduces Inflammatory Mediators and Preserves Neutrophil Deformability During Simulated Extracorporeal Circulation
Ann. Thorac. Surg.,
August 1, 2005;
80(2):
611 - 617.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
Y. Hiramatsu, S. Homma, Y. Sato, S. Sato, K. Matsuzaki, O. Shigeta, and Y. Sakakibara
Nafamostat Preserves Neutrophil Deformability and Reduces Microaggregate Formation During Simulated Extracorporeal Circulation
Ann. Thorac. Surg.,
April 1, 2005;
79(4):
1326 - 1332.
[Abstract]
[Full Text]
[PDF]
|
|
|
Top
Abstract
Introduction
p < 0.05, 
p < 0.001 by one-way analysis of variance as compared with the donor value. (SC = standing control.)
p < 0.01 by one-way analysis of variance as compared with the donor value. (SC = standing control.)
