Ann Thorac Surg 2003;75:1254-1260
© 2003 The Society of Thoracic Surgeons
Original article: cardiovascular
Activated neutrophils and platelet microaggregates impede blood filterability through microchannels during simulated extracorporeal circulation
Yukihiro Yoshimura, MDa,
Yuji Hiramatsu, MD, PhDa*,
Yukio Sato, MD, PhDa,
Satoshi Homma, MD, PhDa,
Yoshiharu Enomoto, MDa,
Yuji Kikuchi, PhDb,
Yuzuru Sakakibara, MD, PhDa
a Department of Cardiovascular Surgery, Institute of Clinical Medicine, University of Tsukuba, Tsukuba, Japan
b National Food Research Institute, Tsukuba, Japan
Accepted for publication November 1, 2002.
* 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
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Abstract
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BACKGROUND: Neutrophil sequestration and platelet microaggregates in organ capillaries have been implicated in the inflammatory response associated with cardiopulmonary bypass. We examined the filterability of neutrophils and platelet microaggregates through silicon microchannels during simulated extracorporeal circulation. We hypothesize that blood contact with artificial surfaces over time decreases the ability of neutrophils, platelets, and their aggregates to pass through microchannels.
METHODS: Fresh human blood from donors (n = 9) was recirculated for 120 minutes in a simulated extracorporeal circuit. Blood samples were obtained from a donor at 0, 30, 60, and 120 minutes of recirculation. The microchannel transit time and the flow behavior of blood cells were evaluated by a silicon microchannel array flow analyzer. CD11b, L-selectin, and F-actin of neutrophils were measured by flow cytometry. Neutrophil and platelet counts and platelet aggregation to adenosine diphosphate were measured.
RESULTS: The microchannel transit time was prolonged during recirculation, reaching 185.9% ± 25.6% of baseline at 120 minutes. The video microscope showed that neutrophils and platelet microaggregates plugged the microchannels. CD11b, L-selectin, and F-actin levels changed significantly by 120 minutes. Platelet counts decreased and platelet aggregability was attenuated.
CONCLUSIONS: Simulated extracorporeal circulation caused a progressive loss in the ability of neutrophils, platelets, and their aggregates to pass through the microchannels independent of neutrophil adhesion molecule expression.
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Introduction
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The activation of blood elements mediates the principal complications associated with cardiopulmonary bypass (CPB) [1]. Neutrophils are the main target in the effort to control the whole-body inflammatory response associated with CPB [2]. The initiating event in the development of the inflammatory response and multiple organ injuries is neutrophil sequestration in the capillaries of organs [3]. Activated platelets form aggregates and microemboli as well as microaggregates with circulating monocytes and neutrophils [4, 5].
We examined the rheologic changes in neutrophils and platelets activated by CPB by using simulated microcapillaries. The microchannel filterability of whole blood was examined, and the deformability of activated neutrophils and platelets was visualized by a microchannel array flow analyzer during simulated extracorporeal circulation (SECC). The surface expression of neutrophil adhesion molecules (CD11b and L-selectin) and cytoplamic F-actin (one of the major stress fibers of neutrophils), plateletcounts, and the platelet aggregation response to adenosine diphosphate (ADP) were assessed to determine the correlation between neutrophil and platelet activation and the rheologic phenomenon. We hypothesized that blood contact with nonendothelial surfaces of SECC over time causes a progressive loss in the ability of neutrophils, platelets, and their aggregates to pass through the inorganic simulated microcapillaries.
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Material and methods
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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, Inc., Tokyo, Japan), silicone elastomer tubing (1/4- and 3/8-inch inner diameter; MERA, Inc., Tokyo, Japan), polycarbonate connectors, and a barely occlusive roller head pump (model MS-033; MERA, Inc., Tokyo, Japan). Each circuit was primed with 250 mL of fresh human blood obtained from healthy, fasting volunteers (n = 9). Donors abstained from all medication for at least 2 weeks before donation. One donor was used for each simulated bypass. Written informed consent was obtained from donors, and the protocol was approved by the Institutional Review Board of the University of Tsukuba.
Each circuit was used only once, then discarded. Blood was drawn directly into a reservoir bag containing standard heparin (3.75 U/mL) and dextrose (2.25 mg/mL). 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 experimental period.
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) was collected from the bag and incubated for 120 minutes in a shaking water bath at 37°C before processing. Blood samples for analysis were obtained with either 3.8% sodium citrate (for platelet aggregation) or 3.8% acid-citrate-dextrose (for Cd11b, L-selectin, F-actin, and microchannel analysis, 9:1 by volume). Samples for cell counts were collected in ethylenediamenetetra-acetic acid (EDTA)-2Na tubes (3.0 mg EDTA-2Na per 2.0 mL of blood).
Blood cell counts
Blood cell counts were performed using a cell counter (T-660; Coulter Electronics, Hialeah, FL), and differential white cell counts were made on Wright’s stain blood smears by an experienced independent observer.
Platelet counts and platelet aggregation
Blood used for platelet counts and aggregation was centrifuged at 150 x g for 10 minutes to prepare platelet-rich plasma and then at 15,000 x g to prepare platelet-poor plasma. Platelet counts of platelet-rich plasma were performed on a model Z1 Coulter counter (Coulter Electronics, Hialeah, FL). Platelet aggregation in response to ADP was assessed on an aggregometer (Model PAC-4S; NBS HEMA TRACER, Tokyo, Japan) with the use of 150,000 platelets/µL. Threshold doses of ADP were determined as the lowest doses of agonist needed to produce biphasic aggregation of at least 60.0% after 5 minutes in donor samples. Threshold doses of ADP were then used to measure the aggregation as a percentage of light transmittance in subsequent samples.
F-actin content assay
Fifty microliters of sample was fixed with formaldehyde, and the cells were permeabilized using IntraPrep permeabilization reagent (Immunotech; Coulter, Marseilles, France). Neutrophils were stained for 30 minutes in the dark at 37°C with 1 U of BODIPY FL phallacidin (Molecular Probes Inc, Eugene, OR). Cells were washed twice with phosphate-buffered saline, and the F-actin content was measured using a flow cytometer (FACS Calibur; Becton Dickinson, Franklin Lakes, NJ) as previously described [6] as the mean fluorescent intensity of 5,000 cells. The change in F-actin content was expressed as the percentage change from the donor value.
Adhesion molecules assay
Changes in the surface expression of L-selectin and CD11b of neutrophils were measured using flow cytometry as previously described [6]. One hundred microliters of whole blood samples were incubated for 30 minutes with 2 mg/mL of fluorescein isothiocyanate, conjugated CD62L antibody (Pharmingen, San Diego, CA), and 1 mg/mL of phycoerythrin-conjugated mouse monoclonal antihuman CD11b antibody (DAKO Laboratories, Copenhagen, Denmark) at 4°C. Identical samples were incubated with fluorescein isothiocyanate–conjugated mouse immunoglobulin G (DAKO Laboratories) and phycoerythrin-conjugated mouse immunoglobulin G2a (DAKO Laboratories) as a negative control. The erythrocytes were lysed for 60 seconds with Immuno-lyse, and leukocytes were fixed with Immuno-fix (commercial kits from Coulter Clone; Coulter Immuno, 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. The L-selectin and CD11b changes were expressed as the percentage changes compared with the donor value.
Microchannel array flow analysis
The transit time of whole blood through the microchannel array was measured as a surrogate marker of neutrophil deformability and the rheologic impact by formed microaggregates. The detailed procedures and apparatus (Microchannel Array Flow Analyzer: MC-FAN, type KH-2; Hitachi Haramachi Electronics, Co. Ltd., Hitachi, Japan) of the microchannel analysis have been described previously [7, 8]. In short, microgrooves formed in the surface of a single crystal silicon substrate were converted to leak-proof microchannels by covering them tightly with an optical flat glass plate. The contact of the two surfaces could be made watertight by mechanical pressing alone because of their optical flatness. The microgrooves in the silicon microchannel chip resembling the size of capillaries (Bloody-3S, 2600 channels, width, 6 µm, depth, 4.5 µm, length, 10 µm; Hitachi Haramachi Electronics, Co. Ltd., Hitachi, Japan) were prefilled with saline. The principal structure of the present microchannel system is shown in Figure 1.
Whole blood samples collected with 3.8% acid-citrate-dextrose were diluted with phosphate-buffered saline (1:1 by volume), and the suspension was forced to flow through the microchannels under a pressure difference of 10 cm H2O. To assess the filterability of whole blood, the transit time for each 100-µL suspension was determined. These measurements were performed immediately after blood sampling at room temperature between 20° and 25°C. Results were expressed as a percentage of the transit time of the donor samples. The blood passage through an individual channel was observed and recorded using a video microscope system.
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Fig 1. Diagram of the microchannels. The silicon substrate surface has the compartments partitioned by the banks. The contact of the upper surface of the bank to the glass plate could be made watertight with mechanical pressing alone because of the optical flatness of both surfaces. (D = depth, 4.5 µm; W = width, 6.0 µm; L = length, 10 µm; L' = length of the terrace, 30 µm.)
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Statistics
Data were analyzed using single-factor analysis of variance in a randomized block design with donor as a grouping factor. The sequential rejective Bonferroni test was used to correct for multiple comparisons [9]. All values are expressed as the mean ± standard error of the mean.
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Results
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Changes in measured blood and plasma constituents and microchannel transit times during the experiments are listed in Table 1.
Hematocrit value (data not shown) and neutrophil counts did not change significantly throughout the recirculation.
Platelet counts decreased to 25.0% ± 3.5% (p < 0.001) of the donor level after 30 minutes of recirculation and remained at the same levels until the end of recirculation. Platelet aggregation to ADP was consistently attenuated after starting recirculation and reached 34.9% ± 7.5% of the donor level at the end of 120 minutes of recirculation (p < 0.001).
Images from the video microscope (Fig 2)
and scanning electron microscope (Fig 3)
show the plugging of microchannels with neutrophils and platelet microaggregates and the disturbance of erythrocyte flow at 120 minutes of recirculation. The transit time of whole blood through microchannels increased to 185.9% ± 25.6% of the donor value at the end of 120 minutes of recirculation (Fig 4,
p = 0.0039).
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Fig 2. Video-microscope pictures before and during recirculation (donor sample and 30 minutes, 60 minutes, and 120 minutes of recirculation). The microchannels are seen in the center of each picture, and the blood flows from right to left. Plugging of microchannels with neutrophils and platelet microaggregates increased as the recirculation time increased.
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Fig 3. Scanning electron microscope picture of the microchannels. Neutrophil plugs (A) and formed platelet aggregates (B) are detectable. (Original magnification x3300 for A and x3500 for B). (N = neutrophil; P = platelet; RBC = red blood cell.)
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Fig 4. The change in transit time of whole blood through microchannels before and during recirculation. Data points are standardized as a percentage of the donor value for each time point. Values are the mean ± standard error of the mean.  p = 0.0012 at 30 minutes, p = 0.001 at 60 minutes, p = 0.0039 at 120 minutes by single-factor analysis of variance with Bonferroni correction compared with the donor value. (SC = standing control.)
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The expression of CD11b of neutrophils increased to 465.8% ± 45.2% of the donor value at 30 minutes (p < 0.001) and 576.6% ± 57.8% of the donor value at 120 minutes of recirculation (Fig 5,
p < 0.001). The expression of neutrophil L-selectin decreased consistently after recirculation was started until it reached 17.3% ± 4.9% of the donor value at 120 minutes (Fig 6,
p < 0.001). The F-actin content of neutrophil increased to 249.1% ± 60.1% of the donor value at 60 minutes of recirculation (Fig 7,
p = 0.032).
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Fig 5. Expression of CD11b on neutrophil surface before and during recirculation. Data points are standardized as a percentage of the donor value for each time point. Values are the mean ± standard error of the mean. *p < 0.001; p = 0.0087 by single-factor analysis of variance with Bonferroni correction compared with the donor value. (SC = standing control.)
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Fig 6. Expression of L-selectin on neutrophil surface before and during recirculation. Data points are standardized as a percentage of the donor value for each time point. Values are the mean ± standard error of the mean. *p < 0.001;  p = 0.014 by single-factor analysis of variance with Bonferroni correction as compared with the donor value. (SC = standing control.)
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Fig 7. Expression of neutrophil F-actin before and during recirculation. Data points are standardized as a percentage of the donor value for each time point. Values are the mean ± standard error of the mean. p = 0.014 at 30 minutes, p = 0.032 at 60 minutes by single-factor analysis of variance with Bonferroni correction as compared with the donor value. (SC = standing control.)
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Comment
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Attempts have been made to develop a method to identify actual events in the microcirculation of organs. However, there are no known algorithms for averaging spatial variations in huge heterogeneous capillary networks in the intravital methods. A single-crystal silicon substrate was first developed as a capillary model to examine blood rheology in microcirculation in 1992 [7, 8]. The silicon microchannels have three significant advantages as a capillary model. Each channel has submicrometer accuracy in dimension. A single-crystal silicon substrate can be designed to have different channel configurations. The flow behavior of blood cells can be observed microscopically in real time. The design allows the microscopically observed events in individual channels to represent the phenomenon through all the channels. Formyl-methionyl-leucyl-phenylalanine–activated neutrophils and ADP-activated platelet aggregates were observed to block the microchannels, whereas erythrocytes and erythrocyte aggregates showed easy transit through the channels [10, 11].
This study was designed to apply this newly developed microchannel technology to the rheologic evaluation of neutrophils and platelets that are stimulated by CPB. The current SECC model mimics a pediatric CPB system with a small membrane oxygenator and small-diameter tubing. A similar SECC system was used to investigate the blood–foreign surface interaction in previous studies [12, 13]. Needless to say, the SECC system lacks an endothelium, which has many molecules that function to prevent neutrophil adhesion and platelet aggregation. However, we would like to stress that this totally in vitro approach avoids the loss of activated blood cells from the circuit and also avoids the problem of new cells being recruited from the bone marrow into the circuit.
In the present study, the transit time of whole blood through microchannels was greatly prolonged with 120 minutes of SECC. Neutrophils and platelet microaggregates plugged the microchannels, as seen by video microscopic and scanning electron microscopic images. The transit of neutrophils and platelets through the microchannels, which is normally too quick to be observed, was markedly slowed on activation by SECC. Eventually the channels were blocked with neutrophils and their platelet aggregates. We suspect that loss of neutrophil deformability and formed platelet aggregates are the main factors that determine transit time. The fact that F-actin content increased while the transit time of whole blood was prolonged supports the correlation between neutrophil deformability and the transit time determined by MC-FAN, although the changes in these two factors were not exactly parallel and a huge standard deviation for F-actin at 120 minutes could make the inference uncertain. Theoretically, the changes in the neutrophil cytoskeleton with F-actin assembly at the cell periphery are thought to be responsible for the deformation of the cells [14, 15]. During neutrophil activation, rapid and significant changes occur in the amount of polymerized actin and the structure of the microfilament network inside the cells. It is not known why there is a time lag between the early decrease in platelet counts and the worsening of the transit time. During CPB, the early decrease in platelet count is generally explained by reversible adhesion on the circuit surface not by irreversible aggregation of platelets. Formation of large platelet aggregates and aggregates of platelets and other cells could be delayed for a certain period after beginning CPB.
Plasma viscosity and red blood cell deformability could be other possible factors that contribute to the transit time of whole blood through the microchannels. Despite a large dose of heparin, fibrin clots could be formed, and the deformability of red blood cells could be changed during the 120 minutes of SECC. We have not yet been able to evaluate the influence of these plasma and red blood cell factors on the transit time through the microchannels in the current SECC system, which may be a limitation in the present study. Red blood cells, however, are smaller and far more deformable than neutrophils [10, 11], although red blood cell deformability might play a certain role in such restricted flow conditions with lots of neutrophil plugs on the narrow channels. Furthermore, it is unlikely that plasma viscosity changes significantly during this stabilized simulated recirculation.
L-selectin is responsible for neutrophil rolling and margination and is shed rapidly on chemotactic stimulation. Subsequent firm adhesion to activated endothelium is mediated by the upregulation of the ß-2 integrin complex, particularly CD11b/CD18. The selectins slow neutrophils by mediating rolling; the integrins induce firm adhesion between neutrophils and endothelial cells [16]. These changes in the adhesive quality of neutrophils are important in the prolonged sequestration of neutrophils in microvessels [17]. Previous studies have found increased ß-2 integrin expression on neutrophils during simulated and clinical CPB [18–20]. The downregulation of L-selectin has been shown during simulated CPB [20] but is controversial during clinical CPB [18, 19]. In this study, SECC decreased L-selectin expression and increased the CD11b expression of neutrophils, which is compatible with previous studies using SECC and most clinical studies. The results of our adhesion molecule assay showed that SECC significantly activated neutrophils progressively during 120 minutes. We suspect that the release of L-selectin-rich neutrophils from the bone marrow may compensate for the decrease in L-selectin in clinical CPB [21]. The present silicon microchannels lack the adhesion molecules on their glass or silicon surfaces; therefore, it is unlikely that these changes of neutrophil adhesion molecules affect the microchannel transit time directly. Known adhesion mechanisms might not cause neutrophils to stick to the inorganic surfaces. The neutrophil plugging we observed on microchannels would probably occur similarly in the endothelium, and adhesion mechanisms would be added to the process of the loss of deformability. Thus, such combined process could produce tighter plugging of activated neutrophils in the real microcirculation when the inflammatory response occurs.
Cardiopulmonary bypass has profound effects on the number and function of platelets. Contact between blood and synthetic surfaces also causes a loss of platelet sensitivity to activating agents [22] and the formation of circulating platelet aggregates [23]. In vitro and in vivo studies have shown that granule membrane protein 140 on activated platelets mediated binding to neutrophils and monocytes [24] but not to lymphocytes [25]. In the present study, formed microaggregates and leukocyte-platelet conjugates as well as the neutrophil deformability changes were considered to be the main factors of impaired whole-blood filterability. It would be possible to tell that neutrophils bound with more platelets are more likely to be stuck on the narrow microchannels. Such microaggregates, conjugates, and activated neutrophils may be physically sequestered in the pulmonary or other localized vascular beds, triggering local vasoactive changes and the inflammatory reaction.
In conclusion, 120 minutes of SECC activated neutrophils and platelets. Blood contact with nonendothelial SECC surfaces caused a progressive loss in the ability of neutrophils, platelets, and their aggregates to pass through the microchannels, possibly independently of the changes in neutrophil adhesion molecules. We can test various inhibitors of neutrophils and platelets to determine whether they improve the ability of cells to pass through the simulated microcapillaries with the present SECC model and MC-FAN. MC-FAN could be an optional item for investigating the rheologic aspect of the blood–artificial surface interaction and the inflammatory response associated with CPB.
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Acknowledgments
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This work was supported by a University of Tsukuba Research Grant in 2001. The authors wish to thank Shoko Sato for her excellent technical support, Avi Landau for the language direction, and Dr Shiro Hinotsu for the statistical assistance. Some perfusion materials were provided by Mera Inc, Tokyo, Japan.
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References
|
|---|
- Edmunds L.H., Jr Inflammatory response to cardiopulmonary bypass. Ann Thorac Surg 1998;66(Suppl):S12-16.[Abstract/Free Full Text]
- Johnson D., Thomson D., Hurst T., et al. Neutrophil-mediated acute lung injury after extracorporeal perfusion. J Thorac Cardiovasc Surg 1994;107:1193-1202.[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]
- Rinder C.S., Bonan J.L., Rinder H.M., et al. Cardiopulmonary bypass induces leukocyte-platelet adhesion. Blood 1992;79:1201-1205.[Abstract/Free Full Text]
- Weerasinghe A., Taylor K.M. The platelet in cardiopulmonary bypass. Ann Thorac Surg 1998;66:2145-2152.[Abstract/Free Full Text]
- Sato Y., Hogg J.C., English D., et al. Endothelin-1 changes polymorphonuclear leukocytes’ deformability, and CD11b expression, and promotes their retention in the lung. Am J Respir Cell Mol Biol 2000;23:404-410.[Abstract/Free Full Text]
- Kikuchi Y., Sato K., Ohki H., et al. Optically accessible microchannels formed in a single-crystal silicon substrate for studies of blood rheology. Microvasc Res 1992;44:226-240.[Medline]
- Kikuchi Y., Sato K., Mizuguchi Y. Modified cell-flow microchannels in a single-crystal silicon substrate and flow behavior of blood cells. Microvasc Res 1994;47:126-139.[Medline]
- Holland B.S., Copenhaver M.D. An improved sequential rejective Bonferroni test procedure. Biometrics 1987;42:417-423.
- 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]
- Kikuchi Y. Microchannel array flow analyzer for measurement of whole blood rheology and flow characteristics of leukocytes activated by bacterial stimulation. Int Soc Opt Eng 1997;2978:165-171.
- 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-II-346.
- Addonizio V.P., Jr, Macarak E.J., Niewiarowski S., et al. Preservation of human platelets with prostaglandin Eqn 1 during in vitro simulation of cardiopulmonary bypass. Circ Res 1979;44:350-357.[Free Full Text]
- 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]
- Worthen G.S., Schwab B.D., Elson E.L., Downey G.P. Mechanics of stimulated neutrophils: cell stiffening induces retention in capillaries. Science 1989;245:183-186.[Abstract/Free Full Text]
- Carlos T.M., Harlan J.M. Leukocyte-endothelial adhesion molecules. Blood 1994;84:2068-2101.[Abstract/Free Full Text]
- Doerschuk C.M. The role of CD18-mediated adhesion in neutrophil sequestration induced by infusion of activated plasma in rabbits. Am J Respir Cell Mol Biol 1992;7:140-148.
- Le Deist F., Menasche P., Kucharski C., et al. Hypothermia during cardiopulmonary bypass delays but does not prevent neutrophil-endothelial cell adhesion. A clinical study. Circulation 1995;92:II354-358.
- Galinanes M., Watson C., Trivedi U., et al. Differential patterns of neutrophil adhesion molecules during cardiopulmonary bypass in humans. Circulation 1996;94:II364-369.
- 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]
- Van Eeden S., Miyagashima R., Haley L., Hogg J.C. L-selectin expression increases on peripheral blood polymorphonuclear leukocytes during active marrow release. Am J Respir Crit Care Med 1995;151:500-507.[Abstract/Free Full Text]
- Harker L.A., Malpass T.W., Branson H.E., et al. Mechanism of abnormal bleeding in patients undergoing cardiopulmonary bypass: acquired transient platelet dysfunction associated with selective alpha-granule release. Blood 1980;56:824-834.[Free Full Text]
- Dutton R.C., Edmunds L.H., Jr, Hutchinson J.C., et al. Platelet aggregate emboli produced in patients during cardiopulmonary bypass with membrane and bubble oxygenators and blood filters. J Thorac Cardiovasc Surg 1974;67:258-265.[Medline]
- Larsen E., Celi A., Gilbert G.E., et al. PADGEM protein: a receptor that mediates the interaction of activated platelets with neutrophils and monocytes. Cell 1989;59:305-312.[Medline]
- Moore K.L., Varki A., McEver R.P. GMP-140 binds to a glycoprotein receptor on human neutrophils: evidence for a lectin-like interaction. J Cell Biol 1991;112:491-499.[Abstract/Free Full Text]
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Abstract
Introduction
