Cell-free hemoglobin is associated with microcirculatory perfusion disturbances and acute kidney injury in rats on extracorporeal membrane oxygenation

Abstract Background Extracorporeal membrane oxygenation (ECMO) is a life-saving treatment, but carries a high risk of complications such as acute kidney injury (AKI). A contributor to AKI is hemolysis, which induces vasoconstriction and renal tubular cytotoxicity. Here, we have investigated a novel...

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Main Authors: Carolien Volleman, Dionne P. C. Dubelaar, Philippa G. Phelp, Roselique Ibelings, Anita M. Tuip-de Boer, Chantal A. Polet, Walter M. van den Bergh, Alexander P. J. Vlaar, Charissa E. van den Brom
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Language:English
Published: BMC 2025-08-01
Series:BMC Anesthesiology
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Online Access:https://doi.org/10.1186/s12871-025-03251-3
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author Carolien Volleman
Dionne P. C. Dubelaar
Philippa G. Phelp
Roselique Ibelings
Anita M. Tuip-de Boer
Chantal A. Polet
Walter M. van den Bergh
Alexander P. J. Vlaar
Charissa E. van den Brom
author_facet Carolien Volleman
Dionne P. C. Dubelaar
Philippa G. Phelp
Roselique Ibelings
Anita M. Tuip-de Boer
Chantal A. Polet
Walter M. van den Bergh
Alexander P. J. Vlaar
Charissa E. van den Brom
author_sort Carolien Volleman
collection DOAJ
description Abstract Background Extracorporeal membrane oxygenation (ECMO) is a life-saving treatment, but carries a high risk of complications such as acute kidney injury (AKI). A contributor to AKI is hemolysis, which induces vasoconstriction and renal tubular cytotoxicity. Here, we have investigated a novel hypothesis that ECMO-induced hemolysis contributes to vascular leakage, edema, microcirculatory perfusion disturbances, and AKI in a rat model. Methods Rats were exposed to 75 min of ECMO or a sham procedure as control (n = 8 per group). Hemodynamic, blood gas, and microcirculatory perfusion parameters were monitored throughout the experiment. Renal vascular leakage and edema were determined by dextran leakage (70 kDa) and wet-to-dry weight ratio. Markers of hemolysis, inflammation, endothelial activation and damage, and AKI were assessed using spectrophotometry, ELISA and Luminex. Results Initiation of ECMO increased circulating cell-free hemoglobin (CFHb) compared to baseline (4.01 vs. 1.36 OD, p < 0.001). In parallel, ECMO increased circulating levels of TNFα, IL-6, ICAM-1 and angiopoietin-2, whereas levels in the control group remained stable. The number of continuously perfused vessels (4.36 vs. 13.62 vessels/recording, p < 0.001) and the proportion of perfused vessels (PPV; 23.0 vs. 67.4%, p < 0.001) immediately decreased after initiation of ECMO when compared to controls and remained disturbed one hour after weaning from ECMO. Furthermore, NGAL, a marker of kidney injury, in plasma and urine was higher in the ECMO group compared to the controls (respectively 2191 vs. 410 ng/mL, p < 0.001; 1733 vs. 437 ng/mL, p = 0.0059). Wet-to-dry weight ratio showed increased renal edema in the group undergoing ECMO (4.50 ± 0.27 vs. 3.96 ± 0.16, p < 0.001). Moreover, increasing levels of CFHb in plasma were correlated with a decrease in PPV (r=-0.925, p < 0.001) as well as an increase in plasma NGAL (r = 0.895, p < 0.001) in rats on ECMO. Conclusion In conclusion, ECMO-induced hemolysis is paralleled by endothelial damage, microcirculatory perfusion disturbances, and kidney injury in a rat model. Our findings suggest that CFHb plays an important role in the pathophysiology of AKI, possibly via endothelial damage. Future studies should clarify the causal relationship between CFHb and endothelial damage, and explore whether targeting CFHb can improve microvascular perfusion and preserve kidney function during ECMO support.
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spelling doaj-art-eda918bc74ed4dd3aa7b702db820b1de2025-08-24T11:47:30ZengBMCBMC Anesthesiology1471-22532025-08-0125111310.1186/s12871-025-03251-3Cell-free hemoglobin is associated with microcirculatory perfusion disturbances and acute kidney injury in rats on extracorporeal membrane oxygenationCarolien Volleman0Dionne P. C. Dubelaar1Philippa G. Phelp2Roselique Ibelings3Anita M. Tuip-de Boer4Chantal A. Polet5Walter M. van den Bergh6Alexander P. J. Vlaar7Charissa E. van den Brom8Department of Intensive Care Medicine, Amsterdam UMC, University of AmsterdamDepartment of Intensive Care Medicine, Amsterdam UMC, University of AmsterdamDepartment of Intensive Care Medicine, Amsterdam UMC, University of AmsterdamDepartment of Intensive Care Medicine, Amsterdam UMC, University of AmsterdamDepartment of Intensive Care Medicine, Amsterdam UMC, University of AmsterdamDepartment of Intensive Care Medicine, Amsterdam UMC, University of AmsterdamDepartment of Critical Care, University Medical Center Groningen, University of GroningenDepartment of Intensive Care Medicine, Amsterdam UMC, University of AmsterdamDepartment of Intensive Care Medicine, Amsterdam UMC, University of AmsterdamAbstract Background Extracorporeal membrane oxygenation (ECMO) is a life-saving treatment, but carries a high risk of complications such as acute kidney injury (AKI). A contributor to AKI is hemolysis, which induces vasoconstriction and renal tubular cytotoxicity. Here, we have investigated a novel hypothesis that ECMO-induced hemolysis contributes to vascular leakage, edema, microcirculatory perfusion disturbances, and AKI in a rat model. Methods Rats were exposed to 75 min of ECMO or a sham procedure as control (n = 8 per group). Hemodynamic, blood gas, and microcirculatory perfusion parameters were monitored throughout the experiment. Renal vascular leakage and edema were determined by dextran leakage (70 kDa) and wet-to-dry weight ratio. Markers of hemolysis, inflammation, endothelial activation and damage, and AKI were assessed using spectrophotometry, ELISA and Luminex. Results Initiation of ECMO increased circulating cell-free hemoglobin (CFHb) compared to baseline (4.01 vs. 1.36 OD, p < 0.001). In parallel, ECMO increased circulating levels of TNFα, IL-6, ICAM-1 and angiopoietin-2, whereas levels in the control group remained stable. The number of continuously perfused vessels (4.36 vs. 13.62 vessels/recording, p < 0.001) and the proportion of perfused vessels (PPV; 23.0 vs. 67.4%, p < 0.001) immediately decreased after initiation of ECMO when compared to controls and remained disturbed one hour after weaning from ECMO. Furthermore, NGAL, a marker of kidney injury, in plasma and urine was higher in the ECMO group compared to the controls (respectively 2191 vs. 410 ng/mL, p < 0.001; 1733 vs. 437 ng/mL, p = 0.0059). Wet-to-dry weight ratio showed increased renal edema in the group undergoing ECMO (4.50 ± 0.27 vs. 3.96 ± 0.16, p < 0.001). Moreover, increasing levels of CFHb in plasma were correlated with a decrease in PPV (r=-0.925, p < 0.001) as well as an increase in plasma NGAL (r = 0.895, p < 0.001) in rats on ECMO. Conclusion In conclusion, ECMO-induced hemolysis is paralleled by endothelial damage, microcirculatory perfusion disturbances, and kidney injury in a rat model. Our findings suggest that CFHb plays an important role in the pathophysiology of AKI, possibly via endothelial damage. Future studies should clarify the causal relationship between CFHb and endothelial damage, and explore whether targeting CFHb can improve microvascular perfusion and preserve kidney function during ECMO support.https://doi.org/10.1186/s12871-025-03251-3ECMOExtracorporeal membrane oxygenationExtracorporeal circulationEndotheliumEdemaHemolysis
spellingShingle Carolien Volleman
Dionne P. C. Dubelaar
Philippa G. Phelp
Roselique Ibelings
Anita M. Tuip-de Boer
Chantal A. Polet
Walter M. van den Bergh
Alexander P. J. Vlaar
Charissa E. van den Brom
Cell-free hemoglobin is associated with microcirculatory perfusion disturbances and acute kidney injury in rats on extracorporeal membrane oxygenation
BMC Anesthesiology
ECMO
Extracorporeal membrane oxygenation
Extracorporeal circulation
Endothelium
Edema
Hemolysis
title Cell-free hemoglobin is associated with microcirculatory perfusion disturbances and acute kidney injury in rats on extracorporeal membrane oxygenation
title_full Cell-free hemoglobin is associated with microcirculatory perfusion disturbances and acute kidney injury in rats on extracorporeal membrane oxygenation
title_fullStr Cell-free hemoglobin is associated with microcirculatory perfusion disturbances and acute kidney injury in rats on extracorporeal membrane oxygenation
title_full_unstemmed Cell-free hemoglobin is associated with microcirculatory perfusion disturbances and acute kidney injury in rats on extracorporeal membrane oxygenation
title_short Cell-free hemoglobin is associated with microcirculatory perfusion disturbances and acute kidney injury in rats on extracorporeal membrane oxygenation
title_sort cell free hemoglobin is associated with microcirculatory perfusion disturbances and acute kidney injury in rats on extracorporeal membrane oxygenation
topic ECMO
Extracorporeal membrane oxygenation
Extracorporeal circulation
Endothelium
Edema
Hemolysis
url https://doi.org/10.1186/s12871-025-03251-3
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