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Representative Results






Real-Time Assessment of Spinal Cord Microperfusion in a Porcine Model of Ischemia/Reperfusion

Published: December 10th, 2020



1Department of Anesthesiology, Center of Anesthesiology and Intensive Care Medicine, University Medical Center Hamburg-Eppendorf, 2University Department for Vascular Surgery and Department of Operative Medicine, Medical University of Innsbruck, 3Department of Medical Biometry and Epidemiology, University Medical Center Hamburg-Eppendorf, 4Department of Vascular Medicine, University Heart and Vascular Center Hamburg (UHZ), 5Department of Cardiology, Rostock University Medical Center, 6University Department for Cardiac Surgery, Heart Center Leipzig

Spinal cord microcirculation plays a pivotal role in spinal cord injury. Most methods do not allow real-time assessment of spinal cord microcirculation, which is essential for the development of microcirculation-targeted therapies. Here, we propose a protocol using Laser-Doppler-Flow Needle probes in a large animal model of ischemia/reperfusion.

Spinal cord injury is a devastating complication of aortic repair. Despite developments for the prevention and treatment of spinal cord injury, its incidence is still considerably high and therefore, influences patient outcome. Microcirculation plays a key role in tissue perfusion and oxygen supply and is often dissociated from macrohemodynamics. Thus, direct evaluation of spinal cord microcirculation is essential for the development of microcirculation-targeted therapies and the evaluation of existing approaches in regard to spinal cord microcirculation. However, most of the methods do not provide real-time assessment of spinal cord microcirculation. The aim of this study is to describe a standardized protocol for real-time spinal cord microcirculatory evaluation using laser-Doppler needle probes directly inserted in the spinal cord. We used a porcine model of ischemia/reperfusion to induce deterioration of the spinal cord microcirculation. In addition, a fluorescent microsphere injection technique was used. Initially, animals were anesthetized and mechanically ventilated. Thereafter, laser-Doppler needle probe insertion was performed, followed by the placement of cerebrospinal fluid drainage. A median sternotomy was performed for exposure of the descending aorta to perform aortic cross-clamping. Ischemia/reperfusion was induced by supra-celiac aortic cross-clamping for a total of 48 min, followed by reperfusion and hemodynamic stabilization. Laser-Doppler Flux was performed in parallel with macrohemodynamic evaluation. In addition, automated cerebrospinal fluid drainage was used to maintain a stable cerebrospinal pressure. After completion of the protocol, animals were sacrificed, and the spinal cord was harvested for histopathological and microsphere analysis. The protocol reveals the feasibility of spinal cord microperfusion measurements using laser-Doppler probes and shows a marked decrease during ischemia as well as recovery after reperfusion. Results showed comparable behavior to fluorescent microsphere evaluation. In conclusion, this new protocol might provide a useful large animal model for future studies using real-time spinal cord microperfusion assessment in ischemia/reperfusion conditions.

Spinal cord injury induced by ischemia/reperfusion (SCI) is one of the most devastating complications of aortic repair associated with reduced outcome1,2,3,4. Current prevention and treatment options for SCI include the optimization of macrohemodynamic parameters as well as the normalization of cerebrospinal fluid pressure (CSP) to improve spinal cord perfusion pressure2,5,6,7,8

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The study was approved by the Governmental Commission on the Care and Use of Animals of the City of Hamburg (Reference-No. 60/17). The animals received care in compliance with the 'Guide for the Care and Use of Laboratory Animals' (NIH publication No. 86-23, revised 2011) as well as FELASA recommendations and experiments were carried out according to the ARRIVE guidelines24,25. This study was an acute trial, and all animals were euthanized at the end of p.......

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All six animals survived until the completion of the protocol. Animal weight was 48.2 ± 2.9 kg; five animals were male, and one animal was female. Spinal cord needle probe insertion as well as spinal cord Flux measurement was feasible in all animals.

Examples of real-time spinal cord microcirculatory recordings in combination with cerebral microcirculatory and macrohemodynamic recordings during aortic cross-clamping for isc.......

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SCI induced by spinal cord ischemia is a major complication of aortic repair with tremendous impact on patient outcome1,2,3,4,10,11,12. Microcirculation-targeted therapies to prevent and treat SCI are most promising. The protocol provides a reproducible method for real-time spinal cord micro.......

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The authors would like to thank Lena Brix, V.M.D, Institute of Animal Research, Hannover Medical School, as well as Mrs. Jutta Dammann, Facility of Research Animal Care, University Medical Center Hamburg-Eppendorf, Germany, for providing pre- and perioperative animal care and their technical assistance on animal handling. The authors would further like to thank Dr. Daniel Manzoni, Department of Vascular Surgery, Hôpital Kirchberg, Luxembourg, for his technical assistance.


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Name Company Catalog Number Comments
CardioMed Flowmeter Medistim AS, Oslo, Norway CM4000 Flowmeter for Flow-Probe Femoral Artery
CardioMed Flow-Probe, 5mm Medistim AS, Oslo, Norway PS100051 Flow-Probe Femoral Artery
COnfidence probe,  Transonic Systems Inc., Ithaca, NY, USA MA16PAU Flow-Probe Aorta
16 mm liners
DIVA Sevoflurane Vapor Dräger Medical, Lübeck, Germany Vapor
Hotline Level 1 Fluid Warmer Smiths Medical Germany GmbH, Grasbrunn, Germany HL-90-DE-230 Fluid Warmer
Infinity Delta Dräger Medical, Lübeck, Germany Basic Monitoring Hardware
Infinity Hemo Dräger Medical, Lübeck, Germany Basic Pressure Monitoring and Pulmonary Thermodilution Hardware
LabChart Pro ADInstruments Ltd., Oxford, UK v8.1.16 Synchronic Laser-Doppler, Blood Pressure, ECG and Blood-Flow Aquisition Software
LiquoGuard 7 Möller Medical GmbH, Fulda, Germany Cerebrospinal Fluid Drainage System
Millar Micro-Tip Pressure Catheter (5F, Single, Curved, 120cm, PU/WD) ADInstruments Ltd., Oxford, UK SPR-350 Pressure-Tip Catheter Aorta
moor VMS LDF moor Instruments, Devon, UK Designated Laser-Doppler Hardware
moor VMS Research Software moor Instruments, Devon, UK Designated Laser-Doppler Software
Perivascular Flow Module Transonic Systems Inc., Ithaca, NY, USA TS 420 Flow-Module for Flow-Probe Aorta
PiCCO 2, Science Version Getinge AB, Göteborg, Sweden v. 6.0 Blood Pressure and Transcardiopulmonary Monitoring Hard- and Software
PiCCO 5 Fr. 20cm Getinge AB, Göteborg, Sweden Thermistor-tipped Arterial Line 
PowerLab ADInstruments Ltd., Oxford, UK PL 3516 Synchronic Laser-Doppler, Blood Pressure, ECG and Blood-Flow Aquisition Hardware
QuadBridgeAmp ADInstruments Ltd., Oxford, UK FE 224 Four Channel Bridge Amplifier for Laser-Doppler and Invasive Blood Pressure Aquisition
Silverline Spiegelberg, Hamburg, Germany ELD33.010.02 Cerebrospinal Fluid Drainage
SPSS statistical software package  IBM SPSS Statistics Inc., Armonk, New York, USA v. 27 Statistical Software
Twinwarm Warming System Moeck & Moeck GmbH, Hamburg, Germany 12TW921DE Warming System
Universal II Warming Blanket Moeck & Moeck GmbH, Hamburg, Germany 906 Warming Blanket
VP 3 Probe, 8mm length (individually manufactured) moor Instruments, Devon, UK Laser-Doppler Probe
Zeus Dräger Medical, Lübeck, Germany Anesthesia Machine

  1. Etz, C. D., et al. Contemporary spinal cord protection during thoracic and thoracoabdominal aortic surgery and endovascular aortic repair: a position paper of the vascular domain of the European Association for Cardio-Thoracic Surgerydagger. The European Journal of Cardio-Thoracic Surgery. 47 (6), 943-957 (2015).
  2. Schraag, S. Postoperative management. Best Practice & Research Clinical Anaesthesiology. 30 (3), 381-393 (2016).
  3. Cambria, R. P., et al. Thoracoabdominal aneurysm repair: results with 337 operations performed over a 15-year interval. Annals of Surgery. 236 (4), 471-479 (2002).
  4. Becker, D. A., McGarvey, M. L., Rojvirat, C., Bavaria, J. E., Messe, S. R. Predictors of outcome in patients with spinal cord ischemia after open aortic repair. Neurocritical Care. 18 (1), 70-74 (2013).
  5. McGarvey, M. L., et al. The treatment of spinal cord ischemia following thoracic endovascular aortic repair. Neurocritical Care. 6 (1), 35-39 (2007).
  6. Fukui, S., et al. Development of collaterals to the spinal cord after endovascular stent graft repair of thoracic aneurysms. European Journal of Vascular and Endovascular Surgery. 52 (6), 801-807 (2016).
  7. Augoustides, J. G., Stone, M. E., Drenger, B. Novel approaches to spinal cord protection during thoracoabdominal aortic interventions. Current Opinion in Anesthesiology. 27 (1), 98-105 (2014).
  8. Bicknell, C. D., Riga, C. V., Wolfe, J. H. Prevention of paraplegia during thoracoabdominal aortic aneurysm repair. European Journal of Vascular and Endovascular Surgery. 37 (6), 654-660 (2009).
  9. Feezor, R. J., Lee, W. A. Strategies for detection and prevention of spinal cord ischemia during TEVAR. Seminars in Vascular Surgery. 22 (3), 187-192 (2009).
  10. Heidemann, F., et al. Incidence, predictors, and outcomes of spinal cord ischemia in elective complex endovascular aortic repair: An analysis of health insurance claims. Journal of Vascular Surgery. , (2020).
  11. Rizvi, A. Z., Sullivan, T. M. Incidence, prevention, and management in spinal cord protection during TEVAR. Journal of Vascular Surgery. 52 (4), 86-90 (2010).
  12. Wortmann, M., Bockler, D., Geisbusch, P. Perioperative cerebrospinal fluid drainage for the prevention of spinal ischemia after endovascular aortic repair. Gefasschirurgie. 22, 35-40 (2017).
  13. Saugel, B., Trepte, C. J., Heckel, K., Wagner, J. Y., Reuter, D. A. Hemodynamic management of septic shock: is it time for "individualized goal-directed hemodynamic therapy" and for specifically targeting the microcirculation. Shock. 43 (6), 522-529 (2015).
  14. Moore, J. P., Dyson, A., Singer, M., Fraser, J. Microcirculatory dysfunction and resuscitation: why, when, and how. British Journal of Anaesthesia. 115 (3), 366-375 (2015).
  15. De Backer, D., Creteur, J., Preiser, J. C., Dubois, M. J., Vincent, J. L. Microvascular blood flow is altered in patients with sepsis. American Journal of Respiratory and Critical Care Medicine. 166 (1), 98-104 (2002).
  16. De Backer, D., Creteur, J., Dubois, M. J., Sakr, Y., Vincent, J. L. Microvascular alterations in patients with acute severe heart failure and cardiogenic shock. American Heart Journal. 147 (1), 91-99 (2004).
  17. Sakr, Y., Dubois, M. J., De Backer, D., Creteur, J., Vincent, J. L. Persistent microcirculatory alterations are associated with organ failure and death in patients with septic shock. Critical Care Medicine. 32 (9), 1825-1831 (2004).
  18. Trzeciak, S., et al. Early microcirculatory perfusion derangements in patients with severe sepsis and septic shock: relationship to hemodynamics, oxygen transport, and survival. Annals of Emergency Medicine. 49 (1), 88-98 (2007).
  19. Donati, A., et al. From macrohemodynamic to the microcirculation. Critical Care Research and Practice. 2013, 892710 (2013).
  20. Hamamoto, Y., Ogata, T., Morino, T., Hino, M., Yamamoto, H. Real-time direct measurement of spinal cord blood flow at the site of compression: relationship between blood flow recovery and motor deficiency in spinal cord injury. Spine. 32 (18), 1955-1962 (2007).
  21. Soubeyrand, M., et al. Real-time and spatial quantification using contrast-enhanced ultrasonography of spinal cord perfusion during experimental spinal cord injury. Spine. 37 (22), 1376-1382 (2012).
  22. Han, S., et al. Rescuing vasculature with intravenous angiopoietin-1 and alpha v beta 3 integrin peptide is protective after spinal cord injury. Brain. 133, 1026-1042 (2010).
  23. Muradov, J. M., Ewan, E. E., Hagg, T. Dorsal column sensory axons degenerate due to impaired microvascular perfusion after spinal cord injury in rats. Experimental Neurology. 249, 59-73 (2013).
  24. Guillen, J., , . FELASA guidelines and recommendations. J Am Assoc Lab Anim Sci. 51, 311-321 (2012).
  25. Kilkenny, C., Browne, W. J., Cuthill, I. C., Emerson, M., Altman, D. G. Improving bioscience research reporting: the ARRIVE guidelines for reporting animal research. Osteoarthritis Cartilage. 20, 256-260 (2012).
  26. Ospina-Tascon, G., et al. Effects of fluids on microvascular perfusion in patients with severe sepsis. Intensive Care Medicine. 36 (6), 949-955 (2010).
  27. Pottecher, J., et al. Both passive leg raising and intravascular volume expansion improve sublingual microcirculatory perfusion in severe sepsis and septic shock patients. Intensive Care Medicine. 36 (11), 1867-1874 (2010).
  28. De Backer, D., Ortiz, J. A., Salgado, D. Coupling microcirculation to systemic hemodynamics. Current Opinion in Critical Care. 16 (3), 250-254 (2010).
  29. van Genderen, M. E., et al. Microvascular perfusion as a target for fluid resuscitation in experimental circulatory shock. Critical care medicine. 42 (2), 96-105 (2014).
  30. Ince, C. Hemodynamic coherence and the rationale for monitoring the microcirculation. Critical care. 19, 8 (2015).
  31. Kise, Y., et al. Directly measuring spinal cord blood flow and spinal cord perfusion pressure via the collateral network: correlations with changes in systemic blood pressure. Journal of Thoracic and Cardiovascular Surgery. 149 (1), 360-366 (2015).
  32. Haunschild, J., et al. Detrimental effects of cerebrospinal fluid pressure elevation on spinal cord perfusion: first-time direct detection in a large animal model. European Journal of Cardio-Thoracic Surgery. 58 (2), 286-293 (2020).
  33. Wipper, S., et al. Impact of hybrid thoracoabdominal aortic repair on visceral and spinal cord perfusion: The new and improved SPIDER-graft. Journal of Thoracic and Cardiovascular Surgery. 158 (3), 692-701 (2019).
  34. Kluttig, R., et al. Invasive hemodynamic monitoring of aortic and pulmonary artery hemodynamics in a large animal model of ARDS. Journal of Visualized Experiments. (141), e57405 (2018).
  35. Detter, C., et al. Fluorescent cardiac imaging: a novel intraoperative method for quantitative assessment of myocardial perfusion during graded coronary artery stenosis. Circulation. 116 (9), 1007-1014 (2007).
  36. Wipper, S., et al. Distinction of non-ischemia inducing versus ischemia inducing coronary stenosis by fluorescent cardiac imaging. International Journal of Cardiovascular Imaging. 32 (2), 363-371 (2016).
  37. Etz, C. D., et al. Spinal cord blood flow and ischemic injury after experimental sacrifice of thoracic and abdominal segmental arteries. European Journal of Cardio-Thoracic Surgery. 33 (6), 1030-1038 (2008).
  38. Saugel, B., Scheeren, T. W. L., Teboul, J. L. Ultrasound-guided central venous catheter placement: a structured review and recommendations for clinical practice. Critical care. 21 (1), 225 (2017).
  39. Marty, B., et al. Partial inflow occlusion facilitates accurate deployment of thoracic aortic endografts. Journal of Endovascular Therapy. 11 (2), 175-179 (2004).
  40. Matyal, R., et al. Monitoring the variation in myocardial function with the Doppler-derived myocardial performance index during aortic cross-clamping. Journal of Cardiothoracic and Vascular Anesthesia. 26 (2), 204-208 (2012).
  41. Miller, R. D. . Miller'sanesthesia. 8th Edition. , (2015).
  42. Martikos, G., et al. Remote ischemic preconditioning decreases the magnitude of hepatic ischemia-reperfusion injury on a swine model of supraceliac aortic cross-clamping. Annals of Vascular Surgery. 48, 241-250 (2018).
  43. Lazaris, A. M., et al. Protective effect of remote ischemic preconditioning in renal ischemia/reperfusion injury, in a model of thoracoabdominal aorta approach. Journal of Surgical Research. 154 (2), 267-273 (2009).
  44. Ince, C., et al. Second consensus on the assessment of sublingual microcirculation in critically ill patients: results from a task force of the European Society of Intensive Care Medicine. Intensive Care Medicine. 44 (3), 281-299 (2018).
  45. Edul, V. S., et al. Dissociation between sublingual and gut microcirculation in the response to a fluid challenge in postoperative patients with abdominal sepsis. Annals of intensive care. 4, 39 (2014).
  46. Schierling, W., et al. Sonographic real-time imaging of tissue perfusion in a porcine haemorrhagic shock model. Ultrasound in Medicine and Biology. 45 (10), 2797-2804 (2019).
  47. Jing, Y., Bai, F., Chen, H., Dong, H. Using Laser Doppler Imaging and Monitoring to Analyze Spinal Cord Microcirculation in Rat. Journal of Visualized Experiments. (135), e56243 (2018).
  48. Jing, Y., Bai, F., Chen, H., Dong, H. Meliorating microcirculatory with melatonin in rat model of spinal cord injury using laser Doppler flowmetry. Neuroreport. 27 (17), 1248-1255 (2016).
  49. Jing, Y., Bai, F., Chen, H., Dong, H. Melatonin prevents blood vessel loss and neurological impairment induced by spinal cord injury in rats. Journal of Spinal Cord Medicine. 40 (2), 222-229 (2017).
  50. Phillips, J. P., Cibert-Goton, V., Langford, R. M., Shortland, P. J. Perfusion assessment in rat spinal cord tissue using photoplethysmography and laser Doppler flux measurements. Journal of Biomedical Optics. 18 (3), 037005 (2013).
  51. Glenny, R. W., Bernard, S. L., Lamm, W. J. Hemodynamic effects of 15-microm-diameter microspheres on the rat pulmonary circulation. Journal of Applied Physiology. 89 (1985), 499-504 (2000).

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