Este trabajo fue apoyado por una beca de la &quot;Deutsche Forschungsgemeinschaft&quot; a SU Eisenhardt (IE 866/1-1).

<ol>
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No hay conflictos de interés declarado.

Leucocitos del endotelio interacción, la producción de especies reactivas del oxígeno y la activación del sistema del complemento son las principales características de la disfunción del tejido inducido por el IRI. 26 La microcirculación del tejido afectado es considerado como el sitio integral para el inicio inflamatoria. Aparte de los experimentos ex vivo, como los ensayos de la cámara de flujo 27, 28, es obligatorio proporcionar modelos bien establecidos de imagen intravital par...

<table border="1"><tbody><tr><td> Nombre del equipo: </td><td> Empresa: </td><td> Artículo número: </td><td> Comentarios: </td></tr><tr><td> Forene 100% (V / V) </td><td> Abad </td><td> B506 </td><td> API de isoflurano </td></tr><tr><td> Terylene sutura </td><td> Serag Weissner </td><td> OC108000 </td><td></td></tr><tr><td> Portex Bellas Diámetro tubo de polietileno </td><td> Smiths Medical </td><td> 800/100/100 </td><td> 0,28 mm de diámetro interior </td></tr><tr><td> 0,9% de solución salina </td><td> Fresinus Kabi </td><td> 808771 </td><td></td></tr><tr><td> -A cambio de punta de lujo kit de cauterio </td><td> Bovie Medical </td><td> Del1 </td><td></td></tr><tr><td> Abbocath-T 14G </td><td> Venisystems </td><td> G713- A01 </td><td> utilizado como tubo de lente </td></tr><tr><td> Servo 900C Ventilador </td><td> Maquet </td><td></td><td> utilizada como ventialtor animales </td></tr><tr><td> Transductor de presión lógico </td><td> Smiths Medical </td><td> MX1960 </td><td></td></tr><tr><td> Sirecust 404 Monitor de </td><td> Siemens </td><td></td><td></td></tr><tr><td> ABL 700 Analizador de mesa </td><td> Radiómetro </td><td></td><td> para la medición de gases en sangre </td></tr><tr><td> Almohadilla eléctrica </td><td> Effenberger </td><td> 8319 </td><td></td></tr><tr><td> Aluminio fase </td><td> ALFUN </td><td> AW7022 </td><td></td></tr><tr><td> Microscopio quirúrgico OPMI 6-SDFC </td><td> Carl Zeiss </td><td></td><td></td></tr><tr><td> Microcirugía de los instrumentos de laboratorio de set </td><td> S &amp; T </td><td> 767 </td><td></td></tr><tr><td> Biemer clip de buques </td><td> Diener </td><td> 64.562 </td><td></td></tr><tr><td> La aplicación de fórceps </td><td> Diener </td><td> 64.568 </td><td> para el clip de buques Biemer </td></tr><tr><td> Rodamina 6G </td><td> Sigma-Aldrich </td><td> R4127 </td><td></td></tr><tr><td> Vaselina blanca DAB </td><td> Winthrop </td><td> 2726853 </td><td></td></tr><tr><td> Cubreobjetos </td><td></td><td></td><td> 32x32 mm </td></tr><tr><td></td><td></td><td></td><td> Configuración intravital </td></tr><tr><td> Zeis Axio Scope A-1 MAT </td><td> Carl Zeis </td><td> 490036 </td><td> microscopio de epifluorescencia </td></tr><tr><td> 470 nm LED </td><td> Carl Zeis </td><td> 423052 </td><td> fuente de luz fluorescente </td></tr><tr><td> 2 Sistema de Colibri </td><td> Carl Zeis </td><td> 423052 </td><td></td></tr><tr><td> W Plan-Apochromat 20x / 1,0 DIC </td><td> Carl Zeis </td><td> 421452 </td><td> objetivo de inmersión en el agua </td></tr><tr><td> AxioCam MRM Rev. 3 FireWire </td><td> Carl Zeis </td><td> 426509 </td><td> cámara digital de alta resolución </td></tr><tr><td> Axio LE software de la visión </td><td> Carl Zeis </td><td> 410130 </td><td> utilizar para análisis fuera de línea </td></tr></tbody></table>

real-time digital imaging of leukocyte-endothelial interaction in ischemia-reperfusion injury (iri) of the rat cremaster muscle

La isquemia-reperfusión (IRI) ha sido implicado en una gran variedad de condiciones patológicas tales como accidente cerebrovascular, infarto de miocardio, isquemia intestinal, así como seguir. Trasplante y cirugía cardiovascular una reperfusión del tejido previamente isquémico, aunque es esencial para la prevención de la irreversible las lesiones de tejidos, provoca inflamación excesiva del tejido afectado. Adyacente a la producción de especies reactivas del oxígeno, la activación del sistema del complemento y el aumento de la permeabilidad microvascular, la activación de los leucocitos es uno de los principales actores en la cascada patológica de la lesión tisular inflamatoria durante la reperfusión. 2, 3 activación de leucocitos es un proceso de varias fases que consiste de material, la adhesión firme y la transmigración y está mediada por una compleja interacción entre las moléculas de adhesión en respuesta a factores quimiotácticos como factores del complemento, quimiocinas, o el factor activador de plaquetas. 4 &lt;p class = &quot;jove_content&quot;&gt; Mientras rodadura de leucocitos en las vénulas postcapilares está predominantemente mediado por la interacción de selectinas 5 con sus ligandos de venta libre, la adhesión firme de los leucocitos al endotelio es selectina-controlado a través de la unión a las moléculas de adhesión intercelular (ICAM) y celulares vasculares las moléculas de adhesión (VCAM). 6, 7 Estándar de oro para el estudio in vivo de leucocitos del endotelio interacción es la técnica de microscopía intravital, describió por primera vez en 1968. 8 Aunque varios modelos de IRI (isquemia-reperfusión) se han descrito para diversos órganos, 9-12 sólo unos pocos son adecuadas para la visualización directa de reclutamiento de leucocitos en el lecho microvascular en un alto nivel de calidad de imagen. 8 Estamos aquí, promover el digital de microscopía intravital epifluorescencia de la vénula postcapilares en la microcirculación cremastéricode la rata 13 como un método práctico para analizar cualitativa y cuantitativamente el reclutamiento de leucocitos por el IRI de investigación en el tejido muscular estriado y proporcionar un manual detallado para llevar a cabo la técnica. Nos ilustran aún más las trampas más comunes y proporcionar consejos útiles que deberían permitir al lector a apreciar de verdad, y con seguridad realizar el método. En un paso a paso el protocolo que describen cómo empezar con anestesia respiración controlada bajo vigilancia suficiente para evitar que el animal anestesiado con firmeza durante períodos más largos de tiempo. A continuación, describen la preparación cremastérico como una delgada lámina plana para la resolución óptica excepcional y proporcionar un protocolo para obtener imágenes de leucocitos en el IRI que ha sido bien establecido en nuestros laboratorios.

Watch this Scientific Journal Video about Real-time Digital Imaging of Leukocyte-endothelial Interaction in Ischemia-reperfusion Injury IRI of the Rat Cremaster Muscle at JoVE.com

Real-time Digital Imaging of Leukocyte-endothelial Interaction in Ischemia-reperfusion Injury IRI of the Rat Cremaster Muscle

Digital microscopía intravital epifluorescencia de vénulas post-capilares de la microcirculación cremastérico es un método conveniente para obtener información sobre la interacción leucocito-endotelio<em&gt; En vivo</em&gt; En la lesión por isquemia-reperfusión (IRI) de músculo estriado. Estamos aquí proporcionar un protocolo detallado para realizar con seguridad la técnica y discutir sus aplicaciones y limitaciones.

En tiempo real de imágenes digitales de los leucocitos del endotelio interacción en el daño por isquemia-reperfusión (IRI) del músculo cremáster Rata

Ischemia-reperfusion injury (IRI) has been implicated in a large array of pathological conditions such as cerebral stroke, myocardial infarction, ...

real-time-digital-imaging-leukocyte-endothelial-interaction-ischemia

Research

JoVE Journal

Medicine

Real-time Digital Imaging of Leukocyte-endothelial Interaction in Ischemia-reperfusion Injury (IRI) of the Rat Cremaster Muscle

15.0K vistas.  University of Freiburg Medical Centre. Digital microscopía intravital epifluorescencia de vénulas post-capilares de la microcirculación cremastérico es un método conveniente para obtener información sobre la interacción leucocito-endotelio En vivo En la lesión por isquemia-reperfusión (IRI) de músculo estriado. Estamos aquí proporcionar un protocolo detallado para realizar con seguridad la técnica y discutir sus aplicaciones y limitaciones.

Video: Real-time Digital Imaging of Leukocyte-endothelial Interaction in Ischemia-reperfusion Injury IRI of the Rat Cremaster Muscle

In situ Transverse Rectus Abdominis Myocutaneous Flap: A Rat Model of Myocutaneous Ischemia Reperfusion Injury

Free tissue transfer is the gold standard of reconstructive surgery to repair complex defects not amenable to local options or those requiring composite tissue. Ischemia reperfusion injury (IRI) is a known cause of partial free flap failure and has no effective treatment. Establishing a laboratory model of this injury can prove costly both financially as larger mammals are conventionally used and in the expertise required by the technical difficulty of these procedures typically requires employing an experienced microsurgeon. This publication and video demonstrate the effective use of a model of IRI in rats which does not require microsurgical expertise. This procedure is an in situ model of a transverse abdominis myocutaneous (TRAM) flap where atraumatic clamps are utilized to reproduce the ischemia-reperfusion injury associated with this surgery. A laser Doppler Imaging (LDI) scanner is employed to assess flap perfusion and the image processing software, Image J to assess percentage area skin survival as a primary outcome measure of injury.

In situ Transverse Rectus Abdominis Myocutaneous Flap: A Rat Model of Myocutaneous Ischemia Reperfusion Injury

Free tissue transfer is widely employed in reconstructive surgery to restore form and function following oncological resection and trauma. Preconditioning this tissue prior to surgery may improve outcome. This article describes an in situ transverse rectus abdominis myocutaneous flap (TRAM) in rats as a means for testing preconditioning strategies.

In situ recto abdominal transverso colgajo miocutáneo: un modelo de rata de miocutáneo isquemia reperfusión

Free tissue transfer is the gold standard of reconstructive surgery to repair complex defects not amenable to local options or those requiring composite ...

Investigación

Medicina

Ischemia-reperfusion Model of Acute Kidney Injury and Post Injury Fibrosis in Mice

Ischemia-reperfusion induced acute kidney injury (IR-AKI) is widely used as a model of AKI in mice, but results are often quite variable with high, often unreported mortality rates that may confound analyses. Bilateral renal pedicle clamping is commonly used to induce IR-AKI, but differences between effective clamp pressures and/or renal responses to ischemia between kidneys often lead to more variable results. In addition, shorter clamp times are known to induce more variable tubular injury, and while mice undergoing bilateral injury with longer clamp times develop more consistent tubular injury, they often die within the first 3 days after injury due to severe renal insufficiency. To improve post-injury survival and obtain more consistent and predictable results, we have developed two models of unilateral ischemia-reperfusion injury followed by contralateral nephrectomy. Both surgeries are performed using a dorsal approach, reducing surgical stress resulting from ventral laparotomy, commonly used for mouse IR-AKI surgeries. For induction of moderate injury BALB/c mice undergo unilateral clamping of the renal pedicle for 26 min and also undergo simultaneous contralateral nephrectomy. Using this approach, 50-60% of mice develop moderate AKI 24 hr after injury but 90-100% of mice survive. To induce more severe AKI, BALB/c mice undergo renal pedicle clamping for 30 min followed by contralateral nephrectomy 8 days after injury. This allows functional assessment of renal recovery after injury with 90-100% survival. Early post-injury tubular damage as well as post injury fibrosis are highly consistent using this model.

We describe models of moderate and severe ischemia-reperfusion-induced kidney injury in which the mice undergo unilateral renal pedicle clamping followed by simultaneous or delayed contralateral nephrectomy, respectively. These models consistently give rise to renal dysfunction and post-injury fibrosis, but injury severity and survival are dependent on mouse background, age and surgical equipment.

La isquemia-reperfusión modelo de lesión renal aguda y post lesión fibrosis en ratones

Ischemia-reperfusion induced acute kidney injury (IR-AKI) is widely used as a model of AKI in mice, but results are often quite variable with high, often ...

Induction and Assessment of Ischemia-reperfusion Injury in Langendorff-perfused Rat Hearts

The biochemical events surrounding ischemia reperfusion injury in the acute setting are of great importance to furthering novel treatment options for myocardial infarction and cardiac complications of thoracic surgery. The ability of certain drugs to precondition the myocardium against ischemia reperfusion injury has led to multiple clinical trials, with little success. The isolated heart model allows acute observation of the functional effects of ischemia reperfusion injury in real time, including the effects of various pharmacological interventions administered at any time-point before or within the ischemia-reperfusion injury window. Since brief periods of ischemia can precondition the heart against ischemic injury, in situ aortic cannulation is performed to allow for functional assessment of non-preconditioned myocardium. A saline filled balloon is placed into the left ventricle to allow for real-time measurement of pressure generation. Ischemic injury is simulated by the cessation of perfusion buffer flow, followed by reperfusion. The duration of both ischemia and reperfusion can be modulated to examine biochemical events at any given time-point. Although the Langendorff isolated heart model does not allow for the consideration of systemic events affecting ischemia and reperfusion, it is an excellent model for the examination of acute functional and biochemical events within the window of ischemia reperfusion injury as well as the effect of pharmacological intervention on cardiac pre- and postconditioning. The goal of this protocol is to demonstrate how to perform in situ aortic cannulation and heart excision followed by ischemia/reperfusion injury in the Langendorff model.

The isolated rat heart is an enduring model for ischemia reperfusion injury. Here, we describe the process of harvesting the beating heart from a rat via in situ aortic cannulation, Langendorff perfusion of the heart, simulated ischemia-reperfusion injury, and infarct staining to confirm the extent of ischemic insult.

Inducción y Evaluación de la isquemia-reperfusión en perfundidos Langendorff corazones de ratas

The biochemical events surrounding ischemia reperfusion injury in the acute setting are of great importance to furthering novel treatment options for ...

Murine Model of Intestinal Ischemia-reperfusion Injury

Intestinal ischemia is a life-threatening condition associated with a broad range of clinical conditions including atherosclerosis, thrombosis, hypotension, necrotizing enterocolitis, bowel transplantation, trauma and chronic inflammation. Intestinal ischemia-reperfusion (IR) injury is a consequence of acute mesenteric ischemia, caused by inadequate blood flow through the mesenteric vessels, resulting in intestinal damage. Reperfusion following ischemia can further exacerbate damage of the intestine. The mechanisms of IR injury are complex and poorly understood. Therefore, experimental small animal models are critical for understanding the pathophysiology of IR injury and the development of novel therapies.
Here we describe a mouse model of acute intestinal IR injury that provides reproducible injury of the small intestine without mortality. This is achieved by inducing ischemia in the region of the distal ileum by temporally occluding the peripheral and terminal collateral branches of the superior mesenteric artery for 60 min using microvascular clips. Reperfusion for 1 hr, or 2 hr after injury results in reproducible injury of the intestine examined by histological analysis. Proper position of the microvascular clips is critical for the procedure. Therefore the video clip provides a detailed visual step-by-step description of this technique. This model of intestinal IR injury can be utilized to study the cellular and molecular mechanisms of injury and regeneration.

Here we describe the detailed procedure of intestinal ischemia-reperfusion in mice which results in reproducible injury without mortality to encourage the standardization of this technique across the field. This model of intestinal ischemia-reperfusion injury can be utilized to study the cellular and molecular mechanisms of injury and regeneration.

Modelo murino de isquemia-reperfusión intestinal Lesiones

Intestinal ischemia is a life-threatening condition associated with a broad range of clinical conditions including atherosclerosis, thrombosis, ...

Inducing Ischemia-reperfusion Injury in the Mouse Ear Skin for Intravital Multiphoton Imaging of Immune Responses

Ischemia-reperfusion injury (IRI) occurs when there is transient hypoxia due to the obstruction of blood flow (ischemia) followed by a subsequent re-oxygenation of the tissues (reperfusion). In the skin, ischemia-reperfusion (IR) is the main contributing factor to the pathophysiology of pressure ulcers. While the cascade of events leading up to the inflammatory response has been well studied, the spatial and temporal responses of the different subsets of immune cells to an IR injury are not well understood. Existing models of IR using the clamping technique on the skin flank are highly invasive and unsuitable for studying immune responses to injury, while similar non-invasive magnet clamping studies in the skin flank are less-than-ideal for intravital imaging studies. In this protocol, we describe a robust model of non-invasive IR developed on mouse ear skin, where we aim to visualize in real-time the cellular response of immune cells after reperfusion via multiphoton intravital imaging (MP-IVM).

This protocol describes the induction of an ischemia-reperfusion (IR) model on mouse ear skin using magnet clamping. Using a custom-built intravital imaging model, we study in vivo inflammatory responses post-reperfusion. The rationale behind the development of this technique is to extend the understanding of how leukocytes respond to skin IR injury.

La inducción de la lesión por isquemia-reperfusión en la piel de la oreja del ratón para intravital n fotones de imágenes de la respuesta inmune

Ischemia-reperfusion injury (IRI) occurs when there is transient hypoxia due to the obstruction of blood flow (ischemia) followed by a subsequent ...

Evaluation of Coronary Flow Reserve After Myocardial Ischemia Reperfusion in Rats

Coronary artery disease is the leading cause of death worldwide. After an acute myocardial infarction, early and successful myocardial intervention via recanalization of the coronary artery is the most effective strategy for reducing the size of ischemic myocardium. The coronary microvasculature cannot be visualized and imaged&#160;in vivo, but there are several invasive and noninvasive techniques that can be used to assess parameters which depend directly on coronary microvascular function. The endothelial function after ischemia reperfusion can be assessed also at the level of the coronary circulation via the coronary flow reserve (CFR).&#160;In this study, peak velocity of left anterior descending (LAD) coronary arteries was measured in rats in vivo via Transthoracic Doppler Echocardiography during resting and stress challenge (induced by Dobutamine). A normal heart can increase its coronary blood flow up to four times above the resting values during stress induction. Following ischemia reperfusion, we found a significantly diminished CFR, which can be used as a marker of coronary microvascular dysfunction. CFR has opened a window on the importance of microvascular dysfunction and has been shown to predict cardiovascular risk independent of whether the severe obstructive disease is present.

Coronary flow reserve (CFR), is defined as the ratio of maximal coronary blood flow to the resting coronary blood flow. We present a protocol for evaluating CFR in rats via ultrasound, which offers the opportunity to predict cardiovascular risk factors in the absence of obstructive coronary disease.

Evaluación de la Reserva de Flujo Coronario Después de la Reperfusión de Isquemia Miocárdica en Ratas

Coronary artery disease is the leading cause of death worldwide. After an acute myocardial infarction, early and successful myocardial intervention via ...

A Pre-clinical Rat Model for the Study of Ischemia-reperfusion Injury in Reconstructive Microsurgery

Ischemia-reperfusion injury is the main cause of flap failure in reconstructive microsurgery. The rat is the preferred preclinical animal model in many areas of biomedical research due to its cost-effectiveness and its translation to humans. This protocol describes a method to create a preclinical free skin flap model in rats with ischemia-reperfusion injury. The described 3 cm x 6 cm rat free skin flap model is easily obtained after the placement of several vascular ligatures and the section of the vascular pedicle. Then, 8 h after the ischemic insult and completion of the microsurgical anastomosis, the free skin flap develops the tissue damage. These ischemia-reperfusion injury-related damages can be studied in this model, making it a suitable model for evaluating therapeutic agents to address this pathophysiological process. Furthermore, two main monitoring techniques are described in the protocol for the assessment of this animal model: transit-time ultrasound technology and laser speckle contrast analysis.

Here, we describe a pre-clinical animal model for studying the pathophysiology of ischemia-reperfusion injury in reconstructive microsurgery. This free skin flap model based on the superficial caudal epigastric vessels in the rat may also allow for the evaluation of different therapies and compounds to counteract ischemia-reperfusion injury-related damage.

Un modelo preclínico de rata para el estudio de la lesión por isquemia-reperfusión en microcirugía reconstructiva

Ischemia-reperfusion injury is the main cause of flap failure in reconstructive microsurgery. The rat is the preferred preclinical animal model in many ...

Delayed Intramyocardial Delivery of Stem Cells after Ischemia Reperfusion Injury in a Murine Model

There is significant interest in the use of stem cells (SCs) for the recovery of cardiac function in individuals with myocardial injuries. Most commonly, cardiac stem cell therapy is studied by delivering SCs concurrently with the induction of myocardial injury. However, this approach presents two significant limitations: the early hostile pro-inflammatory ischemic environment may affect the survival of transplanted SCs, and it does not represent the subacute infarction scenario where SCs will likely be used. Here we describe a two-part series of surgical procedures for the induction of ischemia-reperfusion injury and delivery of mesenchymal stem cells (MSCs). This method of stem cell administration may allow for the longer viability and retention around damaged tissue by circumventing the initial immune response. A model of ischemia reperfusion injury was induced in mice accompanied by the delivery of mesenchymal stem cells (3.0 x 105), stably expressing the reporter gene firefly luciferase under the constitutively expressed CMV promoter, intramyocardially 7 days later. The animals were imaged via ultrasound and bioluminescent imaging for confirmation of injury and injection of cells, respectively. Importantly, there was no added complication rate when performing this two-procedure approach for SC delivery. This method of stem cell administration, collectively with the utilization of state-of-the-art reporter genes, may allow for the in vivo study of viability and retention of transplanted SCs in a situation of chronic ischemia commonly seen clinically, while also circumventing the initial pro-inflammatory response. In summary, we established a protocol for the delayed delivery of stem cells into the myocardium, which can be used as a potential new approach in promoting regeneration of the damaged tissue.

Stems cells are continuously investigated as potential treatments for individuals with myocardial damage, however, their decreased viability and retention within injured tissue can impact their long-term efficacy. In this manuscript we describe an alternative method for stem cell delivery in a murine model of ischemia reperfusion injury.

Retraso en la administración intramiocardio de células madre después de la lesión por reperfusión de isquemia en un modelo murino

There is significant interest in the use of stem cells (SCs) for the recovery of cardiac function in individuals with myocardial injuries. Most commonly, ...

Organ Ischemia-Reperfusion Injury by Simulating Hemodynamic Changes in Rat Liver Transplant Model

Orthotopic liver transplantation (OLT) in rats is a tried and proven animal model used for preoperative, intraoperative, and postoperative studies, including ischemia-reperfusion injury (IRI) of extrahepatic organs. This model requires numerous experiments and devices. The duration of anhepatic phase is closely related to the time to develop IRI after transplantation. In this experiment, we used hemodynamic changes to induce extrahepatic organ damage in rats and determined the maximum tolerance time. The time until the most severe organ injury varied for different organs. This method can easily be replicated and can also be used to study IRI of the extrahepatic organs after liver transplantation.

This paper provides a detailed description of how to build an animal model of the anhepatic phase (liver ischemia) in rats to facilitate basic research into ischemia-reperfusion injury after liver transplantation.

Lesión de isquemia-reperfusión de órganos mediante la simulación de cambios hemodinámicos en el modelo de trasplante de hígado de rata

Orthotopic liver transplantation (OLT) in rats is a tried and proven animal model used for preoperative, intraoperative, and postoperative studies, ...

Real-Time Assessment of Spinal Cord Microperfusion in a Porcine 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 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.

Evaluación en tiempo real de la microperfusión de la médula espinal en un modelo porcino de isquemia/reperfusión

Spinal cord injury is a devastating complication of aortic repair. Despite developments for the prevention and treatment of spinal cord injury, its ...