1. Procedimientos de ratón anestesia <ol><li> Los animales son extraídas de su jaula y se colocan en una cámara de inducción de la anestesia antes de la operación para la inducción de gas en isoflurano 5% y el 100% de oxígeno, con un caudal de 0,5 litros por minuto (vaporizador controlada). </li><li> Los ratones son sedados en la cámara de inducción de la anestesia, se retiran de la cámara y la parte ventral del abdomen se afeita con una maquinilla eléctrica, o mantenerse dentro de la cámara si es una crema depilatoria el cabello se utiliza (1 minuto). </li><li> Si bien todavía bajo sedación, se pesan, los ojos están lubricados con un ungüento oftálmico estéril y se coloca en un recumbancy dorsal en un agua caliente que circula dispositivo de calentamiento bajo anestesia general, con un cono de la nariz, con isoflurano y oxígeno. En este punto, la tasa de oxígeno se reduce a 0,2 litros por minuto. </li><li> El abdomen se rocía con una solución de clorhexidina (diacetato de clorhexidina es rápidamente bacterialcidal y persistente) y se limpió con una gasa estéril tres veces o hasta que el sitio quirúrgico está limpio. </li></ol> 2. Ratón Micro-quirúrgicos y procedimientos de recuperación <ol><li> Una incisión de línea media ventral (2 cm) se realiza con unas tijeras iris a través de la piel y la pared abdominal exponer el abdomen. Una solución salina estéril gasa empapada 2x2in se utiliza para reflejar los intestinos al lado izquierdo del animal. Una inmovilización del ratón se pone en la visualización lugar en que se de la vena cava inferior (VCI). </li><li> En este punto, un nivel de mantenimiento del 2% isoflurance y 100% de oxígeno de 0,2 litros / min se utiliza para el resto de la intervención quirúrgica. </li><li> Disección roma se facilita con un hisopo aplicador estéril y extra pinzas delicadas medio del iris curva de tejidos. Se debe tener cuidado en la manipulación de los tejidos del ratón para que son extremadamente frágiles. Al entrar en la cavidad abdominal, utilice una gasa estéril salina 4x4in empapado de reflejar y exteriorizar el intestino en un campo estéril, lo que permite la visualización de la vena cava inferior. </li><li> Todas las ramas de nuevo IVC, a partir de las venas renales hasta la bifurcación ilíaca, se cauterizan, mientras que las ramas laterales se ligan con sutura de prolene 7-0 no reactivo. </li><li> El IVC está separada de la aorta, inmediatamente por debajo de las venas renales. 7-0 prolene se utiliza para ligar la vena cava inferior. </li><li> El sitio de la laparotomía se cierra en forma de dos capas. Nuestro laboratorio utiliza Vicryl 5-0, una sutura sintética, en un patrón continuo para cerrar la pared abdominal. Pegamento pegamento de la piel se utiliza para cerrar la piel. No hay puntos de sutura externos para su eliminación. </li><li> Ratones después se recuperan en una jaula de ratón individual, observada después de la operación (45 minutos a 2 horas), bajo una lámpara de calentamiento (distancia mínima de 24 centímetros de distancia de la jaula), y luego regresaron a sus viviendas originales. Medicamentos para el dolor y agentes anti-inflamatorios no se puede utilizar si no interfiere con el objetivo de este estudio. Justificación científica para suspender medicamentos para el dolor, veterinarios, y la aprobación ICUCA se necesitan. Nuestro laboratorio ha descubierto que la buprenorfina, 0,05-0,1 mg / kg vía subcutánea es un analgésico excelente pre-y post-operatorios para los ratones. Además, la lidocaína en 4mg/kg (0,4 ml / kg de una solución al 1%) puede ser infundida en el sitio de la incisión, cuando analgésicos sistémicos pueden confundir los resultados experimentales. </li><li> Animales Sham de cada grupo experimental someterse a la disección más, utilizados para el abordaje quirúrgico, con excepción de la vena cava inferior o las ramas de drenaje no se ligan o cauterizan. </li><li> La eutanasia se realiza de acuerdo con las recomendaciones establecidas por la directrices de la American Veterinary Medical Association sobre la eutanasia para los roedores. </li></ol>

<ol>
	<li>Naess, I. A., Christiansen, S. C., Romundstad, P., Cannegieter, S. C., Rosendaal, F. R., Hammerstrom, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Incidence+and+mortality+of+venous+thrombosis:+a+population-based+study.">Incidence and mortality of venous thrombosis: a population-based study.</a> J Thromb Haemost. 5, 692-699 (2007).</li><li>White, R. H., Keenan, C. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+race+and+ethnicity+on+the+incidence+of+venous+thromboembolism.">Effects of race and ethnicity on the incidence of venous thromboembolism.</a> Thromb Res. 123, S11-S17 (2009).</li><li>Heit, J. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+epidemiology+of+venous+thromboembolism+in+the+community.">The epidemiology of venous thromboembolism in the community.</a> Arterioscler Thromb Vasc Biol. 28, 370-372 (2008).</li><li>Henke, P. K., Varga, A., De, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Deep+vein+thrombosis+resolution+is+modulated+by+monocyte+CXCR2-mediated+activity+in+a+mouse+model.">Deep vein thrombosis resolution is modulated by monocyte CXCR2-mediated activity in a mouse model.</a> Arterioscler Thromb Vasc Biol. 24, 1130-1137 (2004).</li><li>McDonald, A. P., Meier, T. R., Hawley, A. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Aging+is+associated+with+impaired+thrombus+resolution+in+a+mouse+model+of+stasis+induced+thrombosis.">Aging is associated with impaired thrombus resolution in a mouse model of stasis induced thrombosis.</a> Thromb Res. 125, 72-78 (2010).</li><li>Myers, D., Farris, D., Hawley, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Selectins+influence+thrombosis+in+a+mouse+model+of+experimental+deep+venous+thrombosis.">Selectins influence thrombosis in a mouse model of experimental deep venous thrombosis.</a> J Surg Res. 108, 212-221 (2002).</li><li>Myers, D. D., Hawley, A. E., Farris, D. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=P-selectin+and+leukocyte+microparticles+are+associated+with+venous+thrombogenesis.">P-selectin and leukocyte microparticles are associated with venous thrombogenesis.</a> J Vasc Surg. 38, 1075-1089 (2003).</li><li>Myers, D. D. J. r., Rectenwald, J. E., Bedard, P. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Decreased+venous+thrombosis+with+an+oral+inhibitor+of+P+selectin.">Decreased venous thrombosis with an oral inhibitor of P selectin.</a> J Vasc Surg. 42, 329-336 (2005).</li></ol>

El modelo de ratón completa estasis de la vena cava inferior trombosis Cava se puede realizar en múltiples cepas de ratones de diferentes edades. El modelo más adaptado a los ratones de al menos 20 gramos de peso. Este modelo animal de trombosis venosa permite a los investigadores para estudiar las interacciones entre inflamatoria en la pared venosa y la trombosis venosa oclusiva. Desde nuestra experiencia, hemos encontrado que más de 10 animales por grupo son necesarios para obtener la significación estadística e...

<table border="1"><tbody><tr><td> Suministros y Equipos </td></tr><tr><td> Gas isoflurano </td></tr><tr><td> Prolene 7-0 sutura </td></tr><tr><td> Vicryl 5-0 sutura </td></tr><tr><td> La piel de cola </td></tr><tr><td> Ojo de lubricación </td></tr><tr><td> Nolvasan solución o la solución bacteriostática </td></tr><tr><td> Estereomicroscopio alcance </td></tr><tr><td> MediCapture </td></tr><tr><td> Los aplicadores de algodón </td></tr><tr><td> Gasa 2x2 </td></tr><tr><td> Almohadilla eléctrica </td></tr><tr><td> Anestesia de la máquina </td></tr><tr><td> Cuentas de vidrio instrumento esterilizador </td></tr><tr><td> Micro instrumentos quirúrgicos </td></tr><tr><td> Inmovilizadores abdominal </td></tr></tbody></table>

mouse complete stasis model of inferior vena cava thrombosis

El tromboembolismo venoso (TEV), incluye la trombosis venosa profunda (TVP) y embolismo pulmonar (EP). En los Estados Unidos (EE.UU.), la morbilidad y mortalidad elevadas tasas de TEV hacer un serio problema de salud 1.2. Después de la enfermedad cardíaca y accidente cerebrovascular, tromboembolismo venoso es la tercera enfermedad vascular tres más comunes. Sólo en los EE.UU., hay unas 900.000 personas sufren cada año, con 300.000 muertes anuales 3. Un confiable en un modelo animal in vivo para estudiar los mecanismos de esta enfermedad es necesario. Las ventajas de utilizar el modelo de ratón completa estasis de la vena cava inferior trombosis cava son varias. El modelo de ratón permite la administración de volúmenes muy pequeños de la limitada disponibilidad de los agentes de la prueba, lo que reduce drásticamente los costes. Más prometedora es la posibilidad de que los ratones con knock-outs genéticos que permiten a funciones específicas de los factores inflamatorios y de coagulación para ser delineadas. Actuales técnicas moleculares permiten la cuantificación de la pared de la vena, la sangre de trombos, todo, y el plasma para los ensayos. Sin embargo, una preocupación importante participación de este modelo es el tamaño de las limitaciones operativas y la friabilidad de los vasos. Además, debido al peso de la muestra pequeños IVC (media de 0.005 gramos) es necesario aumentar el número de animales para el análisis estadístico preciso para los tejidos, trombosis, y los ensayos de la sangre, como la reacción en cadena de la polimerasa (RT-PCR), western blot, ligado a enzimas (ELISA), zymography, la pared venosa y el análisis celular de trombos, y la sangre entera y plasma de los ensayos 4-8. La principal desventaja con el modelo de estancamiento es que la falta de flujo sanguíneo inhibe el efecto máximo de agentes terapéuticos administrados sistémica en el trombo y la pared de la vena.

Watch this Scientific Journal Video about Mouse Complete Stasis Model of Inferior Vena Cava Thrombosis at JoVE.com

Mouse Complete Stasis Model of Inferior Vena Cava Thrombosis

El modelo de ratón completa estasis de la vena cava inferior trombosis cava produce cantidades cuantificables de los tejidos de la pared venosa y trombosis. Que ha demostrado ser útil para evaluar las interacciones entre la pared de la vena y el trombo oclusivo y en la evaluación de la progresión de la aguda a la inflamación crónica.

Ratón completo modelo de estasis Inferior Vena Cava trombosis

Venous thromboembolism (VTE) includes both deep vein thrombosis (DVT) and pulmonary embolism (PE).  In the United States (U.S.), the high morbidity and ...

mouse-complete-stasis-model-of-inferior-vena-cava-thrombosis

Research

JoVE Journal

Medicine

18.1K vistas.  University of Michigan. El modelo de ratón completa estasis de la vena cava inferior trombosis cava produce cantidades cuantificables de los tejidos de la pared venosa y trombosis. Que ha demostrado ser útil para evaluar las interacciones entre la pared de la vena y el trombo oclusivo y en la evaluación de la progresión de la aguda a la inflamación crónica.

Video: Mouse Complete Stasis Model of Inferior Vena Cava Thrombosis

Electrolytic Inferior Vena Cava Model (EIM) of Venous Thrombosis

Animal models serve a vital role in deep venous thrombosis (DVT) research in order to study thrombus formation, thrombus resolution and to test potential therapeutic compounds (1). New compounds to be utilized in the treatment and prevention of DVT are currently being developed. The delivery of potential therapeutic antagonist compounds to an affected thrombosed vein has been problematic. In the context of therapeutic applications, a model that uses partial stasis and consistently generates thrombi within a major vein has been recently established. The Electrolytic Inferior vena cava Model (EIM) is mouse model of DVT that permits thrombus formation in the presence of continuous blood flow. This model allows therapeutic agents to be in contact with the thrombus in a dynamic fashion, and is more sensitive than other models of DVT (1). In addition, this thrombosis model closely simulates clinical situations of thrombus formation and is ideal to study venous endothelial cell activation, leukocyte migration, venous thrombogenesis, and to test therapeutic applications (1). The EIM model is technically simple, easily reproducible, creates consistent thrombi sizes and allows for a large sample (i.e. thrombus and vein wall) which is required for analytical purposes.

Electrolytic Inferior Vena Cava Model EIM of Venous Thrombosis

The electrolytic induction of endothelial activation to the internal surface of the Inferior Vena Cava results in venous type thrombus formation due to endothelial activation and partial blood stasis, two components of Virchow's triad.

Electrolítico de la vena cava inferior del modelo (EIM) de la Trombosis Venosa

Animal models serve a vital role in deep venous thrombosis (DVT) research in order to study thrombus formation, thrombus resolution and to test potential ...

Investigación

Medicina

Quantitative Analysis and Characterization of Atherosclerotic Lesions in the Murine Aortic Sinus

Atherosclerosis is a disease of the large arteries and a major underlying cause of myocardial infarction and stroke. Several different mouse models have been developed to facilitate the study of the molecular and cellular pathophysiology of this disease. In this manuscript we describe specific techniques for the quantification and characterization of atherosclerotic lesions in the murine aortic sinus and ascending aorta. The advantage of this procedure is that it provides an accurate measurement of the cross-sectional area and total volume of the lesion, which can be used to compare atherosclerotic progression across different treatment groups. This is possible through the use of the valve leaflets as an anatomical landmark, together with careful adjustment of the sectioning angle. We also describe basic staining methods that can be used to begin to characterize atherosclerotic progression. These can be further modified to investigate antigens of specific interest to the researcher. The described techniques are generally applicable to a wide variety of existing and newly created dietary and genetically-induced models of atherogenesis.

We describe procedures to quantify and characterize atherosclerotic lesions in mouse models using precision sectioning of the aortic sinus and ascending aorta combined with histochemical and immunohistochemical analysis.

Atherosclerosis is a disease of the large arteries and a major underlying cause of myocardial infarction and stroke. Several different mouse models have ...

Implantation of Inferior Vena Cava Interposition Graft in Mouse Model

Biodegradable scaffolds seeded with bone marrow mononuclear cells (BMCs) are often used for reconstructive surgery to treat congenital cardiac anomalies. The long-term clinical results showed excellent patency rates, however, with significant incidence of stenosis. To investigate the cellular and molecular mechanisms of vascular neotissue formation and prevent stenosis development in tissue engineered vascular grafts (TEVGs), we developed a mouse model of the graft with approximately 1 mm internal diameter. First, the TEVGs were assembled from biodegradable tubular scaffolds fabricated from a polyglycolic acid nonwoven felt mesh coated with &#949;-caprolactone and L-lactide copolymer. The scaffolds were then placed in a lyophilizer, vacuumed for 24 hr, and stored in a desiccator until cell seeding. Second, bone marrow was collected from donor mice and mononuclear cells were isolated by density gradient centrifugation. Third, approximately one million cells were seeded on a scaffold and incubated O/N. Finally, the seeded scaffolds were then implanted as infrarenal vena cava interposition grafts in C57BL/6 mice. The implanted grafts demonstrated excellent patency (&#62;90%) without evidence of thromboembolic complications or aneurysmal formation. This murine model will aid us in understanding and quantifying the cellular and molecular mechanisms of neotissue formation in the TEVG.

To improve our knowledge of cellular and molecular neotissue formation, a murine model of the TEVG was recently developed. The grafts were implanted as infrarenal vena cava interposition grafts in C57BL/6 mice. This model achieves similar results to those achieved in our clinical investigation, but over a far shortened time-course.

Implantación de la vena cava inferior interposición del injerto en modelo de ratón

Biodegradable scaffolds seeded with bone marrow mononuclear cells (BMCs) are often used for reconstructive surgery to treat congenital cardiac anomalies. ...

Orthotopic Injection of Breast Cancer Cells into the Mammary Fat Pad of Mice to Study Tumor Growth.

Breast cancer growth can be studied in mice using a plethora of models. Genetic manipulation of breast cancer cells may provide insights into the functions of proteins involved in oncogenic progression or help to discover new tumor suppressors. In addition, injecting cancer cells into mice with different genotypes might provide a better understanding of the importance of the stromal compartment. Many models may be useful to investigate certain aspects of disease progression but do not recapitulate the entire cancerous process. In contrast, breast cancer cells engraftment to the mammary fat pad of mice better recapitulates the location of the disease and presence of the proper stromal compartment and therefore better mimics human cancerous disease. In this article, we describe how to implant breast cancer cells into mice orthotopically and explain how to collect tissues to analyse the tumor milieu and metastasis to distant organs. Using this model, many aspects (growth, angiogenesis, and metastasis) of cancer can be investigated simply by providing a proper environment for tumor cells to grow.

Cancer is a complex disease that is influenced by the tissue surrounding the tumor as well as local pro- and anti-inflammatory mediators. Therefore, orthotropic injection models, rather than subcutaneous models may be useful to study cancer progression in a manner that better mimics human pathology.

Inyección ortotópico de células de cáncer de mama en el Mamaria Fat Pad de ratones para estudiar el crecimiento del tumor.

Breast cancer growth can be studied in mice using a plethora of models. Genetic manipulation of breast cancer cells may provide insights into the ...

Ferric Chloride-induced Thrombosis Mouse Model on Carotid Artery and Mesentery Vessel

Severe thrombosis and its ischemic consequences such as myocardial infarction, pulmonary embolism and stroke are major worldwide health issues. The ferric chloride injury is now a well-established technique to rapidly and accurately induce the formation of thrombi in exposed veins or artery of small and large diameter. This model has played a key role in the study of the pathophysiology of thrombosis, in the discovery and validation of novel antithrombotic drugs and in the understanding of the mechanism of action of these new agents. Here, the implementation of this technique on a mesenteric vessel and carotid artery in mice is presented. The method describes how to label circulating leukocytes and platelets with a fluorescent dye and to observe, by intravital microscopy on the exposed mesentery, their accumulation at the injured vessel wall which leads to the formation of a thrombus. On the carotid artery, the occlusion caused by the clot formation is measured by monitoring the blood flow with a Doppler probe.

The FeCl3 induced thrombosis model in mice is described herein. A method to monitor thrombus growth by intravital microscopy observation on a mesenteric vessel and by blood flow measurement in the carotid artery is presented.

Trombosis modelo de ratón de la arteria carótida y Mesenterio Embarcaciones inducida por cloruro férrico-

Severe thrombosis and its ischemic consequences such as myocardial infarction, pulmonary embolism and stroke are major worldwide health issues. The ferric ...

Ferric Chloride-induced Murine Thrombosis Models

Arterial thrombosis (blood clot) is a common complication of many systemic diseases associated with chronic inflammation, including atherosclerosis, diabetes, obesity, cancer and chronic autoimmune rheumatologic disorders. Thrombi are the cause of most heart attacks, strokes and extremity loss, making thrombosis an extremely important public health problem. Since these thrombi stem from inappropriate platelet activation and subsequent coagulation, targeting these systems therapeutically has important clinical significance for developing safer treatments. Due to the complexities of the hemostatic system, in vitro experiments cannot replicate the blood-to-vessel wall interactions; therefore, in vivo studies are critical to understand pathological mechanisms of thrombus formation. To this end, various thrombosis models have been developed in mice. Among them, ferric chloride (FeCl3) induced vascular injury is a widely used model of occlusive thrombosis that reports platelet activation and aggregation in the context of an aseptic closed vascular system. This model is based on redox-induced endothelial cell injury, which is simple and sensitive to both anticoagulant and anti-platelets drugs. The time required for the development of a thrombus that occludes blood flow gives a quantitative measure of vascular injury, platelet activation and aggregation that is relevant to thrombotic diseases. We have significantly refined this FeCl3-induced vascular thrombosis model, which makes the data highly reproducible with minimal variation. Here we describe the model and present representative data from several experimental set-ups that demonstrate the utility of this model in thrombosis research.

We report a refined procedure of the ferric chloride (FeCl3)-induced thrombosis models on carotid and mesenteric artery as well as vein, characterized efficiently using intravital microscopy to monitor time to occlusive thrombi formation.

Férricos modelos murinos trombosis inducida por cloruro

Arterial thrombosis (blood clot) is a common complication of many systemic diseases associated with chronic inflammation, including atherosclerosis, ...

Patch Angioplasty in the Rat Aorta or Inferior Vena Cava

Pericardial patches are commonly used in vascular surgery to close vessels. To facilitate studies of the neointimal hyperplasia that forms on the patch, we developed a rat model of patch angioplasty that can be used in either a vein or an artery, creating a patch venoplasty or a patch arterioplasty, respectively. Technical aspects of this model are discussed. The infra-renal IVC or aorta are dissected and then clamped proximally and distally. A 3 mm venotomy or arteriotomy is performed in the infrarenal inferior vena cava or aorta of 6 to 8 week-old Wistar rats. A bovine pericardial patch (3 mm x 1.5 mm x 0.6 mm) is then used to close the site using a 10-0 nylon suture. Compared to arterial patches, venous patches show increased neointimal thickness on postoperative day 7. This novel model of pericardial patch angioplasty can be used to examine neointimal hyperplasia on vascular biomaterials, as well as to compare the differences between the arterial and venous environments.

We have established a model of pericardial patch angioplasty that can be used in either small-diameter veins or arteries. This model can be used to compare venous and arterial neointimal hyperplasia formation.

Parche angioplastia en la aorta de rata o de la vena cava inferior

Pericardial patches are commonly used in vascular surgery to close vessels. To facilitate studies of the neointimal hyperplasia that forms on the patch, ...

A Rat Model of Orthotopic Liver Transplantation Using a Novel Magnetic Anastomosis Technique for Suprahepatic Vena Cava Reconstruction

The rat model of orthotopic liver transplantation (OLT) is essential for transplant research. It is a very sophisticated animal model and requires a steep learning curve. The introduction of the cuff technique for anastomosis of the portal vein (PV) and infrahepatic vena cava (IHVC) has significantly simplified the transplant procedure in rats. However, due to the short anterior wall of the recipients' suprahepatic vena cava (SHVC), the cuff technique is very difficult to use for the reconstruction of the SHVC. Most researchers in this field still use the hand-suture technique for SHVC reconstruction, which makes it the bottleneck step in rat orthotopic liver transplantation. The magnetic anastomosis technique (i.e., magnamosis) is a method of connecting two vessels using the attractive force between two magnets. Our recent study has shown that the magnetic anastomosis technique is superior to the hand-suture technique for SHVC reconstruction in rats. In this article, we show a step-by-step protocol for SHVC reconstruction in rats using the novel magnetic anastomosis technique. In this model, the reconstruction of the PV and IHVC was performed by the standard cuff technique, while the reconstruction of the bile duct (BD) was performed by a stent technique. The hepatic re-arterialization was not performed. The magnetic anastomosis technique made SHVC reconstruction much easier and significantly shortened the anphepatic phase. After a reasonable learning curve, even researchers without advanced microsurgical skills can produce reliable and reproducible results using this rat model of OLT.

The reconstruction of the suprahepatic vena cava (SHVC) remains a difficult step in rat orthotopic liver transplantation. In this article, we show a step-by-step protocol for SHVC reconstruction in rats using a novel magnetic anastomosis technique.

Un modelo de rata de Orthotopic trasplante del hígado usando una técnica de Anastomosis magnética novela para la reconstrucción de la Vena Cava suprahepática

The rat model of orthotopic liver transplantation (OLT) is essential for transplant research. It is a very sophisticated animal model and requires a steep ...

Ferric Chloride-induced Canine Carotid Artery Thrombosis: A Large Animal Model of Vascular Injury

Occlusive arterial thrombosis leading to cerebral ischemic stroke and myocardial infarction contributes to ~13 million deaths every year globally. Here, we have translated a vascular injury model from a small animal into a large animal (canine), with slight modifications that can be used for pre-clinical screening of prophylactic and thrombolytic agents. In addition to the surgical methods, the modified protocol describes the step-by-step methods to assess carotid artery canalization by angiography, detailed instructions to process both the brain and carotid artery for histological analysis to verify carotid canalization and cerebral hemorrhage, and specific parameters to complete an assessment of downstream thromboembolic events by utilizing magnetic resonance imaging (MRI). In addition, specific procedural changes from the previously well-established small animal model necessary to translate into a large animal (canine) vascular injury are discussed.

Here, we present the modifications necessary to a well characterized and commonly used small animal ferric chloride-induced (FeCl3) carotid artery injury model for use in a large animal vascular injury model. The resulting model can be utilized for pre-clinical trial assessment of both prophylactic and thrombolytic pharmacological and mechanical interventions.

Trombosis de la arteria carótida canina cloruro férrico-inducida: Un modelo Animal grande de lesión Vascular

Occlusive arterial thrombosis leading to cerebral ischemic stroke and myocardial infarction contributes to ~13 million deaths every year globally. Here, ...

Isometric Contractility Measurement of the Mouse Mesenteric Artery Using Wire Myography

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and drugs. It is widely used in many studies on the physiological functions of vascular smooth muscle, the pathogenesis of vascular diseases such as hypertension, and the development of smooth muscle relaxant drugs. The mouse is a widely used model animal with a large pool of disease models and genetically modified strains. We introduced this method to measure the isometric contraction of mouse mesenteric artery in detail. A 1.4-mm segment of mouse mesenteric resistance artery was isolated and mounted on a myograph chamber by passing two steel wires through its lumen. After equilibration and normalization steps, the vessel segment was potentiated by a high-K+ solution twice prior to the contraction assay. As an example of the application of this method in drug development, we measured the relaxant effect of a novel natural substance, neoliensinine, isolated from a Chinese herb, embryos of the lotus seed (Nelumbo nucifera Gaertn.) on mouse mesenteric arteries. The vessel segments mounted on the myograph chamber were stimulated with a high-K+ solution. When the force tension reached a stable sustained phase, cumulative doses of neoliensinine were added to the chamber. We found that neoliensinine had a dose-dependent relaxant effect on smooth muscle contraction, thus suggesting that it bears potential activity against hypertension. In addition, as the vessel segment can survive at least 4 hours after mounting and maintain contractility induced by the high-K+ solution for many times, we suggest that the wire myograph system may be used for the time-consuming process of drug screening.

The wire myograph technique is used to investigate vascular smooth muscle functions and screen new drugs. We report a detailed protocol for measuring the isometric contractility of the mouse mesenteric artery and for screening new relaxants of vascular smooth muscle.

Medición de la contractilidad isométrica de la arteria mesentérica de ratón con cable miografía

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and ...