This work was supported by a Wisconsin Partnership Program New Investigator Award (ID 2832), a National Heart, Lung, Blood Institute R01 Grant (HL-068673) and a T32 training Grant (HL-110853). We thank Dr. Melina Kibbe&#8217;s group at Northwestern University for providing helpful information.

Declaración de Ética: NOTA: Todos los procedimientos se ajustan a la Guía para el Cuidado y Uso de Animales de Laboratorio (National Academies Press, 8ª edición, 2011), y los protocolos aprobados por el Comité de Cuidado y Uso de Animales Institucional de la Universidad de Wisconsin. Todas las cirugías se realizaron bajo anestesia con isoflurano (a través de la inhalación, caudal de 2 ml / min), y se hicieron todos los esfuerzos para minimizar el sufrimiento. Los animales fueron sacrificados en una cámara llena gradualmente con CO 2. 1. La inducción de la anestesia general y la incisión de la piel <ol><li> Utilice macho C57BL / 6 ratones con una edad de 12-16 semanas y el peso de 23-28 g. Anestesiar al animal mediante isoflurano a través de un aparato de anestesia de circuito cerrado. Confirmar anestesia adecuada apretando su dedo del pie. Aplique un poco de ungüento de lágrimas artificiales en los ojos para evitar la sequedad. </li><li> Coloque el ratón en una posición supina con la extremidad posterior izquierda ligeramente secuestrado y la articulación de la rodilla slightly flexionada. Afeitarse el pelo alrededor de la región superior del muslo izquierdo y desinfectar la piel con clorhexidina, aplicado tres veces con algodón estéril aplicadores con punta. Utilice un esterilizador de cuentas caliente para mantener los instrumentos esterilizados durante la cirugía. </li><li> Coloque un paño de gasa esterilizada sobre la pierna izquierda. Hacer una incisión longitudinal en la piel 1,5 cm recta en el muslo medial izquierdo de la articulación de la rodilla hasta la ingle (Figura 1A). </li></ol> 2. La exposición de la arteria femoral y Ramas NOTA: La Figura 1B muestra la anatomía vascular grave en la parte superior del muslo. La arteria femoral superficial (SFA) se puede observar a través de un microscopio de disección cuando la incisión de la piel se hace adecuadamente a lo largo de la línea media del muslo medial. <ol><li> Abre la fascia delgada que recubre el SFA con finas pinzas y luego bucle del SFA con 9-0 sutura de nylon. NOTA: En este proceso, el nervio femoral es fácil de poner comoide, pero la vena femoral adyacente requiere mucha atención, porque su pared es tan delgada que perfora y sangra fácilmente. La técnica clave es centrarse en la propia arteria; siempre que la disección se lleva a cabo cerca de la arteria, cualquier daño accidental vena es evitable. </li><li> Una vez que la SFA se enlaza, mantenga la sutura con una pinza de mosquito y tire de él ligeramente hacia abajo. Esta maniobra hace más disección más fácil, pero para evitar daños en la arteria no tire con demasiada fuerza. </li><li> Continuar la disección de la arteria proximal para exponer y bucle de la arteria femoral común (CFA) con 9-0 sutura de nylon. NOTA: Por lo general, rizando el Comité de Libertad Sindical en un punto distal a la arteria epigástrica (Figura 1B) es suficiente para la inserción de cables. Mantenga el lazo de sutura CFA con pinzas de mosquitos de la misma manera. </li><li> Tire del lazo de sutura CFA hacia arriba y el lazo de sutura SFA hacia abajo para destapar la bifurcación de SFA y la arteria femoral profunda (DFA). La DFA, la rama principal del Comité de Libertad Sindical,va hacia el músculo del muslo (Figura 1B). NOTA: El cable se inserta desde la DFA después. </li><li> Pase el DFA con mucha precaución debido a que la vena adyacente está enredado por debajo de la arteria. En caso de sangrado de la vena, un 30 a 60 seg compresión simple con una gasa es generalmente suficiente para producir hemostasis. </li><li> Una vez que el DFA se enlaza con 9-0 sutura de nylon, asegúrese de que los tres bucles de sutura están en sus posiciones correctas (Figura 2A). </li><li> Proceder con la disección de la DFA distalmente para ligar la arteria en un punto lo más lejos posible de la bifurcación. Cortar sutura extra, pero dejar una cierta longitud de manera que los dos extremos se pueden celebrar con pinzas de mosquito, que estabilizan la DFA hacer la inserción de cables más fácil. </li></ol> 3. Creación de Lesión alambre <ol><li> Inserte el cable a través de DFA <ol><li> Colar las suturas CFA, SFA, y DFA para establecer una pinza hemostática temporal. </li><li> Haz un pequeño agujero en el DFA con microtijeras. Insertar las puntas de los micropinzas curvadas para dilatar el agujero suficientemente grande para un alambre con un diámetro de 0,015 pulgadas (Figura 2B demuestra la diferencia de tamaño entre la arteria y el cable). <ol><li> Antes de hacer un agujero en la DFA, aflojar el lazo de sutura CFA y suavemente sujetar / apriete el DFA varias veces con micropinzas-opacos punta. Esta maniobra se amplía el diámetro arterial notablemente para facilitar la inserción de cables. Humedecer el alambre a fondo con PBS también es importante para una inserción suave. </li><li> Al hacer un agujero en la DFA, no recortar la pared arterial, pero hacer un pequeño pellizco en forma de V y pellizcar la solapa con micropinzas de punta afilada para la manipulación. </li></ol></li><li> Inserte el cable por el agujero adecuadamente dilatado en la DFA. Avanzar suavemente el alambre hacia la arteria ilíaca mientras afloja el lazo de sutura alrededor del CFA. 5-8 mm es suficiente para cubrir toda la longitud CFA en la mayoría de los casos (Figura 2C </strong&gt;). No inserte el cable de fuerza como una perforación arterial por lo general resulta en la pérdida del ratón. Acortar el cable a aproximadamente 15 mm de longitud ayuda a la manipulación. </li></ol></li><li> Deja alambre dentro de 1 Min <ol><li> Después de insertar el cable a la longitud óptima, se deja durante 1 min a estirar demasiado la pared arterial. Humedezca el alambre y las arterias con PBS para evitar que se seque. Otra sutura de nylon 9-0 se puede colocar alrededor de la DFA proximal al punto de inserción de alambre durante este tiempo de espera. Esta sutura se utilizará más adelante para cerrar el orificio de la DFA después de la extracción del alambre. </li></ol></li><li> Recuperar alambre y reanudar el flujo sanguíneo <ol><li> Sujete el cable con pinzas de canulación del vaso, y tirar de ella con suavidad; el cable debe ser recuperada sin problemas. Para superar cualquier resistencia, causada generalmente por la adhesión del alambre a la pared arterial, retroceso del cable con rotación suave es útil. Una vez más, una maniobra contundente podría resultar en la pérdida de los animales. </li><li> Laespués de recuperar el cable, ligar la DFA en el punto de bifurcación con sutura de nylon 9-0. No ligar demasiado cerca de la bifurcación, que pondría en peligro el flujo de salida de sangre a la SFA. </li><li> Aflojar los lazos de sutura SFA y CFA para reanudar el flujo de sangre normal. Tenga en cuenta que el Comité de Libertad Sindical es bien expandimos la inserción del cable (Figura 2D). </li></ol></li></ol> 4. Cierre de la piel y Cuidado Postoperatorio <ol><li> Sangra de control. Exudación Minor puede ser detenida por compresión simple; algunos importantes hemorragias arteriales requieren puntos de sutura con una sutura de nylon 11-0. Estas arterias suturadas eventualmente resultarán en la oclusión arterial. </li><li> Cierre de la piel de la herida con grapadoras de piel (generalmente 3-4 grapas). Mantenga el área de la herida esterilizado mediante la aplicación de solución quirúrgica clorhexidina. </li><li> Aplicar tratamientos post-quirúrgicos. Controlar el dolor del ratón con inyecciones subcutáneas de buprenorfina. Mantenga el ratón en la plataforma de calentamiento hasta que se haya recuperado sufciente conciencia de mantener decúbito esternal. Casa en una jaula limpia y sin compañía de otros animales hasta que se recuperó totalmente. </li></ol>

<ol>
	<li>Dzau, V. J., Braun-Dullaeus, R. C., Sedding, D. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Vascular+proliferation+and+atherosclerosis:+new+perspectives+and+therapeutic+strategies.">Vascular proliferation and atherosclerosis: new perspectives and therapeutic strategies.</a> Nat Med. 8 (11), 1249-1256 (2002).</li><li>Hui, D. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intimal+hyperplasia+in+murine+models.">Intimal hyperplasia in murine models.</a> Curr Drug Targets. 9 (3), 251-260 (2008).</li><li>Ferns, G. A., Avades, T. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+mechanisms+of+coronary+restenosis:+insights+from+experimental+models.">The mechanisms of coronary restenosis: insights from experimental models.</a> Int J Exp Pathol. 81 (2), 63-88 (2000).</li><li>Sata, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+mouse+model+of+vascular+injury+that+induces+rapid+onset+of+medial+cell+apoptosis+followed+by+reproducible+neointimal+hyperplasia.">A mouse model of vascular injury that induces rapid onset of medial cell apoptosis followed by reproducible neointimal hyperplasia.</a> J Mol Cell Cardiol. 32 (11), 2097-2104 (2000).</li><li>Xu, Q. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mouse+models+of+arteriosclerosis:+from+arterial+injuries+to+vascular+grafts.">Mouse models of arteriosclerosis: from arterial injuries to vascular grafts.</a> Am J Pathol. 165 (1), 1-10 (2004).</li><li>Lv, L., Meng, Q., Ye, M., Wang, P., Xue, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=STAT4+deficiency+protects+against+neointima+formation+following+arterial+injury+in+mice.">STAT4 deficiency protects against neointima formation following arterial injury in mice.</a> J Mol Cell Cardiol. 74C, 284-294 (2014).</li><li>Zhu, B., Kuhel, D. G., Witte, D. P., Hui, D. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Apolipoprotein+E+inhibits+neointimal+hyperplasia+after+arterial+injury+in+mice.">Apolipoprotein E inhibits neointimal hyperplasia after arterial injury in mice.</a> Am J Pathol. 157 (6), 1839-1848 (2000).</li><li>Fu, Y., Duru, E. A., Davies, M. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+metabolic+syndrome+on+the+response+to+arterial+injury.">Effect of metabolic syndrome on the response to arterial injury.</a> J Surg Res. , (2014).</li><li>Kuhel, D. G., Zhu, B., Witte, D. P., Hui, D. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Distinction+in+genetic+determinants+for+injury-induced+neointimal+hyperplasia+and+diet-induced+atherosclerosis+in+inbred+mice.">Distinction in genetic determinants for injury-induced neointimal hyperplasia and diet-induced atherosclerosis in inbred mice.</a> Arterioscler Thromb Vasc Biol. 22 (6), 955-960 (2002).</li><li>Cooley, B. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mouse+strain+differential+neointimal+response+in+vein+grafts+and+wire-injured+arteries.">Mouse strain differential neointimal response in vein grafts and wire-injured arteries.</a> Circ J. 71 (10), 1649-1652 (2007).</li><li>Ishigami, N., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Deficiency+of+CuZn+superoxide+dismutase+promotes+inflammation+and+alters+medial+structure+following+vascular+injury.">Deficiency of CuZn superoxide dismutase promotes inflammation and alters medial structure following vascular injury.</a> J Atheroscler Thromb. 18 (11), 1009-1017 (2011).</li></ol>

The authors declare no competing financial interests.

El procedimiento lesión alambre es aplicable a todas las cepas de ratones, siempre y cuando sus estructuras anatómicas son similares. 4 En este trabajo, hemos utilizado macho C57BL / 6 ratones con una edad de 12 a 16 semanas. Como hemos aprendido de nuestros estudios preliminares, las arterias femorales de ratones más jóvenes de 10 semanas son a menudo tan pequeña que la inserción del alambre es todo un reto. Por otro lado, la inserción del alambre en ratones de más de 16 semanas es técnicamente más...

In the era of expanded application of endovascular treatments for various cardiovascular diseases, restenosis after angioplasty is one of the major problems for patients undergoing such treatments. Damage to the vascular endothelium at the time of angioplasty, in concert with the atherosclerotic background, induces excessive smooth muscle cell proliferation in the medial layer, resulting in neointimal hyperplasia.1,2 A viable animal model that recapitulates post-angioplasty neointimal hyperplasia is, thus, important not only for the investigation of disease mechanisms but also for the development of effective therapeutics to treat this pathology.
Mice represent an excellent model animal to recapitulate neointimal hyperplasia for the following reasons: the genetic backgrounds of experimental mice are well established; a wide variety of genetically-modified strains are available;2 obtaining littermates of the same background is easy; and the cost of the animals is relatively low. Arterial ligation model and wire injury model are the two most common mouse models of mechanically-induced neointimal hyperplasia. The arterial ligation model is easy to create, but physiologically dissimilar to the actual angioplasty procedure. The wire injury model closely mimics actual angioplasty procedures but is technically difficult due to the small size of mouse arteries.2,3 Sata et al. first described a wire injury method for mouse femoral arteries based on the anatomical structure of the vasculature and the use of proper-sized flexible wire. Utilizing this technique, they succeeded in reproducibly inducing neointimal hyperplasia in various strains of mice.4
Although femoral wire injury is a well-established model, some of the technical aspects of the technique are highly challenging compared to other models such as ligation.5 The purpose of this paper is to describe our mouse wire injury model procedures in detail, which is a modified version of Sata&#8217;s original method. We have made two main modifications: 1) Looping only the arteries, and 2) No lidocaine use.

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			<td height="21" style="height:21px;width:216px;">Dilation Forceps</td>
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a murine model of arterial restenosis: technical aspects of femoral wire injury

La enfermedad cardiovascular causada por la aterosclerosis es la causa principal de muerte en el mundo desarrollado. El estrechamiento de la luz del vaso, debido al desarrollo de la placa aterosclerótica o la ruptura de placas establecidos, interrumpe el flujo normal de la sangre que conduce a diversas morbilidades, tales como infarto de miocardio y accidente cerebrovascular. En los procedimientos endovasculares clínicas como la angioplastia se realizan comúnmente para reabrir el lumen. Sin embargo, estos tratamientos dañan inevitablemente la pared del vaso, así como el endotelio vascular, la activación de una respuesta de cicatrización excesiva y el desarrollo de una placa neointimal que se extiende en el lumen del vaso que causan la reestenosis (re-estrechamiento). La restenosis sigue siendo una causa importante de fracaso de los tratamientos endovasculares para la aterosclerosis. Por lo tanto, los modelos animales preclínicos de reestenosis son de vital importancia para la investigación de los mecanismos fisiopatológicos, así como de la traducción se acerca a las intervenciones vasculares. Entre varios EXPERIME murinomodelos NTAL, lesiones alambre arteria femoral es ampliamente aceptado como el más adecuado para estudios de reestenosis post-angioplastia porque se parece mucho al procedimiento de angioplastia que daña tanto el endotelio y la pared del vaso. Sin embargo, muchos investigadores tienen dificultades para la utilización de este modelo debido a su alto grado de dificultad técnica. Esto es principalmente porque un alambre de metal tiene que ser insertado en la arteria femoral, que es aproximadamente tres veces más delgado que el alambre, para generar suficiente para inducir la lesión neoíntima prominente. A continuación, describimos los detalles quirúrgicos esenciales para superar de manera efectiva las principales dificultades técnicas de este modelo. Siguiendo los procedimientos presentados, la realización de la lesión de alambre de la arteria femoral del ratón se vuelve más fácil. Una vez familiarizado, todo el procedimiento puede ser completado dentro de 20 min.

Cuatro semanas después de la cirugía, la cosecha de la arteria femoral alambre heridos y hacen muestra de bloque una parafina para analizar la formación de la neoíntima. Inducir la anestesia general como se describió anteriormente. Abra el pecho inmediatamente e inserte una mariposa aguja de calibre 20 al ventrículo izquierdo. Conecte la línea de una botella de infusión. Realizar fijación por perfusión a través de infusión por goteo de PBS seguido de paraformaldehído al 4% (por lo general 10 a 20 m...

Watch this Scientific Journal Video about A Murine Model of Arterial Restenosis: Technical Aspects of Femoral Wire Injury at JoVE.com

A Murine Model of Arterial Restenosis: Technical Aspects of Femoral Wire Injury

El modelo de lesión de alambre arteria femoral de ratón de la reestenosis es técnicamente difícil. En este protocolo se muestran los detalles técnicos clave esencial para llevar a cabo con éxito la lesión de alambre para inducir neointima consistente para estudios de reestenosis.

Un modelo murino de restenosis arterial: Aspectos Técnicos del femoral Lesiones alambre

Cardiovascular disease caused by atherosclerosis is the leading cause of death in the developed world. Narrowing of the vessel lumen, due to ...

a-murine-model-arterial-restenosis-technical-aspects-femoral-wire

Research

JoVE Journal

Medicine

19.9K vistas.  University of Wisconsin School of Medicine and Public Health.  El modelo de lesión de alambre arteria femoral de ratón de la reestenosis es técnicamente difícil. En este protocolo se muestran los detalles técnicos clave esencial para llevar a cabo con éxito la lesión de alambre para inducir neointima consistente para estudios de reestenosis. 

Video: A Murine Model of Arterial Restenosis: Technical Aspects of Femoral Wire Injury

Microsurgical Venous Pouch Arterial-Bifurcation Aneurysms in the Rabbit Model: Technical Aspects

For ruptured human cerebral aneurysms endovascular embolization has become an equivalent alternative to aneurysm clipping.1 However, large clinical trials have shown disappointing long-term results with unacceptable high rates of aneurysm recanalization and delayed aneurysm rupture.2 To overcome these problems, animal experimental studies are crucial for the development of better endovascular devices.3-5
Several animal models in rats, rabbits, canines and swine are available.6-8 Comparisons of the different animal models showed the superiority of the rabbit model with regard to hemodynamics and comparability of the coagulation system and cost-effectiveness.9-11 
The venous pouch arterial bifurcation model in rabbits is formed by a venous pouch sutured into an artificially created true bifurcation of both common carotid arteries (CCA). The main advantage of this model are true bifurcational hemodynamics.12 The major drawbacks are the sofar high microsurgical technical demands and high morbidity and mortality rates of up to 50%.13 These limitations have resulted in less frequent use of this aneurysm model in the recent years. These shortcomings could be overcome with improved surgical procedures and modified peri- and postoperative analgetic management and anticoagulation.14-16 Our techniques reported in this paper demonstrate this optimized technique for microsurgical creation of arterial bifurcation aneurysms.

An optimized technique for the microsurgical creation of arterial bifurcation aneurysms mimicking bifurcation human cerebral aneurysms is described. A venous pouch is sutured into an artificially created true bifurcation of both common carotid arteries. Facilitated microsurgical techniques and aggressive postoperative anticoagulation and analgesia lead to minimized morbidity rates and high aneurysm patency rates.

Microcirugía venosa bolsa arterial-Bifurcación Los aneurismas en el modelo de conejo: Aspectos Técnicos

For ruptured human cerebral aneurysms endovascular embolization has become an equivalent alternative to aneurysm clipping.1 However, large clinical trials ...

Investigación

Medicina

Femoral Arterial and Venous Catheterization for Blood Sampling, Drug Administration and Conscious Blood Pressure and Heart Rate Measurements

In multiple fields of study, access to the circulatory system in laboratory studies is necessary. Pharmacological studies in rats using chronically implanted catheters permit a researcher to effectively and humanely administer substances, perform repeated blood sampling and assists in conscious direct measurements of blood pressure and heart rate. Once the catheter is implanted long-term sampling is possible. Patency and catheter life depends on multiple factors including the lock solution used, flushing regimen and catheter material. This video will demonstrate the methodology of femoral artery and venous catheterization of the rat. In addition the video will demonstrate the use of the femoral venous and arterial catheters for blood sampling, drug administration and use of the arterial catheter in taking measurements of blood pressure and heart rate in a conscious freely-moving rat. A tether and harness attached to a swivel system will allow the animal to be housed and have samples taken by the researcher with minimal disruption to the animal. To maintain patency of the catheter, careful daily maintenance of the catheter is required using lock solution (100 U/ml heparinized saline), machine-ground blunt tip syringe needles and the use of syringe filters to minimize potential contamination. With careful aseptic surgical techniques, proper catheter materials and careful catheter maintenance techniques, it is possible to sustain patent catheters and healthy animals for long periods of time (several weeks).

Chronic catheterization of blood vessels in the rat is often required for administration of substances, obtain blood sample over a period of time or for direct conscious blood pressure measurements. Femoral arterial catheterization of the rat and corresponding measurements of blood pressure in the conscious animal will be demonstrated.

El cateterismo femoral arterial y venosa para muestras de sangre, la Administración de Drogas y la presión arterial consciente y mediciones del ritmo cardíaco

In multiple fields of study, access to the circulatory system in laboratory studies is necessary. Pharmacological studies in rats using chronically ...

A Murine Model of Stent Implantation in the Carotid Artery for the Study of Restenosis

Despite the considerable progress made in the stent development in the last decades, cardiovascular diseases remain the main cause of death in western countries. Beside the benefits offered by the development of different drug-eluting stents, the coronary revascularization bears also the life-threatening risks of in-stent thrombosis and restenosis. Research on new therapeutic strategies is impaired by the lack of appropriate methods to study stent implantation and restenosis processes. Here, we describe a rapid and accessible procedure of stent implantation in mouse carotid artery, which offers the possibility to study in a convenient way the molecular mechanisms of vessel remodeling and the effects of different drug coatings.

A model of stent implantation in mouse carotid artery is described. Compared to other similar methods, this procedure is very rapid, simple and accessible, offering the possibility to study in a convenient way the vascular wall reaction to different drug-eluting stents and the molecular mechanisms of restenosis.

Un modelo murino de la implantación de un stent en la arteria carótida para el Estudio de la reestenosis

Despite  the considerable progress made in the stent development in the last decades,  cardiovascular diseases remain the main cause of death in western ...

Technical Aspects of the Mouse Aortocaval Fistula

Technical aspects of creating an arteriovenous fistula in the mouse are discussed. Under general anesthesia, an abdominal incision is made, and the aorta and inferior vena cava (IVC) are exposed. The proximal infrarenal aorta and the distal aorta are dissected for clamp placement and needle puncture, respectively. Special attention is paid to avoid dissection between the aorta and the IVC. After clamping the aorta, a 25 G needle is used to puncture both walls of the aorta into the IVC. The surrounding connective tissue is used for hemostatic compression. Successful creation of the AVF will show pulsatile arterial blood flow in the IVC. Further confirmation of successful AVF can be achieved by post-operative Doppler ultrasound.

The goal is to produce an arteriovenous fistula that is simple and reproducible. This method does not use sutures or glue adhesive. Therefore the samples can be used with the least amount of foreign materials for analysis.

Aspectos técnicos de la aortocava Fístula Ratón

Technical aspects of creating an arteriovenous fistula in the mouse are discussed. Under general anesthesia, an abdominal incision is made, and the aorta ...

The Rabbit Blood-shunt Model for the Study of Acute and Late Sequelae of Subarachnoid Hemorrhage: Technical Aspects

Early brain injury and delayed cerebral vasospasm both contribute to unfavorable outcomes after subarachnoid hemorrhage (SAH). Reproducible and controllable animal models that simulate both conditions are presently uncommon. Therefore, new models are needed in order to mimic human pathophysiological conditions resulting from SAH.
This report describes the technical nuances of a rabbit blood-shunt SAH model that enables control of intracerebral pressure (ICP). An extracorporeal shunt is placed between the arterial system and the subarachnoid space, which enables examiner-independent SAH in a closed cranium. Step-by-step procedural instructions and necessary equipment are described, as well as technical considerations to produce the model with minimal mortality and morbidity. Important details required for successful surgical creation of this robust, simple and consistent ICP-controlled SAH rabbit model are described.

The experimental intracranial pressure-controlled blood shunt subarachnoid hemorrhage (SAH) model in the rabbit combines the standard procedures &#8212; subclavian artery cannulation and transcutaneous cisterna magna puncture, which enables close mimicking of human pathophysiological conditions after SAH. We present step-by-step instructions and discuss key surgical points for successful experimental SAH creation.

El Conejo Modelo Blood-shunt para el Estudio de la aguda y secuelas tardías de Hemorragia subaracnoidea: Aspectos Técnicos

Early brain injury and delayed cerebral vasospasm both contribute to unfavorable outcomes after subarachnoid hemorrhage (SAH). Reproducible and ...

A Simple Critical-sized Femoral Defect Model in Mice

While bone has a remarkable capacity for regeneration, serious bone trauma often results in damage that does not properly heal. In fact, one tenth of all limb bone fractures fail to heal completely due to the extent of the trauma, disease, or age of the patient. Our ability to improve bone regenerative strategies is critically dependent on the ability to mimic serious bone trauma in test animals, but the generation and stabilization of large bone lesions is technically challenging. In most cases, serious long bone trauma is mimicked experimentally by establishing a defect that will not naturally heal. This is achieved by complete removal of a bone segment that is larger than 1.5 times the diameter of the bone cross-section. The bone is then stabilized with a metal implant to maintain proper orientation of the fracture edges and allow for mobility. 
Due to their small size and the fragility of their long bones, establishment of such lesions in mice are beyond the capabilities of most research groups. As such, long bone defect models are confined to rats and larger animals. Nevertheless, mice afford significant research advantages in that they can be genetically modified and bred as immune-compromised strains that do not reject human cells and tissue.
Herein, we demonstrate a technique that facilitates the generation of a segmental defect in mouse femora using standard laboratory and veterinary equipment. With practice, fabrication of the fixation device and surgical implantation is feasible for the majority of trained veterinarians and animal research personnel. Using example data, we also provide methodologies for the quantitative analysis of bone healing for the model.

Animal models are frequently employed to mimic serious bone injury in biomedical research. Due to their small size, establishment of stabilized bone lesions in mice are beyond the capabilities of most research groups. Herein, we describe a simple method for establishing and analyzing experimental femoral defects in mice.

A-crítico tamaño femoral Defecto sencillo modelo en ratones

While bone has a remarkable capacity for regeneration, serious bone trauma often results in damage that does not properly heal. In fact, one tenth of all ...

Murine Model of Femoral Artery Wire Injury with Implantation of a Perivascular Drug Delivery Patch

Percutaneous interventions including balloon angioplasty and stenting have been used to restore blood flow in vessels with occlusive vascular disease. While these therapies lead to the rapid restoration of blood flow, these technologies remain limited by restenosis in the case of bare metal stents and angioplasty, or reduced healing and possibly enhanced risk of thrombosis in the case of drug eluting stents. A key pathophysiological mechanism in the formation of restenosis is intimal hyperplasia caused by the activation of vascular smooth muscle cells and inflammation due to arterial stretch and injury. Surgeries that induce arterial injury in genetically modified mice are useful for the mechanistic study of the vascular response to injury but are often technically challenging to perform in mouse models due to the their small size and lack of appropriate sized devices. We describe two approaches for a surgical technique that induces endothelial denudation and arterial stretch in the femoral artery of mice to produce robust neointimal hyperplasia. The first approach creates an arteriotomy in the muscular branch of the femoral artery to obtain vascular access. Following wire injury this arterial branch is ligated to close the arteriotomy. A second approach creates an arteriotomy in the main femoral artery that is later closed through localized cautery. This method allows for vascular access through a larger vessel and, consequently, provides a less technically demanding procedure that can be used in smaller mice. Following either method of arterial injury, a degradable drug delivery patch can be placed over or around the injured artery to deliver therapeutic agents.

We describe a surgical technique that produces wire injury in the femoral artery of mice to induce neointimal hyperplasia to serve as a model testing system for the perivascular delivery of therapeutic compounds for the inhibition of restenosis.

Modelo murino de la arteria femoral Lesiones Alambre con implantación de un perivascular Drug Delivery Patch

Percutaneous interventions including balloon angioplasty and stenting have been used to restore blood flow in vessels with occlusive vascular disease. ...

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, ...

Induction of Accelerated Atherosclerosis in Mice: The "Wire-Injury" Model

Atherosclerosis is a proliferative fibro-inflammatory disease developing in the arterial wall, inducing a deficient blood flow or a lack of blood flow. Moreover, by rupture of the defective vascular wall, atherosclerosis induces occlusive thrombus formation, which represents the main cause of myocardial infarction or stroke and the most frequent cause of death. Despite the advances in the cardiovascular field, many questions remain unanswered, and additional basic research is essential to improve our understanding of the molecular mechanisms during atherosclerosis and its effects. Due to limited clinical studies, there is a need for representative animal models recreating atherosclerotic conditions such as neointima formation after stent implantation, balloon angioplasty, or endarterectomy. Since the mouse presents many advantages and is the most frequently used model for studying molecular processes, the current study proposes an invasive procedure of endothelial denudation, also known as the wire-injury model, which is representative of the human condition of neointima formation in arteries after revascularization procedures.

Induction of Accelerated Atherosclerosis in Mice: The &quot;Wire-Injury&quot; Model

This study describes an invasive procedure for the induction of accelerated atherosclerosis in mice. In comparison to other methods using electric- or cryo-induced injury, mechanical-induced injury mimics the human condition of restenosis after revascularization therapies and is ideal for the study of the molecular mechanisms involved.

Inducción de la Aterosclerosis Acelerada en Ratones: El Modelo de "Lesiones por Alambre"

Atherosclerosis is a proliferative fibro-inflammatory disease developing in the arterial wall, inducing a deficient blood flow or a lack of blood flow. ...

Open Tracheostomy Gastric Acid Aspiration Murine Model of Acute Lung Injury Results in Maximal Acute Nonlethal Lung Injury

Acid pneumonitis is a major cause of sterile acute lung injury (ALI) in humans. Acid pneumonitis spans the clinical spectrum from asymptomatic to acute respiratory distress syndrome (ARDS), characterized by neutrophilic alveolitis, and injury to both alveolar epithelium and vascular endothelium. Clinically, ARDS is defined by acute onset of hypoxemia, bilateral patchy pulmonary infiltrates and non-cardiogenic pulmonary edema. Human studies have provided us with valuable information about the physiological and inflammatory changes in the lung caused by ARDS, which has led to various hypotheses about the underling mechanisms. Unfortunately, difficulties determining the etiology of ARDS, as well as a wide range of pathophysiology have resulted in a lack of critical information that could be useful in developing therapeutic strategies.
Translational animal models are valuable when their pathogenesis and pathophysiology accurately reproduce a concept proven in both in vitro and clinical settings. Although large animal models (e.g., sheep) share characteristics of the anatomy of human trachea-bronchial tree, murine models provide a host of other advantages including: low cost; short reproductive cycle lending itself to greater data acquisition; a well understood immunologic system; and a well characterized genome leading to the availability of a variety of gene deletion and transgenic strains. A robust model of low pH induced ARDS requires a murine ALI that targets mainly the alveolar epithelium, secondarily the vascular endothelium, as well as the small airways leading to the alveoli. Furthermore, a reproducible injury with wide differences between different injurious and non-injurious insults is important.
The murine gastric acid aspiration model presented here using hydrochloric acid employs an open tracheostomy and recreates a pathogenic scenario that reproduces the low pH pneumonitis injury in humans. Additionally, this model can be used to examine interaction of a low pH insult with other pulmonary injurious entities (e.g., food particles, pathogenic bacteria).

This protocol induces acute lung injury in a mouse that has close fidelity to the pathogenesis of acid pneumonitis observed in humans. We generate a maximal acute nonlethal low pH lung injury and account for differences in rodent-human anatomic respiratory structure using an open tracheostomy coupled with circumferential pressure release.

La traqueostomía abierta ácido gástrico aspiración modelo murino de lesión pulmonar aguda Resultados en máxima no letal aguda lesión pulmonar

Acid pneumonitis is a major cause of sterile acute lung injury (ALI) in humans. Acid pneumonitis spans the clinical spectrum from asymptomatic to acute ...