Los autores agradecen a Christiane Pahrmann para su asistencia técnica.
D.W. fue apoyado por el Max Kade Foundation. T.D. recibidas becas de la Fundación de Kröner otro (2012_EKES.04) y la Deutsche Forschungsgemeinschaft (DE2133/2-1_. S. S. recibió becas de investigación de la Deutsche Forschungsgemeinschaft (DFG; SCHR992/3-1, SCHR992/4-1).

Animales recibieron atención humanitaria conforme a la guía de principios de los animales de laboratorio, prepararon por el Instituto de recursos de animales laboratorio y publicaron por los institutos nacionales de salud. Animales todos los protocolos fueron aprobados por la autoridad local responsable ('' Amt für Gesundheit und Verbraucherschutz, Hansestadt (Oficina de salud y protección del consumidor) Hamburgo '').
1. preparación de catéter
Nota: Consulte la Tabla de materiales para obtener información sobre el catéter.
<ol><li>Tomar el catéter del soporte del catéter.</li>
	<li>Tire de la guía de la luz a través del orificio distal hacia fuera.</li>
	<li>Poner en una gota de cianoacrilato en el extremo distal del catéter. PRECAUCIÓN: Use guantes de látex o nitrilo.</li>
	<li>Coloque la guía en el puerto de RX del catéter y avanzarlo a través del lumen hasta el puerto distal. Luego, tire la guía ligeramente para dejar un espacio de ~ 5 m m hasta el final.</li>
	<li>Espere 5 minutos para permitir que el adhesivo se seque.</li>
	<li>Mueva la guía, debe ser fijo. Si es aún móvil, repita los pasos 1.3-1.6.</li>
	<li>Llene una jeringa de 3 mL con solución salina al de 0.9% 1.5 mL y conéctelo en el puerto de inflado del globo.</li>
	<li>Empuje el émbolo de la jeringa para comprobar la inflación del globo. Salir de 0.6 mL solución salina en la jeringa.</li>
</ol>2. preparación de Mouse
<ol><li>Obtener ratones machos a la edad de 14 semanas aproximadamente 30 g de peso. Nota: Se utilizaron animales obtenidos desde el Instituto de animales de laboratorio. Aquí utilizamos el C57BL/6J.</li>
	<li>Utilizar una cámara de inducción para anestesiar el ratón con 2.0-2.5% isofurane (tasa de flujo de oxígeno de 500 mL/min).</li>
	<li>Coloque el ratón sobre su espalda en un cojín de calefacción y mantener la anestesia con una máscara que cubre la boca y la nariz del ratón. Si hay suficiente profundidad de la anestesia por pellizcar las patas traseras y la cola para verificar la ausencia de los reflejos.</li>
	<li>Eliminar el vello abdominal utilizando un cortador de pelo.</li>
	<li>Extiende las piernas traseras y fijar su posición con cinta.</li>
	<li>Desinfectar la zona abdominal con povidona-yodo, seguido de etanol al 80%. Repita este paso dos veces más.</li>
	<li>Utilice un paño quirúrgico para asegurarse no contaminado Haz sitio quirúrgico. Abrir las capas de piel y músculo a lo largo de la linea alba con una tijera (o bisturí) para exponer los órganos abdominales.</li>
	<li>Poner los intestinos en un guante hidratada solución salina 0.9% y envolver para mantenerlos húmedos.</li>
	<li>Use dos pinzas finas para quitar el tejido fino graso por encima de la aorta abdominal.</li>
	<li>Utilizando una jeringa de insulina (30G), inyectar 250 μl de solución de heparina (50 U/mL) en la Vena Cava Inferior (IVC) y espere 3 minutos para su distribución sistémica. Heparina se suprimen la hemostasia y prevenir la coagulación no deseada durante la cirugía.</li>
	<li>Utilizando dos pinzas, disecar la aorta del infrarenal hasta su bifurcación y sus vasos de rama saliente.</li>
	<li>Ligar los vasos laterales, que deben colocarse en la zona sujeta, con un cauterizador de alta temperatura.</li>
	<li>Detener el flujo de sangre por pinzamiento de la aorta del infrarenal directamente debajo de las arterias renales.</li>
	<li>Coloque la segunda abrazadera en una posición distal justo por encima de la bifurcación aórtica.</li>
	<li>Realizar una pequeña incisión horizontal con unas tijeras en el punto medio entre las abrazaderas a lo largo de la nave. Nota: El tamaño de la incisión debe ser equivalente a 1/3 de la circunferencia del vaso.</li>
	<li>Inserte una jeringa con una aguja (30G) en la incisión y al ras de la aorta con solución de heparina de 250 μl (50 U/mL).</li>
	<li>Utilizando suturas 10-0, coloque un nudo a cada lado de la incisión.</li>
	<li>Dilatación de la aorta por inserción de un dilatador de vasos en la incisión y extendió el vaso ligeramente. Repetir la dilatación 2 a 3 veces.</li>
	<li>Hidratar el catéter de balón con solución salina al 0.9%.</li>
	<li>Inserte el catéter de globo aplanado en la aorta y avanzar hacia la pinza proximal en la aorta.</li>
	<li>Al llegar a la pinza proximal, cuidadosamente abra la pinza proximal e inflar el balón para evitar fugas de sangre, inyectando ~0.6 mL de solución salina. Nota: La relación entre el globo inflado al recipiente es 1.5:1.</li>
	<li>Haga avanzar el catéter retrógrado de aproximadamente 2 cm.</li>
	<li>Tire el catéter ampliado, al desinflar el balón ligeramente por la liberación de la jeringa.</li>
	<li>Vuelva a colocar la pinza proximal cuando el catéter alcanza la incisión de la aorta. Desinflar el balón completamente y sáquela.</li>
	<li>Enjuague la aorta con solución de heparina de 250 μl (50 U/mL) usando una jeringa de 30G.</li>
	<li>Cerrar la incisión aórtica usando suturas 10-0. Lugar interrumpe puntadas en cada lado lateral, seguido por una o dos puntadas en el lado ventral.</li>
	<li>Abra la pinza distal. En caso de sangrado, vuelva a cerrar la pinza y colocar puntos de sutura adicionales.</li>
	<li>Abrir con cuidado la pinza proximal.</li>
	<li>Coloque dos hisopos en la sutura para apoyarlo y detener cualquier sangrado.</li>
	<li>Colocar pinzas hemostáticas absorbibles sobre la sutura para mantenerla. Nota: Un pulso aórtico debe ser visible distal de la incisión.</li>
	<li>Coloque los intestinos en el abdomen.</li>
	<li>Lavar la cavidad abdominal con solución salina 0.9% estéril, que ha sido precalentado a 37 ° C.</li>
	<li>Cerrar la capa de músculo abdominal usando suturas corrientes 6-0.</li>
	<li>Cierre la piel con suturas corrientes 5-0.</li>
	<li>Inyectar en forma subcutánea 4-5 mg/kg de carprofeno antes de permitir que el ratón despertar. Seguimiento del animal hasta que se ha ganado conciencia, mantener recumbency esternal. No el animal solo en una jaula hasta su recuperación completa.</li>
	<li>Añadir químicos al agua de bebida (50 mg/100 mL) como medicamento para el dolor por 3 días y monitorear diariamente el animal. Generalmente ratones como el dulce sabor de Metamizol y comenzar a beber inmediatamente después de la cirugía. Si lo prefiere, pueden utilizarse agentes inyectables de liberación sostenida en lugar de químicos. Nota: El período de observación para este modelo es de 28 días.</li>
</ol>3. histopatología
<ol><li>La cosecha la aorta lesionado globo después de 28 días preparando los ratones como se describe en los pasos 2.2 a 2.9.</li>
	<li>Utilice tijeras para quitar la aorta lesión de globo (entre la bifurcación y 0,3 mm por encima de los vasos renales) y eutanasia el ratón cortando a su corazón.</li>
	<li>Irrigue el lumen del vaso con 0.9% NaCl.</li>
	<li>Fijar la embarcación cosechada en paraformaldehído al 4% (PFA) durante la noche y deshidratar en el aumento de las concentraciones de etanol, a partir de etanol al 70% de etanol al 80% durante 1 hora, 2 horas, etanol al 95% durante 2 horas y el etanol al 100% durante 5 horas. Luego, incubar las muestras en xileno durante 2 horas 3 veces, antes de infiltrarse en las muestras con parafina. Nota: en lugar de rubor el buque cosechado con 0.9% NaCl, puede aclararse con 4% PFA. PRECAUCIÓN: PFA y el xileno son tóxicos y deben manipularse con especial cuidado.</li>
	<li>Embeber la muestra en parafina y cortadas en rodajas de 5 μm de espesor utilizando un micrótomo.</li>
	<li>Desparafinizar el portaobjetos con xileno 3 veces durante 5 minutos.</li>
	<li>Rehidratar las diapositivas de tejidos utilizando una serie decreciente de etanol. Iniciar con 100% etanol 2 veces durante 5 min, seguido de 3 minutos de etanol 95%, 80% y 70%.</li>
	<li>Mancha de los portaobjetos con tinción tricrómica de Masson como descrito18.</li>
	<li>Deshidratar diapositivas manchadas en etanol al 100% 2 veces por 10 minutos cada uno. Claro con xileno 2 veces de 10 minutos cada uno y Monte en medio de montaje.</li>
	<li>Ver las diapositivas con un microscopio de campo brillante. Utilizar un objetivo con 5 aumentos y una apertura numérica de 0.12 para un cuadro de resumen o una lente de 20 aumentos x con una apertura numérica de 0.35 para observación detallada.</li>
</ol>4. microscopia de la inmunofluorescencia de
<ol><li>Rehidratar las diapositivas de tejidos utilizando una serie decreciente de etanol. Iniciar con 100% etanol 2 veces durante 5 min, seguido de 3 minutos de etanol 95%, 80% y 70%.</li>
	<li>Realizar recuperación de antígeno por calentar los portaobjetos en solución de recuperación de antígeno en un vapor durante 20 minutos.</li>
	<li>Deje que las diapositivas se enfríe a temperatura ambiente.</li>
	<li>Después de lavar los portaobjetos tres veces con tampón fosfato salino (PBS), aplicar solución de bloqueo de antígeno en secciones de 30 minutos.</li>
	<li>Lave el portaobjetos tres veces durante 5 minutos con PBS.</li>
	<li>Incubar las secciones con anticuerpo primario diluido en un diluyente de anticuerpo primario. Nota: El momento de concentración y la incubación debe ser elegido por separado para cada anticuerpo.</li>
	<li>Lave el portaobjetos tres veces durante 5 minutos con PBS para retirar el anticuerpo no Unido.</li>
	<li>Incubar las secciones con un anticuerpo secundario conjugado previamente diluido en un diluyente de anticuerpo secundario. Nota: El momento de concentración y la incubación debe ser elegido por separado para cada anticuerpo.</li>
	<li>Eliminar los anticuerpos no Unidos por el lavado de los portaobjetos durante 5 minutos tres veces.</li>
	<li>Contratinción los núcleos celulares con 4', 6-diamidino-2-phenylindole (DAPI) durante 15 minutos; la concentración final de DAPI debe ser 350 nM.</li>
	<li>Montar diapositivas en inmunofluorescencia compatible con solución de montaje. Nota: Usando la solución de montaje incorrecto puede oscurecer la señal de fluorescencia.</li>
	<li>Diapositivas de la vista con un microscopio de fluorescencia. Use un lente de aumento de 40 x con una apertura numérica de 1.3.</li>
</ol>

<ol>
	<li>Kochanek, K. D., Xu, J., Murphy, S. L., Minino, A. M., Kung, H. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Deaths:+final+data+for+2009.">Deaths: final data for 2009.</a> Natl Vital Stat Rep. 60 (3), 1-116 (2011).</li><li>Austin, G. E., Ratliff, N. B., Hollman, J., Tabei, S., Phillips, D. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intimal+proliferation+of+smooth+muscle+cells+as+an+explanation+for+recurrent+coronary+artery+stenosis+after+percutaneous+transluminal+coronary+angioplasty.">Intimal proliferation of smooth muscle cells as an explanation for recurrent coronary artery stenosis after percutaneous transluminal coronary angioplasty.</a> J Am Coll Cardiol. 6 (2), 369-375 (1985).</li><li>Greenwald, S. E., Berry, C. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Improving+vascular+grafts:+the+importance+of+mechanical+and+haemodynamic+properties.">Improving vascular grafts: the importance of mechanical and haemodynamic properties.</a> J Pathol. 190 (3), 292-299 (2000).</li><li>Majesky, M. W., Schwartz, S. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Smooth+muscle+diversity+in+arterial+wound+repair.">Smooth muscle diversity in arterial wound repair.</a> Toxicol Pathol. 18 (4 Pt 1), 554-559 (1990).</li><li>Owens, G. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Regulation+of+differentiation+of+vascular+smooth+muscle+cells.">Regulation of differentiation of vascular smooth muscle cells.</a> Physiol Rev. 75 (3), 487-517 (1995).</li><li>Owens, G. K., Kumar, M. S., Wamhoff, B. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Molecular+regulation+of+vascular+smooth+muscle+cell+differentiation+in+development+and+disease.">Molecular regulation of vascular smooth muscle cell differentiation in development and disease.</a> Physiol Rev. 84 (3), 767-801 (2004).</li><li>Kleemann, R., Zadelaar, S., Kooistra, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cytokines+and+atherosclerosis:+a+comprehensive+review+of+studies+in+mice.">Cytokines and atherosclerosis: a comprehensive review of studies in mice.</a> Cardiovasc Res. 79 (3), 360-376 (2008).</li><li>Karas, S. P., 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=Coronary+intimal+proliferation+after+balloon+injury+and+stenting+in+swine:+an+animal+model+of+restenosis.">Coronary intimal proliferation after balloon injury and stenting in swine: an animal model of restenosis.</a> J Am Coll Cardiol. 20 (2), 467-474 (1992).</li><li>Ip, J. H., 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=The+role+of+platelets,+thrombin+and+hyperplasia+in+restenosis+after+coronary+angioplasty.">The role of platelets, thrombin and hyperplasia in restenosis after coronary angioplasty.</a> J Am Coll Cardiol. 17 (6 Suppl B), 77B-88B (1991).</li><li>Mason, R. G., Read, M. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Some+species+differences+in+fibrinolysis+and+blood+coagulation.">Some species differences in fibrinolysis and blood coagulation.</a> J Biomed Mater Res. 5 (1), 121-128 (1971).</li><li>Lafont, A., Faxon, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Why+do+animal+models+of+post-angioplasty+restenosis+sometimes+poorly+predict+the+outcome+of+clinical+trials?.">Why do animal models of post-angioplasty restenosis sometimes poorly predict the outcome of clinical trials?.</a> Cardiovasc Res. 39 (1), 50-59 (1998).</li><li>Matter, C. 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=Increased+balloon-induced+inflammation,+proliferation,+and+neointima+formation+in+apolipoprotein+E+(ApoE)+knockout+mice.">Increased balloon-induced inflammation, proliferation, and neointima formation in apolipoprotein E (ApoE) knockout mice.</a> Stroke. 37 (10), 2625-2632 (2006).</li><li>Lindner, V., Fingerle, J., Reidy, M. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mouse+model+of+arterial+injury.">Mouse model of arterial injury.</a> Circ Res. 73 (5), 792-796 (1993).</li><li>Simon, D. I., 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=Decreased+neointimal+formation+in+Mac-1(-/-)+mice+reveals+a+role+for+inflammation+in+vascular+repair+after+angioplasty.">Decreased neointimal formation in Mac-1(-/-) mice reveals a role for inflammation in vascular repair after angioplasty.</a> J Clin Invest. 105 (3), 293-300 (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>Moroi, 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=Interaction+of+genetic+deficiency+of+endothelial+nitric+oxide,+gender,+and+pregnancy+in+vascular+response+to+injury+in+mice.">Interaction of genetic deficiency of endothelial nitric oxide, gender, and pregnancy in vascular response to injury in mice.</a> J Clin Invest. 101 (6), 1225-1232 (1998).</li><li>Painter, T. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Myointimal+hyperplasia:+pathogenesis+and+implications.+2.+Animal+injury+models+and+mechanical+factors.">Myointimal hyperplasia: pathogenesis and implications. 2. Animal injury models and mechanical factors.</a> Artif Organs. 15 (2), 103-118 (1991).</li><li>Stubbendorff, 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=Inducing+myointimal+hyperplasia+versus+atherosclerosis+in+mice:+an+introduction+of+two+valid+models.">Inducing myointimal hyperplasia versus atherosclerosis in mice: an introduction of two valid models.</a> J Vis Exp. (87), e51459 (2014).</li><li>Deuse, T., 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=Dichloroacetate+prevents+restenosis+in+preclinical+animal+models+of+vessel+injury.">Dichloroacetate prevents restenosis in preclinical animal models of vessel injury.</a> Nature. 509 (7502), 641-644 (2014).</li></ol>

Los autores no tienen nada que revelar.

Este artículo demuestra un modelo murino para el desarrollo de la hiperplasia myointimal y permite la exploración de los procesos patológicos subyacentes y la prueba de nuevos medicamentos o terapéuticas.
El paso más crítico en este protocolo es la denudación de la aorta. Debe prestarse especial cuidado durante este paso como denudación excesiva dará lugar a la formación de aneurismas y el fracaso del modelo. Por otro lado, si la denudación se realiza escaso, desarrollará myointima...

Estenosis arterial en las arterias coronarias y periféricas tiene un gran efecto sobre la morbilidad y la mortalidad de pacientes1. Un mecanismo patológico subyacente es la hiperplasia myointima (MH), que se caracteriza por aumento de la proliferación, migración y síntesis de matriz extracelular proteínas del músculo liso vascular células (SMC)2. SMC se encuentran en la capa media del vaso y migrar sobre el estímulo a la superficie de la luz. Señales estimulantes incluyen factores de crecimiento, citoquinas, contacto célula-célula, lípidos, componentes de la matriz extracelular y esquileo mecánico y estiran las fuerzas3,4,5,6. Lesiones de la pared del vaso, patológica o iatrogénica, causan la célula endotelial y daño a las células del músculo liso y estimulan reacciones inflamatorias y así llevan a MH7.
Diferentes modelos animales están disponibles para el estudio de lesión arterial y la hiperplasia myointima. Grandes animales como cerdos o perros tienen la ventaja de compartir una similar de la arteria y la anatomía coronaria con los seres humanos y son especialmente adecuados para estudios investigando técnicas, procedimiento y dispositivos de angioplastia8. Sin embargo, modelos de cerdo tienen el inconveniente de mayor trombogenicidad9,10, mientras que los perros sólo tienen una suave respuesta a lesiones de vasos11. Además, todos los modelos animales grandes requieren especial, equipos y personal, que está relacionado con altos costos y no siempre está disponibles en una institución. Modelos animales pequeños incluyen ratas y ratones. En comparación con las ratas, ratones tienen las ventajas de menor costo y la existencia de una variedad de golpes modelos. El modelo descrito en este video se puede combinar con ApoE-/-los ratones alimentados con una dieta occidental que mímico de cerca el ajuste clínico de la angioplastia en los vasos ateroscleróticos12. Los modelos anteriores inducida por lesión vascular vía cable lesión13, desecación líquido14, primavera15o manguito lesiones16. Puesto que la naturaleza de la lesión afectará grandemente el desarrollo y Constitución de MH, utilizando un catéter con balón para inducir lesión de vaso es la mejor manera de imitar el ajuste clínico.
En este artículo describimos un nuevo método para inducir a MH con un catéter con balón en ratones. El uso de un catéter con balón (1,2 mm x 6 mm) con un puerto de RX (figura 1A) permite el raspado de la capa íntima y, al mismo tiempo, la inducción de un overdistension del buque. Ambos de estos factores son importantes factores desencadenantes para el desarrollo de MH. El tiempo de observación para este modelo es de 28 días17.

<table>
	<tbody>
		<tr>
			<td>10-0 Ethilon suture</td>
			<td>Ethicon</td>
			<td>2814G</td>
			<td></td>
		</tr>
		<tr>
			<td>3 mL Syringe</td>
			<td>BD Medical</td>
			<td>309658</td>
			<td></td>
		</tr>
		<tr>
			<td>37% HCl</td>
			<td>Sigma-Aldrich</td>
			<td>H1758</td>
			<td></td>
		</tr>
		<tr>
			<td>5-0 prolene suture</td>
			<td>Ethicon</td>
			<td>EH7229H</td>
			<td></td>
		</tr>
		<tr>
			<td>6-0 prolene suture</td>
			<td>Ethicon</td>
			<td>8706H</td>
			<td></td>
		</tr>
		<tr>
			<td>Acid Fuchsin</td>
			<td>Sigma-Aldrich</td>
			<td>F8129-25G</td>
			<td>Trichrome staining</td>
		</tr>
		<tr>
			<td>Antigen retrieval solution</td>
			<td>Dako</td>
			<td>S1699</td>
			<td></td>
		</tr>
		<tr>
			<td>Azophloxin</td>
			<td>Waldeck</td>
			<td>1B-103</td>
			<td>Trichrome staining</td>
		</tr>
		<tr>
			<td>Bepanthen Eye and Nose ointment</td>
			<td>Bayer</td>
			<td>1578675</td>
			<td>Eye ointment</td>
		</tr>
		<tr>
			<td>Betadine Solution</td>
			<td>Betadine Purdue Pharma</td>
			<td>NDC:67618-152</td>
			<td></td>
		</tr>
		<tr>
			<td>C57BL/6J</td>
			<td>Charles River</td>
			<td>Stock number 000664</td>
			<td></td>
		</tr>
		<tr>
			<td>Clamp applicator</td>
			<td>Fine Science Tools</td>
			<td>18056-14</td>
			<td></td>
		</tr>
		<tr>
			<td>Collagen 3</td>
			<td>abcam</td>
			<td>ab7778</td>
			<td>Antibody</td>
		</tr>
		<tr>
			<td>DAPI</td>
			<td>Thermo Fischer</td>
			<td>D1306</td>
			<td></td>
		</tr>
		<tr>
			<td>Donkey anti-Goat IgG AF555</td>
			<td>Invitrogen</td>
			<td>A21432</td>
			<td>Secondary antibody</td>
		</tr>
		<tr>
			<td>Donkey anti-Rabbit IgG AF488</td>
			<td>Invitrogen</td>
			<td>A21206</td>
			<td>Secondary antibody</td>
		</tr>
		<tr>
			<td>Donkey anti-Rabbit IgG AF488</td>
			<td>Invitrogen</td>
			<td>A11055</td>
			<td>Secondary antibody</td>
		</tr>
		<tr>
			<td>Donkey anti-Rabbit IgG AF555</td>
			<td>Invitrogen</td>
			<td>A31572</td>
			<td>Secondary antibody</td>
		</tr>
		<tr>
			<td>Ethanol 70%</td>
			<td>Th. Geyer</td>
			<td>2270</td>
			<td></td>
		</tr>
		<tr>
			<td>Ethanol 96%</td>
			<td>Th. Geyer</td>
			<td>2295</td>
			<td></td>
		</tr>
		<tr>
			<td>Ethanol absolute</td>
			<td>Th. Geyer</td>
			<td>2246</td>
			<td></td>
		</tr>
		<tr>
			<td>FAP</td>
			<td>abcam</td>
			<td>ab28246</td>
			<td>Antibody</td>
		</tr>
		<tr>
			<td>Forceps fine</td>
			<td>Fine Science Tools</td>
			<td>11251-20</td>
			<td></td>
		</tr>
		<tr>
			<td>Forceps standard</td>
			<td>Fine Science Tools</td>
			<td>11023-10</td>
			<td></td>
		</tr>
		<tr>
			<td>Glacial Acetic Acid</td>
			<td>Sigma-Aldrich</td>
			<td>537020</td>
			<td></td>
		</tr>
		<tr>
			<td>Hair clipper</td>
			<td>WAHL</td>
			<td>8786-451A ARCO SE</td>
			<td></td>
		</tr>
		<tr>
			<td>Heparin</td>
			<td>Rotexmedica</td>
			<td>PZN 3862340</td>
			<td>25.000 I.E./mL</td>
		</tr>
		<tr>
			<td>High temperature cautery kit</td>
			<td>Bovie</td>
			<td>18010-00</td>
			<td></td>
		</tr>
		<tr>
			<td>Image-iT FX Signal Enhancer</td>
			<td>Invitrogen</td>
			<td>I36933</td>
			<td>Blocking solution</td>
		</tr>
		<tr>
			<td>Light Green SF</td>
			<td>Waldeck</td>
			<td>1B-211</td>
			<td>Trichrome staining</td>
		</tr>
		<tr>
			<td>Microsurgical clamp</td>
			<td>Fine Science Tools</td>
			<td>18055-04</td>
			<td>Micro-Serrefine - 4mm</td>
		</tr>
		<tr>
			<td>MINI TREK Coronary Dilatation Catheter 1.20 mm x 6 mm / Rapid-Exchange</td>
			<td>Abbott</td>
			<td>1012268-06U</td>
			<td></td>
		</tr>
		<tr>
			<td>Molybdatophosphoric acid hydrate</td>
			<td>Merck</td>
			<td>1.00532.0100</td>
			<td>Trichrome staining</td>
		</tr>
		<tr>
			<td>NaCl 0,9%</td>
			<td>B.Braun</td>
			<td>PZN 06063042 Art. Nr.: 3570160</td>
			<td></td>
		</tr>
		<tr>
			<td>Needle holder</td>
			<td>Fine Science Tools</td>
			<td>12075-14</td>
			<td></td>
		</tr>
		<tr>
			<td>Novaminsulfon</td>
			<td>Ratiopharm</td>
			<td>PZN 03530402</td>
			<td>Metamizole</td>
		</tr>
		<tr>
			<td>Orange G</td>
			<td>Waldeck</td>
			<td>1B-221</td>
			<td>Trichrome staining</td>
		</tr>
		<tr>
			<td>Paraffin</td>
			<td>Leica biosystems</td>
			<td>REF 39602004</td>
			<td></td>
		</tr>
		<tr>
			<td>PBS pH 7,4</td>
			<td>Gibco</td>
			<td>10010023</td>
			<td></td>
		</tr>
		<tr>
			<td>PFA 4%</td>
			<td>Electron Microscopy Sciences</td>
			<td>#157135S</td>
			<td></td>
		</tr>
		<tr>
			<td>Ponceau S solution</td>
			<td>Serva Electrophoresis</td>
			<td>33427</td>
			<td>Trichrome staining</td>
		</tr>
		<tr>
			<td>Primary antibody diluent</td>
			<td>Dako</td>
			<td>S3022</td>
			<td></td>
		</tr>
		<tr>
			<td>Prolong Gold Mounting solution</td>
			<td>Thermo Fischer</td>
			<td>P36930</td>
			<td>Mounting solution for immunofluorescence stained slides</td>
		</tr>
		<tr>
			<td>Replaceable Fine Tip</td>
			<td>Bovie</td>
			<td>H101</td>
			<td></td>
		</tr>
		<tr>
			<td>Resorcin-Fuchsin Weigert</td>
			<td>Waldeck</td>
			<td>2E-30</td>
			<td>Trichrome staining</td>
		</tr>
		<tr>
			<td>Rimadyl</td>
			<td>Pfizer</td>
			<td>400684.00.00</td>
			<td>Carprofen</td>
		</tr>
		<tr>
			<td>Scissors</td>
			<td>Fine Science Tools</td>
			<td>14028-10</td>
			<td></td>
		</tr>
		<tr>
			<td>Scissors Vannas-style</td>
			<td>Fine Science Tools</td>
			<td>15000-03</td>
			<td></td>
		</tr>
		<tr>
			<td>Secondary antibody diluent</td>
			<td>Dako</td>
			<td>S0809</td>
			<td></td>
		</tr>
		<tr>
			<td>Fast acting Adhesive MINIS 3x1g</td>
			<td>UHU</td>
			<td>45370</td>
			<td>Cyanoacrylate</td>
		</tr>
		<tr>
			<td>Slide Rack</td>
			<td>Ted Pella</td>
			<td>21057</td>
			<td></td>
		</tr>
		<tr>
			<td>SM22</td>
			<td>abcam</td>
			<td>ab10135</td>
			<td>Antibody</td>
		</tr>
		<tr>
			<td>SMA</td>
			<td>abcam</td>
			<td>ab21027</td>
			<td>Antibody</td>
		</tr>
		<tr>
			<td>Staining dish</td>
			<td>Ted Pella</td>
			<td>21075</td>
			<td></td>
		</tr>
		<tr>
			<td>Surgical microscope</td>
			<td>Leica</td>
			<td>M651</td>
			<td></td>
		</tr>
		<tr>
			<td>Tabotamp fibrillar</td>
			<td>Ethicon</td>
			<td>431962</td>
			<td>Absorbable hemostat</td>
		</tr>
		<tr>
			<td>Transpore Surgical Tape</td>
			<td>3M</td>
			<td>1527-1</td>
			<td></td>
		</tr>
		<tr>
			<td>U-100 Insulin syringe</td>
			<td>BD Medical</td>
			<td>324825</td>
			<td></td>
		</tr>
		<tr>
			<td>Vessel Dilator</td>
			<td>Fine Science Tools</td>
			<td>18603-14</td>
			<td></td>
		</tr>
		<tr>
			<td>Vitro-Clud</td>
			<td>Langenbrinck</td>
			<td>04-0001</td>
			<td></td>
		</tr>
		<tr>
			<td>Weigerts iron hematoxylin Kit</td>
			<td>Merck</td>
			<td>1.15973.0002</td>
			<td>Trichrome staining</td>
		</tr>
		<tr>
			<td>Xylene</td>
			<td>Th. Geyer</td>
			<td>3410</td>
			<td></td>
		</tr>
	</tbody>
</table>

balloon-based injury to induce myointimal hyperplasia in the mouse abdominal aorta

El uso de modelos animales es esencial para una mejor comprensión de la MH, una de las principales causas de estenosis arterial. En este artículo, nos muestran un modelo de denudación murine del globo, que es comparable con modelos de lesión de buque establecido en grandes animales. El modelo de denudación de aorta con catéteres de balón imita el ajuste clínico y conduce a cambios fisiológicos y pathobiological comparables. Brevemente, después de realizar una incisión horizontal en la aorta abdominalis, un catéter con balón se introduce en el recipiente, inflado y se introdujo retrogradely. Inflación del balón dará lugar a lesiones de la íntima y overdistension del buque. Después de retirar el catéter, se cerrará la incisión aórtica con puntos de sutura únicas. El modelo mostrado en este artículo es reproducible, fácil de realizar y pueden establecerse de forma rápida y fiable. Es especialmente conveniente para la evaluación de agentes terapéuticos experimentales costosos, que pueden aplicarse de una manera económica. Mediante el uso de diferentes cepas de ratón knockout, podrá evaluarse el impacto de diferentes genes en el desarrollo de MH.

Denudación de globo es un modelo adecuado para estudiar el desarrollo de MH en ratones. Animales recuperan bien de la cirugía y mostrar después de la operación de una excelente condición física. Se estableció este modelo en 50 ratones con menos de 3% de mortalidad debido a la intervención quirúrgica. Figuras 1B -C muestra los principales pasos quirúrgicos. Después de una incisión en la piel a lo largo de la linea alba, id...

Watch this Scientific Journal Video about Balloon-based Injury to Induce Myointimal Hyperplasia in the Mouse Abdominal Aorta at JoVE.com

Balloon-based Injury to Induce Myointimal Hyperplasia in the Mouse Abdominal Aorta

Este artículo demuestra un modelo murino para el desarrollo de la hiperplasia myointimal (MH) después de lesión de globo.

Lesión basada en globo para inducir la hiperplasia Myointimal de la Aorta Abdominal de ratón

The use of animal models is essential for a better understanding of MH, one major cause for arterial stenosis.In this article, we demonstrate a murine ...

balloon-based-injury-to-induce-myointimal-hyperplasia-mouse-abdominal

Research

JoVE Journal

Medicine

9.5K vistas.  University Heart Center. Este artículo demuestra un modelo murino para el desarrollo de la hiperplasia myointimal (MH) después de lesión de globo.

Video: Balloon-based Injury to Induce Myointimal Hyperplasia in the Mouse Abdominal Aorta

The C-seal: A Biofragmentable Drain Protecting the Stapled Colorectal Anastomosis from Leakage

Colorectal anastomotic leakage (AL) is a serious complication in colorectal surgery leading to high morbidity and mortality rates1. The incidence of AL varies between 2.5 and 20% 2-5. Over the years, many strategies aimed at lowering the incidence of anastomotic leakage have been examined6, 7. 
The cause of AL is probably multifactorial. Etiological factors include insufficient arterial blood supply, tension on the anastomosis, hematoma and/or infection at the anastomotic site, and co-morbid factors of the patient as diabetes and atherosclerosis8. Furthermore, some anastomoses may be insufficient from the start due to technical failure.
Currently a new device is developed in our institute aimed at protecting the colorectal anastomosis and lowering the incidence of AL. This so called C-seal is a biofragmentable drain, which is stapled to the anastomosis with the circular stapler. It covers the luminal side of the colorectal anastomosis thereby preventing leakage.
The C-seal is a thin-walled tube-like drain, with an approximate diameter of 4 cm and an approximate length of 25 cm (figure 1). It is a tubular device composed of biodegradable polyurethane. Two flaps with adhesive tape are found at one end of the tube. These flaps are used to attach the C-seal to the anvil of the circular stapler, so that after the anastomosis is made the C-seal can be pulled through the anus. The C-seal remains in situ for at least 10 days. Thereafter it will lose strength and will degrade to be secreted from the body together with the gastrointestinal natural contents.
The C-seal does not prevent the formation of dehiscences. However, it prevents extravasation of faeces into the peritoneal cavity. This means that a gap at the anastomotic site does not lead to leakage.
Currently, a phase II study testing the C-seal in 35 patients undergoing (colo-)rectal resection with stapled anastomosis is recruiting. The C-seal can be used in both open procedures as well as laparoscopic procedures. The C-seal is only applied in stapled anastomoses within 15cm from the anal verge. In the video, application of the C-seal is shown in an open extended sigmoid resection in a patient suffering from diverticular disease with a stenotic colon.

The C-seal is a biofragmentable drain protecting the stapled colorectal anastomosis. In this video, details of the manufacturing, deployment and use are shown.

El C-sello: Un drenaje Biofragmentable protección de la anastomosis colorrectal grapada de fuga

Colorectal anastomotic leakage (AL) is a serious complication in colorectal surgery leading to high morbidity and mortality rates1. The incidence of AL ...

Investigación

Medicina

Pressure Controlled Ventilation to Induce Acute Lung Injury in Mice

Murine models are extensively used to investigate acute injuries of different organs systems (1-34). Acute lung injury (ALI), which occurs with prolonged mechanical ventilation, contributes to morbidity and mortality of critical illness, and studies on novel genetic or pharmacological targets are areas of intense investigation (1-3, 5, 8, 26, 30, 33-36). ALI is defined by the acute onset of the disease, which leads to non-cardiac pulmonary edema and subsequent impairment of pulmonary gas exchange (36). We have developed a murine model of ALI by using a pressure-controlled ventilation to induce ventilator-induced lung injury (2). For this purpose, C57BL/6 mice are anesthetized and a tracheotomy is performed followed by induction of ALI via mechanical ventilation. Mice are ventilated in a pressure-controlled setting with an inspiratory peak pressure of 45 mbar over 1 - 3 hours. As outcome parameters, pulmonary edema (wet-to-dry ratio), bronchoalveolar fluid albumin content, bronchoalveolar fluid and pulmonary tissue myeloperoxidase content and pulmonary gas exchange are assessed (2). Using this technique we could show that it sufficiently induces acute lung inflammation and can distinguish between different treatment groups or genotypes (1-3, 5). Therefore this technique may be helpful for researchers who pursue molecular mechanisms involved in ALI using a genetic approach in mice with gene-targeted deletion.

A murine model for ventilator induced lung injury is an important tool to study an acute lung injury in vivo. Here, we report an easy applicable in situ model for acute lung injury using high-pressure mechanical ventilation to induce acute failure of the lung.

La presión de la ventilación controlada para inducir la lesión pulmonar aguda en ratones

Murine models are extensively used to investigate acute injuries of different organs systems (1-34). Acute lung injury (ALI), which occurs with prolonged ...

The CYP2D6 Animal Model: How to Induce Autoimmune Hepatitis in Mice

Autoimmune hepatitis is a rare but life threatening autoimmune disease of the liver of unknown etiology1,2. In the past many attempts have been made to generate an animal model that reflects the characteristics of the human disease 3-5. However, in various models the induction of disease was rather complex and often hepatitis was only transient3-5. Therefore, we have developed a straightforward mouse model that uses the major human autoantigen in type 2 autoimmune hepatitis (AIH-2), namely hCYP2D6, as a trigger6. Type 1 liver-kidney microsomal antibodies (LKM-1) antibodies recognizing hCYP2D6 are the hallmark of AIH-27,8. Delivery of hCYP2D6 into wildtype FVB or C57BL/6 mice was by an Adenovirus construct (Ad-2D6) that ensures a direct delivery of the triggering antigen to the liver. Thus, the ensuing local inflammation generates a fertile field9 for the subsequent development of autoimmunity. A combination of intravenous and intraperitoneal injection of Ad-2D6 is the most effective route to induce a long-lasting autoimmune damage to the liver (section 1). Here we provide a detailed protocol on how autoimmune liver disease is induced in the CYP2D6 model and how the different aspects of liver damage can be assessed. First, the serum levels of markers indicating hepatocyte destruction, such as aminotransferases, as well as the titers of hCYP2D6 antibodies are determined by sampling blood retroorbitaly (section 2). Second, the hCYP2D6-specific T cell response is characterized by collecting lymphocytes from the spleen and the liver. In order to obtain pure liver lymphocytes, the livers are perfused by PBS via the portal vein (section 3), digested in collagen and purified over a Percoll gradient (section 4). The frequency of hCYP2D6-specific T cells is analyzed by stimulation with hCYP2D6 peptides and identification of IFN&gamma;-producing cells by flow cytometry (section 5). Third, cellular infiltration and fibrosis is determined by immunohistochemistry of liver sections (section 6). Such analysis regimen has to be conducted at several times after initiation of the disease in order to prove the chronic nature of the model. The magnitude of the immune response characterized by the frequency and activity of hCYP2D6-specific T and/or B cells and the degree of the liver damage and fibrosis have to be assessed for a subsequent evaluation of possible treatments to prevent, delay or abrogate the autodestructive process of the liver.

Infection of mice with an Adenovirus expressing the major human autoantigen cytochrome P450 2D6 (hCYP2D6) recognized by sera of patients suffering from type 2 autoimmune hepatitis results in a persistent form of autoimmune-mediated liver disease characterized by extensive hepatitis, fibrosis and generation of a CYP2D6-specific immune response.

El CYP2D6 Animal Modelo: cómo inducir a la hepatitis autoinmune en ratones

Autoimmune hepatitis is a rare but life threatening autoimmune disease of the liver of unknown etiology1,2. In the past many attempts have been made to ...

The Measurement and Treatment of Suppression in Amblyopia

Amblyopia, a developmental disorder of the visual cortex, is one of the leading causes of visual dysfunction in the working age population. Current estimates put the prevalence of amblyopia at approximately 1-3%1-3, the majority of cases being monocular2. Amblyopia is most frequently caused by ocular misalignment (strabismus), blur induced by unequal refractive error (anisometropia), and in some cases by form deprivation.
Although amblyopia is initially caused by abnormal visual input in infancy, once established, the visual deficit often remains when normal visual input has been restored using surgery and/or refractive correction. This is because amblyopia is the result of abnormal visual cortex development rather than a problem with the amblyopic eye itself4,5 . Amblyopia is characterized by both monocular and binocular deficits6,7 which include impaired visual acuity and poor or absent stereopsis respectively. The visual dysfunction in amblyopia is often associated with a strong suppression of the inputs from the amblyopic eye under binocular viewing conditions8. Recent work has indicated that suppression may play a central role in both the monocular and binocular deficits associated with amblyopia9,10 .
Current clinical tests for suppression tend to verify the presence or absence of suppression rather than giving a quantitative measurement of the degree of suppression. Here we describe a technique for measuring amblyopic suppression with a compact, portable device11,12 . The device consists of a laptop computer connected to a pair of virtual reality goggles. The novelty of the technique lies in the way we present visual stimuli to measure suppression. Stimuli are shown to the amblyopic eye at high contrast while the contrast of the stimuli shown to the non-amblyopic eye are varied. Patients perform a simple signal/noise task that allows for a precise measurement of the strength of excitatory binocular interactions. The contrast offset at which neither eye has a performance advantage is a measure of the "balance point" and is a direct measure of suppression. This technique has been validated psychophysically both in control13,14 and patient6,9,11 populations.
In addition to measuring suppression this technique also forms the basis of a novel form of treatment to decrease suppression over time and improve binocular and often monocular function in adult patients with amblyopia12,15,16 . This new treatment approach can be deployed either on the goggle system described above or on a specially modified iPod touch device15.

Amblyopia is a developmental disorder of the visual cortex that is often accompanied by strong suppression of one eye. We present a new technique for measuring and treating interocular suppression in patients with amblyopia that can be deployed using virtual reality goggles or a portable iPod Touch device.

La medición y tratamiento de la represión en la ambliopía

Amblyopia, a developmental disorder of the visual cortex, is one of the leading causes of visual dysfunction in the working age population. Current ...

An Alkali-burn Injury Model of Corneal Neovascularization in the Mouse

Under normal conditions, the cornea is avascular, and this transparency is essential for maintaining good visual acuity. Neovascularization (NV) of the cornea, which can be caused by trauma, keratoplasty or infectious disease, breaks down the so called &lsquo;angiogenic privilege' of the cornea and forms the basis of multiple visual pathologies that may even lead to blindness. Although there are several treatment options available, the fundamental medical need presented by corneal neovascular pathologies remains unmet. In order to develop safe, effective, and targeted therapies, a reliable model of corneal NV and pharmacological intervention is required. Here, we describe an alkali-burn injury corneal neovascularization model in the mouse. This protocol provides a method for the application of a controlled alkali-burn injury to the cornea, administration of a pharmacological compound of interest, and visualization of the result. This method could prove instrumental for studying the mechanisms and opportunities for intervention in corneal NV and other neovascular disorders.

Neovascularization (NV) of the cornea can complicate multiple visual pathologies. Utilizing a controlled, alkali-burn injury model, a quantifiable level of corneal NV can be produced for mechanistic study of corneal NV and evaluation of potential therapies for neovascular disorders.

Una Lesión álcalis quemar modelo de neovascularización corneal en el ratón

Under normal conditions, the cornea is avascular, and this transparency is essential for maintaining good visual acuity. Neovascularization (NV) of the ...

Inducing Myointimal Hyperplasia Versus Atherosclerosis in Mice: An Introduction of Two Valid Models

Various in vivo laboratory rodent models for the induction of artery stenosis have been established to mimic diseases that include arterial plaque formation and stenosis, as observed for example in ischemic heart disease. Two highly reproducible mouse models&#160;&#8211;&#160;both resulting in artery stenosis but each underlying a different pathway of development&#160;&#8211; are introduced here. The models represent the two most common causes of artery stenosis; namely one mouse model for each myointimal hyperplasia, and atherosclerosis are shown. To induce myointimal hyperplasia, a balloon catheter injury of the abdominal aorta is performed. For the development of atherosclerotic plaque, the ApoE -/- mouse model in combination with western fatty diet is used. Different model-adapted options for the measurement and evaluation of the results are named and described in this manuscript. The introduction and comparison of these two models provides information for scientists to choose the appropriate artery stenosis model in accordance to the scientific question asked.

This video shows two models of intimal plaque development in murine arteries and emphasizes the differences in myointimal hyperplasia and atherosclerosis.

Inducir miointimal Hiperplasia Versus aterosclerosis en ratones: una introducción de dos modelos válidos

Various in vivo laboratory rodent models for the induction of artery stenosis have been established to mimic diseases that include arterial plaque ...

Ischemic Tissue Injury in the Dorsal Skinfold Chamber of the Mouse: A Skin Flap Model to Investigate Acute Persistent Ischemia

Despite profound expertise and advanced surgical techniques, ischemia-induced complications ranging from wound breakdown to extensive tissue necrosis are still occurring, particularly in reconstructive flap surgery. Multiple experimental flap models have been developed to analyze underlying causes and mechanisms and to investigate treatment strategies to prevent ischemic complications. The limiting factor of most models is the lacking possibility to directly and repetitively visualize microvascular architecture and hemodynamics. The goal of the protocol was to present a well-established mouse model affiliating these before mentioned lacking elements. Harder et al. have developed a model of a musculocutaneous flap with a random perfusion pattern that undergoes acute persistent ischemia and results in ~50% necrosis after 10 days if kept untreated. With the aid of intravital epi-fluorescence microscopy, this chamber model allows repetitive visualization of morphology and hemodynamics in different regions of interest over time. Associated processes such as apoptosis, inflammation, microvascular leakage and angiogenesis can be investigated and correlated to immunohistochemical and molecular protein assays. To date, the model has proven feasibility and reproducibility in several published experimental studies investigating the effect of pre-, peri- and postconditioning of ischemically challenged tissue.

The window of the murine dorsal skinfold chamber presented visualizes a zone of acute persistent ischemia of a musculocutaneous flap. Intravital epi-fluorescence microscopy permits for direct and repetitive assessment of the microvasculature and quantification of hemodynamics. Morphologic and hemodynamic results can further be correlated with histological and molecular analyses.

Lesiones tejido isquémico en la Cámara de los pliegues cutáneos dorsal del ratón: Un modelo de la piel de la aleta para Investigar la isquemia aguda persistente

Despite profound expertise and advanced surgical techniques, ischemia-induced complications ranging from wound breakdown to extensive tissue necrosis are ...

Creation of Abdominal Adhesions in Mice

Abdominal adhesions consist of fibrotic tissue that forms in the peritoneal space in response to an inflammatory insult, typically surgery or intraabdominal infection. The precise mechanisms underlying adhesion formation are poorly understood. Many compounds and physical barriers have been tested for their ability to prevent adhesions after surgery with varying levels of success. The mouse and rat are important models for the study of abdominal adhesions. Several different techniques for the creation of adhesions in the mouse and rat exist in the literature. Here we describe a protocol utilizing abrasion of the cecum with sandpaper and sutures placed in the right abdominal sidewall. The mouse is anesthetized and the abdomen is prepped. A midline laparotomy is created and the cecum is identified. Sandpaper is used to gently abrade the surface of the cecum. Next, several figure-of-eight sutures are placed into the peritoneum of the right abdominal sidewall. The abdominal cavity is irrigated, a small amount of starch is applied, and the incision is closed. We have found that this technique produces the most consistent adhesions with the lowest mortality rate.

Abdominal adhesions that form after surgery are a major cause of pain, infertility, and hospitalization and reoperation for small bowel obstruction. Our surgical procedure for creating abdominal adhesions in mice is a reliable tool to study the mechanisms underlying the formation of adhesions.

Creación de abdominales adherencias en ratones

Abdominal adhesions consist of fibrotic tissue that forms in the peritoneal space in response to an inflammatory insult, typically surgery or ...

Reproducible Arterial Denudation Injury by Infrarenal Abdominal Aortic Clamping in a Murine Model

Percutaneous vascular interventions uniformly result in arterial denudation injuries that subsequently lead to thrombosis and restenosis. These complications can be attributed to impairments in re-endothelialization within the wound margins. Yet, the cellular and molecular mechanisms of re-endothelialization remain to be defined. While several animal models to study re-endothelialization after arterial denudation are available, few are performed in the mouse because of surgical limitations. This undermines the opportunity to exploit transgenic mouse lines and investigate the contribution of specific genes to the process of re-endothelialization. Here, we present a step-by-step protocol for creating a highly reproducible murine model of arterial denudation injury in the infrarenal abdominal aorta using external vascular clamping. Immunocytochemical staining of injured aortas for fibrinogen and &#946;-catenin demonstrate the exposure of a pro-thrombotic surface and the border of intact endothelium, respectively. The method presented here has the advantages of speed, excellent overall survival rate, and relative technical ease, creating a uniquely practical tool for imposing arterial denudation injury in transgenic mouse models. Using this method, investigators may elucidate the mechanisms of re-endothelialization under normal or pathological conditions.

Understanding the cellular and molecular mechanisms of re-endothelialization following arterial denudation injury is of paramount importance in preventing thrombosis and restenosis of arteries. Here we describe a protocol for reproducible arterial denudation injury of the infrarenal abdominal aorta. The procedure was developed to investigate the underlying mechanisms that regulate endothelial regeneration using mouse models.

Reproducible lesión arterial Denudación por pinzamiento aórtico abdominal infrarrenal en un modelo murino

Percutaneous vascular interventions uniformly result in arterial denudation injuries that subsequently lead to thrombosis and restenosis. These ...

A Model of Cardiac Remodeling Through Constriction of the Abdominal Aorta in Rats

Heart failure is one of the leading causes of death worldwide. It is a complex clinical syndromethat includes fatigue, dyspnea, exercise intolerance, and fluid retention. Changes in myocardial structure, electrical conduction, and energy metabolism develop with heart failure, leading to contractile dysfunction, increased risk of arrhythmias, and sudden death. Hypertensive heart disease is one of the key contributing factors of cardiac remodeling associated with heart failure. The most commonly-used animal model mimicking hypertensive heart disease is created via surgical interventions, such as by narrowing the aorta. Abdominal aortic constriction is a useful experimental technique to induce a pressure overload, which leads to heart failure. The surgery can be easily performed, without the need for chest opening or mechanical ventilation. Abdominal aortic constriction-induced cardiac pathology progresses gradually, making this model relevant to clinical hypertensive heart failure. Cardiac injury and remodeling can be observed 10 weeks after the surgery. The method described here provides a simple and effective approach to produce a hypertensive heart disease animal model that is suitable for studying disease mechanisms and for testing novel therapeutics.

A rat model of abdominal aortic constriction that induces cardiac hypertrophy and remodeling is described. An efficient, highly-reproducible, and minimally-invasive method is used to provide a simple yet useful platform for research in myocardial hypertrophy and dysfunction.

Un modelo de remodelamiento cardiaco A través de la constricción de la aorta abdominal en ratas

Heart failure is one of the leading causes of death worldwide. It is a complex clinical syndromethat includes fatigue, dyspnea, exercise intolerance, and ...