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.

Déclaration de l&#39;éthique: NOTE: Toutes les procédures sont conformes au Guide pour le soin et l&#39;utilisation des animaux de laboratoire (National Academies Press, 8e édition, 2011), et les protocoles approuvés par le Comité de protection des animaux et l&#39;utilisation institutionnelle à l&#39;Université du Wisconsin. Toutes les interventions chirurgicales ont été réalisées sous anesthésie à l&#39;isoflurane (par inhalation, débit 2 ml / min), et tous les efforts ont été faits pour minimiser la souffrance. Les animaux ont été euthanasiés dans une chambre progressivement rempli de CO 2. 1. L&#39;induction de l&#39;anesthésie générale et incision de la peau <ol><li> Utilisez mâle C57BL / 6 avec un âge de 12 à 16 semaines et le poids de 23-28 g. Anesthésier l&#39;animal par l&#39;isoflurane à travers un appareil d&#39;anesthésie en circuit fermé. Confirmez anesthésie appropriée en pinçant son orteil. Appliquer un peu de larmes pommade artificielle sur les yeux pour prévenir la sécheresse. </li><li> Placez la souris dans une position couchée avec le membre postérieur gauche légèrement enlevé et l&#39;articulation du genou slightly fléchie. Raser les cheveux autour de la partie supérieure de la cuisse gauche et désinfecter la peau avec de chlorhexidine, appliqué trois fois à l&#39;aide de coton stérile applicateurs à pointe. Utilisez un stérilisateur à billes chaude pour garder les instruments stérilisés pendant la chirurgie. </li><li> Placez un drap de gaze stérile sur la jambe gauche. Faire une incision de la peau longitudinale 1,5 cm droite sur la cuisse gauche médial de l&#39;articulation du genou à l&#39;aine (figure 1A). </li></ol> 2. L&#39;exposition de l&#39;artère fémorale et Branches REMARQUE: la figure 1B montre l&#39;anatomie du système vasculaire brute dans le haut de la cuisse. L&#39;artère fémorale superficielle (AFS) peut être observée à travers un microscope à dissection lorsque l&#39;incision de la peau est correctement établie le long de la ligne médiane de la cuisse médiale. <ol><li> Ouvrez l&#39;aponévrose mince recouvrant la SFA avec des pincettes fines et ensuite en boucle avec la SFA 9-0 suture en nylon. NOTE: Dans ce processus, le nerf fémoral est facilement mis enide, mais la veine fémorale adjacente nécessite une attention particulière parce que son mur est si mince qu&#39;il perfore facilement et saigne. La technique est essentiel de se concentrer sur l&#39;artère elle-même; tant que la dissection est réalisée tout près de l&#39;artère, la veine tout dommage accidentel est évitable. </li><li> Une fois la SFA est bouclée, maintenez la suture avec une pince de moustiques et tirez-le légèrement vers le bas. Cette manœuvre permet en outre la dissection plus facile, mais pour éviter d&#39;endommager l&#39;artère ne tirez pas trop fort. </li><li> Continuer dissection de l&#39;artère proximale pour exposer et boucle l&#39;artère fémorale commune (CFA) avec 9-0 suture en nylon. NOTE: Habituellement, bouclage de la CFA à un point en aval de l&#39;artère épigastrique (figure 1B) est assez pour l&#39;insertion de fil. Maintenez la boucle de suture CFA avec une pince de moustiques de la même façon. </li><li> Tirez la boucle de suture CFA vers le haut et la boucle de suture SFA vers le bas pour découvrir la bifurcation de SFA et l&#39;artère fémorale profonde (DFAE). Le DFAE, la branche principale de la CFA,va vers le muscle de la cuisse (figure 1B). NOTE: Le fil sera inséré de la DFA tard. </li><li> Boucle de la DFA avec prudence parce que la veine adjacente est emmêlé sous l&#39;artère. En cas de saignement de la veine, un 30 à 60 sec compression simple avec de la gaze est généralement suffisante pour produire l&#39;hémostase. </li><li> Une fois que le DFAE est bouclée avec 9-0 suture en nylon, assurez-vous tous les trois boucles de suture sont à leurs positions correctes (figure 2A). </li><li> Procéder à la dissection de la DFA distale pour ligaturer l&#39;artère en un point aussi loin que possible de la bifurcation. Couper suture supplémentaire, mais laisser une certaine longueur de sorte que les deux extrémités peuvent être tenus avec des pinces de moustiques, qui stabilisent le DFAE que l&#39;insertion de fil plus facile. </li></ol> 3. Création fil blessures <ol><li> Insérez fil via DFA <ol><li> Égoutter les sutures CFA, SFA et DFA d&#39;établir un hémostatique temporaire. </li><li> Faire un petit trou dans le DFA avec microciseaux. Insérez les conseils des courbes micropince pour dilater le trou assez grand pour un fil d&#39;un diamètre de 0,015 pouces (figure 2B montre la différence de taille entre l&#39;artère et le fil). <ol><li> Avant de faire un trou dans le DFA, desserrer la boucle de suture CFA et doucement serrer / presser le DFAE plusieurs fois avec micropince ternes pointe. Cette manœuvre élargit le diamètre artériel remarquable pour faciliter l&#39;insertion de fil. Humidifiant à fond le fil avec du PBS est également important pour une insertion en douceur. </li><li> Lorsque vous faites un trou dans le DFAE, ne pas recadrer la paroi artérielle, mais faire un petit pincement en forme de V et pincer le rabat avec micropince bout pointu pour la manipulation. </li></ol></li><li> Insérez le fil dans le trou suffisamment dilaté sur la DFA. Avancer doucement le fil vers l&#39;artère iliaque tout en desserrant la boucle de suture autour de la CFA. 5-8 mm est suffisant pour couvrir toute la longueur CFA dans la plupart des cas (figure 2C </strong&gt;). Ne pas insérer le fil de force comme une perforation artérielle se traduit généralement par la perte de la souris. Raccourcir le fil à environ 15 mm de longueur aide à la manutention. </li></ol></li><li> Laisser le fil à l&#39;intérieur pour une Min <ol><li> Après avoir inséré le fil à la longueur optimale, laisser pendant 1 min de surexploiter la paroi artérielle. Humidifiez le fil et les artères avec du PBS pour éviter le dessèchement. Un autre suture 9-0 en nylon peut être placé autour de la DFA proximale au point d&#39;insertion de fil pendant ce temps d&#39;attente. Cette suture sera utilisé plus tard pour fermer le trou sur le DFAE après le retrait du fil. </li></ol></li><li> Récupérer fil et reprendre le flux sanguin <ol><li> Maintenez le fil avec une pince navire de cathétérisme, et tirez doucement le dos; le fil doit être récupéré en douceur. Pour surmonter toute résistance, généralement causée par l&#39;adhésion du fil à la paroi artérielle, tirant le fil avec une légère rotation est utile. Encore une fois, une manœuvre de force pourrait entraîner la perte de l&#39;animal. </li><li> Unprès avoir récupérer le fil, ligaturer les DFA au point de bifurcation avec une suture 9-0 en nylon. Ne pas ligaturer trop près de la bifurcation, ce qui mettrait en péril l&#39;écoulement du sang à la SFA. </li><li> Desserrez les boucles de suture SFA et CFA de reprendre le flux sanguin normal. Notez que le CFA est bien élargi par insertion de fil (figure 2D). </li></ol></li></ol> 4. peau fermeture et Soins postopératoires <ol><li> Contrôle saigne. Suintement mineur peut être arrêté par compression simple; certains saignements majeurs artérielles nécessiteront points avec une suture de nylon 11-0. Ces artères suturées finiront par entraîner une occlusion artérielle. </li><li> Fermez la peau enroulée avec agrafeuses cutanées (habituellement 3-4 agrafes). Gardez la surface de la plaie stérilisé par application de la solution de chlorhexidine chirurgicale. </li><li> Appliquer les traitements post-opératoires. Contrôler la douleur de la souris avec des injections sous-cutanées de la buprénorphine. Garder la souris sur le coussin chauffant jusqu&#39;à ce qu&#39;il ait retrouvé sufFicient conscience de maintenir décubitus sternal. Maison dans une cage propre, sans compagnie d&#39;autres animaux jusqu&#39;à ce que complètement rétabli. </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.

La procédure de blessures de fil est applicable à toutes les souches de souris aussi longtemps que leurs structures anatomiques sont similaires. 4 Dans cet article, nous avons utilisé mâle C57BL / 6 avec un âge de 12 à 16 semaines. Comme nous l&#39;avons appris de nos études préliminaires, les artères fémorales de souris âgés de moins de 10 semaines sont souvent si petite que l&#39;insertion de fil est assez difficile. D&#39;autre part, l&#39;insertion du fil dans des souris âgées de 16 semaines...

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;">Fixed Core Wire Guide</td>
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			<td height="21" style="height:21px;width:216px;">Dilation Forceps</td>
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			<td>Curved, blunt tips</td>
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a murine model of arterial restenosis: technical aspects of femoral wire injury

La maladie cardiovasculaire causée par l&#39;athérosclérose est la principale cause de décès dans le monde développé. Rétrécissement de la lumière du vaisseau, en raison du développement de la plaque d&#39;athérome ou la rupture de plaques établies, interrompt le flux sanguin normal menant à diverses morbidités telles que l&#39;infarctus du myocarde et accident vasculaire cérébral. Dans les procédures endovasculaires cliniques telles que l&#39;angioplastie sont couramment effectuées à rouvrir la lumière. Cependant, ces traitements inévitablement endommager la paroi du récipient ainsi que l&#39;endothélium vasculaire, le déclenchement d&#39;une réponse de cicatrisation excessive et le développement d&#39;une plaque néo-intimale qui se étend dans la lumière provoquant la resténose des vaisseaux (re-rétrécissement). Resténose reste une cause majeure de l&#39;échec des traitements endovasculaires pour l&#39;athérosclérose. Ainsi, les modèles animaux précliniques de resténose sont d&#39;une importance vitale pour l&#39;étude des mécanismes physiopathologiques ainsi que la traduction des approches aux interventions vasculaires. Parmi plusieurs experime murinmodèles NTAL, blessures de fil de l&#39;artère fémorale est largement acceptée comme le plus approprié pour les études de la resténose post-angioplastie parce qu&#39;il ressemble étroitement à la procédure d&#39;angioplastie qui blesse deux endothélium et paroi de la cuve. Cependant, de nombreux chercheurs ont de la difficulté en utilisant ce modèle en raison de son haut degré de difficulté technique. Ce est principalement à cause d&#39;un fil métallique doit être insérée dans l&#39;artère fémorale, ce qui est environ trois fois plus mince que le fil, pour générer une blessure suffisamment importante pour induire la néo-intima. Ici, nous décrivons les détails chirurgicaux essentiels pour surmonter efficacement les difficultés techniques majeures de ce modèle. En suivant les procédures présentées, effectuer la blessure de fil de l&#39;artère fémorale de la souris devient plus facile. Une fois familiarisé, toute la procédure peut être terminée en 20 min.

Quatre semaines après la chirurgie, récoltent l&#39;artère fémorale fil blessés et font un spécimen de blocs de paraffine pour analyser la formation de néo-intima. Induire une anesthésie générale comme décrit ci-dessus. Ouvrir la poitrine immédiatement et insérer une aiguille de papillon de calibre 20 au ventricule gauche. Connectez la ligne à une bouteille de perfusion. Effectuer perfusion fixation par perfusion goutte à goutte de PBS suivi par 4% de paraformaldéhyde (généralement de 10 à 20...

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

La souris fil de l&#39;artère fémorale modèle de lésion de resténose est techniquement difficile. Dans ce protocole, nous détaillons les principaux détails techniques essentiels à l&#39;exécution avec succès des blessures de fil pour induire la néo-intima cohérente pour les études de la resténose.

Un modèle murin de la resténose artérielle: les aspects techniques de fémur fil blessures

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

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JoVE Journal

Medicine

19.9K vues.  University of Wisconsin School of Medicine and Public Health.  La souris fil de l'artère fémorale modèle de lésion de resténose est techniquement difficile. Dans ce protocole, nous détaillons les principaux détails techniques essentiels à l'exécution avec succès des blessures de fil pour induire la néo-intima cohérente pour les études de la resténose. 

Vidéo: 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.

Microchirurgie veineuse Pouch artériel Bifurcation anévrismes dans le modèle du lapin: Aspects techniques

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

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Médecine

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.

Fémorale cathétérisme artériel et veineux pour le prélèvement sanguin, Drug Administration et la pression artérielle Conscient et Mesures du rythme cardiaque

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 modèle murin d&#39;implantation d&#39;une endoprothèse dans l&#39;artère carotide pour l&#39;étude de la resténose

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.

Aspects techniques de la fistule aorto-cave de la souris

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.

Le lapin de sang-shunt modèle pour l&#39;étude de aiguë et les séquelles tardives de hémorragie méningée: Aspects techniques

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.

Une simple critique de taille fémorale défauts modèle chez la souris

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.

Modèle murin de l&#39;artère fémorale fil des blessures à l&#39;implantation d&#39;un périvasculaire 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.

Modèle murin de Intestinal Ischémie-reperfusion

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.

Induction de l’athérosclérose accélérée chez les souris : le modèle « Wire-Injury »

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.

Ouvrir trachéotomie acide gastrique Aspiration modèle murin de lésion pulmonaire aiguë Résultats dans Maximal aiguë non létaux 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 ...