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.

HINWEIS: Ethics Statement: Alle Verfahren entsprechen dem Leitfaden für die Pflege und Verwendung von Labortieren (National Academies Press, 8. Auflage, 2011), und Protokolle von der Institutional Animal Care und Verwenden Committee an der University of Wisconsin zugelassen. Alle Operationen wurden unter Isofluran-Narkose (durch Einatmen, Durchflussrate von 2 ml / min) durchgeführt, und alle Anstrengungen unternommen, um das Leiden zu minimieren. Die Tiere wurden in einer Kammer allmählich mit CO 2 gefüllt euthanasiert. 1. Narkoseeinleitung und Hautschnitt <ol><li> Verwenden männlichen C57BL / 6-Mäuse mit einem Alter von 12-16 Wochen und das Gewicht 23-28 g. Anesthetize das Tier durch Isofluran über einen geschlossenen Kreisnarkosegerät. Überprüfen Sie die Betäubung durch Kneifen seine Zehen. Tragen Sie etwas künstliche Tränen Salbe auf die Augen bis zur Trockenheit zu verhindern. </li><li> Platzieren Sie die Maus in Rückenlage mit dem linken Hinterbein leicht verschleppt und Kniegelenk slightly gebeugt. Rasieren Sie die Haare um den linken Oberschenkel Region und desinfizieren Sie die Haut mit Chlorhexidin, angewendet dreimal mit sterilen Wattestäbchen Applikatoren. Verwenden Sie eine heiße Perle Sterilisator, die Instrumente während der Operation sterilisiert zu halten. </li><li> Legen Sie eine sterilisierte Gaze Tuch über das linke Bein. Einen 1,5 cm gerader Längs Hautinzision links medialen Oberschenkel vom Kniegelenk auf der Leiste (1A). </li></ol> 2. Exposition der Femoralarterie und Zweige HINWEIS: 1B zeigt die Bruttogefäßanatomie in den Oberschenkel. Die oberflächliche Oberschenkelarterie (SFA) können durch ein Präpariermikroskop beobachtet, wenn die Inzision richtig entlang der Mittellinie des medialen Oberschenkel vorgenommen werden. <ol><li> Öffnen Sie die dünne Faszie über dem SFA mit feinen Pinzette und dann die Schleife die SFA mit 9-0 Nylonnaht. HINWEIS: Bei diesem Verfahren wird der N. femoralis ist einfach wie setzenide, aber die angrenzenden femoralis erfordert besondere Aufmerksamkeit, weil ihre Wand ist so dünn, dass es leicht perforiert und blutet. Der Schlüssel Technik ist es, auf die Arterie selbst zu konzentrieren; solange der Präparation wird in der Nähe der Arterie durchgeführt wird, ist jede versehentliche Venenschäden vermeidbar. </li><li> Sobald die SFA geschlungen, halten Sie die Naht mit Moskito Pinzette und ziehen Sie sie leicht nach unten. Dieses Manöver macht weitere Präparation leichter, aber um eine Beschädigung der Arterie nicht zu kräftig ziehen. </li><li> Weiter sezieren die Arterie proximal zu entlarven und Schleife die A. femoralis communis (CFA) mit 9-0 Nylonnaht. HINWEIS: Normalerweise Schleifen des CFA an einem Punkt distal der A. epigastrica (1B) ist ausreichend für Drahteinführung. Halten Sie die CFA Fadenschleife mit Moskito Pinzette auf die gleiche Weise. </li><li> Ziehen Sie den CFA Fadenschleife nach oben und der SFA Fadenschleife nach unten, um die Gabelung der SFA und tiefe Oberschenkelarterie (EDA) zu entdecken. Das EDA, der Hauptzweig der CFA,fließt in den Oberschenkelmuskel (1B). HINWEIS: Der Draht wird später von der DFA eingefügt werden. </li><li> Schleife das EDA mit besonderer Vorsicht, da die angrenzenden Vene unter der Arterie verheddert. Im Falle von Blutungen aus der Vene ist ein 30- bis 60 sec einfache Kompression mit Gaze Regel ausreichend, um die Hämostase zu erzeugen. </li><li> Sobald der DFA mit 9-0 Nylonnaht geschleift, sicherzustellen, dass alle drei Nahtschleifen sind an die richtige Position (Abbildung 2A). </li><li> Fahren Sie mit der Dissektion der DFA distal, um die Arterie an einer Stelle so weit wie möglich von der Gabelung zu ligieren. Geschnitten zusätzliche Naht lassen aber eine bestimmte Länge, so dass die beiden Enden mit Moskito Pinzette, die die Herstellung von DFA Drahteinsetzlöcher leichter Stabilisierung statt. </li></ol> 3. Erstellen von Draht Injury <ol><li> Legen Draht über DFA <ol><li> Belastung der CFA, SFA und DFA Nähten, um eine temporäre hemostat etablieren. </li><li> Machen Sie ein kleines Loch in der DFEin mit Mikroschere. Stecken Sie die Spitzen der gebogenen Mikropinzetten, um das Loch groß genug für einen Draht mit einem 0,015 Zoll Durchmesser erweitern (2B zeigt den Größenunterschied zwischen der Arterie und dem Draht). <ol><li> Bevor Sie ein Loch in der DFA, lösen Sie die CFA Fadenschleife und sanft klemmen / Drücken Sie die DFA mehrmals mit stumpfen bestückte Mikropinzetten. Dieses Manöver erweitert die arteriellen Durchmesser bemerkenswert, um die Drahteinführung zu erleichtern. Befeuchten Sie den Draht sorgfältig mit PBS ist auch wichtig für eine reibungslose Einsetzen. </li><li> Wenn ein Loch in der DFA, nicht beschneiden die Arterienwand, aber einen kleinen V-förmigen Spalt und nehmen Sie die Klappe mit spitzen Mikropinzetten für Manipulation. </li></ol></li><li> Führen Sie den Draht durch die angemessen-dilatative Loch auf der DFA. Den Draht vorsichtig voran in Richtung der Darmbeinarterie, während das Lösen der Fadenschleife um das CFA. 5-8 mm reicht aus, um alle CFA Länge in den meisten Fällen zu decken (2C </strong&gt;). Haben Sie das Kabel mit Gewalt einlegen nicht als ein arterieller Perforation führt in der Regel zum Verlust der Maus. Die Verkürzung der Draht bis ca. 15 mm in der Länge hilft bei der Handhabung. </li></ol></li><li> Lassen Draht innen für 1 Min <ol><li> Nach dem Einlegen der Kabel an die optimale Länge, lassen Sie es für 1 Minute, um die Arterienwand überfordern. Befeuchten Sie den Draht und Arterien mit PBS, um ein Austrocknen zu verhindern. Eine weitere 9-0 Nylonnaht können rund um die DFA proximal der Drahteinführungspunkt während dieser Wartezeit platziert werden. Dieser Faden wird später verwendet werden, um das Loch auf der DFA nach Entfernen des Drahtes zu schließen. </li></ol></li><li> Abrufen Draht und Fortsetzen der Durchblutung <ol><li> Halten Sie den Draht mit Schiff Kanülierung Pinzetten, und ziehen Sie sie vorsichtig zurück; der Draht sollte problemlos abgerufen werden. Keinen Widerstand, in der Regel durch die Einhaltung des Drahtes an der Arterienwand, zu überwinden, das Zurückziehen des Drahtes mit leichter Rotation hilfreich. Auch hier könnte eine kraftvolle Manöver in der Tierverlust führen. </li><li> Aach das Abrufen der Draht, ligieren die DFA am Verzweigungspunkt mit 9-0 Nylonnaht. Ligieren Sie nicht zu nahe an der Gabelung, die den Blutabfluss an die SFA in Frage stellen würde. </li><li> Lösen Sie die SFA und CFA Nahtschleifen, um den normalen Blutfluss wieder aufzunehmen. Beachten Sie, dass die CFA wird durch Drahteinführung (2D) gut ausgebaut. </li></ol></li></ol> 4. Hautverschluss und postoperative Pflege <ol><li> Steuer blutet. Minor Nässen durch einfache Kompression gestoppt werden; einige große arterielle Blutungen wird Stiche mit einem 11-0 Nylonfaden erfordern. Diese vernäht Arterien schließlich in Arterienverschluss führen. </li><li> Schließen Sie die Haut mit Hautklammergeräte (in der Regel 3-4 Heftklammern) gewickelt. Halten Sie die Wundfläche durch Anwendung Chlorhexidin chirurgische Lösung sterilisiert. </li><li> Bewerben postoperative Behandlungen. Steuern Sie den Schmerz der Maus mit subkutanen Injektionen von Buprenorphin. Halten Sie die Maus auf die Erwärmung pad bis sie suf wiederreichend Bewusstsein Brustlage zu halten. Haus in einem sauberen Käfig ohne Begleitung anderer Tiere, bis sie vollständig erholt. </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.

Der Draht Verletzungsverfahren ist für alle Stämme von Mäusen, solange ihre anatomischen Strukturen sind ähnlich. 4 In diesem Dokument verwendet man männliche C57BL / 6-Mäuse mit einem Alter von 12-16 Wochen. Wie wir von unseren Vorstudien gelernt, sind die Oberschenkelarterien von Mäusen, die jünger als 10 Wochen oft so klein, dass Drahteinführung ist eine ziemliche Herausforderung. Andererseits ist Drahteinsetzlöcher in Mäusen, die älter als 16 Wochen technisch einfacher, aber neigt dazu, in var...

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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a murine model of arterial restenosis: technical aspects of femoral wire injury

Kardiovaskuläre Erkrankungen durch Arteriosklerose ist die häufigste Todesursache in der entwickelten Welt. Einengung des Gefäßlumens, aufgrund atherosklerotischen Plaque Entwicklung oder dem Aufbrechen der etablierten Plaques, unterbricht den normalen Blutfluss führt zu verschiedenen Begleiterkrankungen wie Herzinfarkt und Schlaganfall. In der Klinik endovaskulären Verfahren wie Angioplastie werden üblicherweise durchgeführt, um den Hohlraum wieder zu öffnen. Jedoch haben diese Behandlungen unvermeidbar beschädigt die Gefßwand sowie die Gefäßendothel Auslösen eines übermäßigen Heilungsreaktion und die Entwicklung einer neointimalen Plaque, die in das Lumen verursacht Gefäß Restenose (Wiederverengung) erstreckt. Restenose bleibt eine Hauptursache für Versagen endovaskulären Behandlungen für Atherosklerose. Somit sind vorklinischen Tiermodellen von Restenose für die Untersuchung der pathophysiologischen Mechanismen sowie Translationsansätze Gefäßinterventionen von entscheidender Bedeutung. Unter mehreren murinen Experimental Modellen wird Femoralarterie Drahtverletzung gilt als die am besten geeignete für die Untersuchung der Restenose nach Angioplastie akzeptiert, weil es ähnelt der Angioplastie-Verfahren, die sowohl Endothel und Gefäßwand verletzt. , Haben viele Forscher jedoch Schwierigkeiten Verwendung dieses Modell aufgrund seiner hohen technischen Schwierigkeitsgrad. Dies ist vor allem, weil ein Metalldraht muss in die Oberschenkelarterie, die etwa dreimal geringer als die Draht eingeführt werden, um eine ausreichende Verletzungen erzeugen prominente Neointima induzieren. Hier beschreiben wir die wesentlichen chirurgischen Details, die wichtigsten technischen Schwierigkeiten dieses Modells effektiv zu überwinden. Indem Sie die vorgestellten Verfahren, die Durchführung der Maus Femoralarterie Drahtverletzung wird einfacher. Sobald vertraut, kann das gesamte Verfahren innerhalb von 20 Minuten abgeschlossen werden.

Vier Wochen nach der Operation, zu ernten die Draht verletzten Oberschenkelarterie und machen einen Paraffinblock Probe, um die Neointimabildung analysieren. Induzieren Narkose wie oben beschrieben. Sofort Öffnen Sie die Kiste und legen Sie eine 20-Gauge-Butterfly-Nadel in die linke Herzkammer. Schließen Sie die Leitung an einer Infusionsflasche. Führen Perfusionsfixierung über Tropfinfusion von PBS, gefolgt von 4% Paraformaldehyd (in der Regel 10 bis 20 ml, jeweils). Die Infusionsflasche wird ca. 140 cm h?...

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

Die Maus Femoralarterie Drahtverletzungsmodell der Restenose ist technisch anspruchsvoll. In diesem Protokoll zeigen wir die wichtigsten technischen Details unerlässlich für die erfolgreiche Durchführung von Draht Verletzung konsequente Neointima für Studien der Restenose zu induzieren.

Ein Mausmodell von arterieller Restenose: Technische Aspekte der Femoral Draht Injury

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

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

Research

JoVE Journal

Medicine

19.9K Aufrufe.  University of Wisconsin School of Medicine and Public Health.  Die Maus Femoralarterie Drahtverletzungsmodell der Restenose ist technisch anspruchsvoll. In diesem Protokoll zeigen wir die wichtigsten technischen Details unerlässlich für die erfolgreiche Durchführung von Draht Verletzung konsequente Neointima für Studien der Restenose zu induzieren. 

Serie: 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.

Mikrochirurgische venöse Pouch Arterial-Bifurcation Aneurysmen in the Rabbit Modell: Technische Aspekte

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

Forschung

Medizin

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.

Femoral arterielle und venöse Katheterisierung zur Blutentnahme, Drug Administration und Conscious Blutdruck und Herzfrequenz-Messungen

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.

Ein Mausmodell der Stentimplantation in der Arteria carotis für das Studium der Restenose

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.

Technische Aspekte der Maus aortokavaler Fistel

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.

Das Kaninchen Blut-Shunt-Modell für die Untersuchung von akuten und Spätfolgen Subarachnoidalblutung: Technische Aspekte

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.

Eine einfache Kritische große Femoral Defect Modell an der Maus

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.

Mausmodell der Femoralarterie Draht Verletzung mit Implantation einer Perivaskuläre 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.

Murine Modell der intestinalen Ischämie-Reperfusionsschaden

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.

Induktion von beschleunigter Atherosklerose bei Mäusen: Das "Wire-Injury"-Modell

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.

Offene Tracheotomie Magensäure Aspiration Mausmodell der akuten Lungenschädigung Ergebnisse in Maximal Akute 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 ...