Diese Arbeit wurde von der Schweizer National Science Foundation Grant 150271 unterstützt.

dieses experimentelle Protokoll genehmigt wurde, durch das kantonale Veterinäramt, Freiburg und der Swiss Federal Veterinary Office, Schweiz (FR 2015/58).
 Hinweis: männliche NZW Kaninchen mit einem Gewicht von 2,8 bis 3,2 kg dienten. Die Tiere lebten unter herkömmlichen Bedingungen (12 h hell und dunkel-Zyklus, ad libitum Wasser und Essen zur Verfügung gestellt). Vor dem Ballon Denudation wurden Tiere für 1 Woche gewöhnt, in denen sie mit normalen Chow Diät gefüttert wurden. Nach 1 Woche der Akklimatisation Kaninchen auf atherogenen Diät bestehend aus fettreichen (8,6 %) umgestellt wurden, und gesättigte Fettsäuren mit 205 mg/kg (1 %) Cholesterin Diät für die gesamte Studiendauer. Ballon-Verletzung in der linken Beckenkamm Arterie war 1 Woche nach Beginn der Diät durchgeführt und nach 2 Wochen oder 4 Wochen Verletzungsgefahr Ballon Tiere geopfert wurden.
 1. präoperative Verfahren <ol><li>alle chirurgischen Instrumente vor Gebrauch mit einem Glas Bead Sterilisator oder andere geeignete Instrumente sterilisieren.</li>
	<li>Vorbereiten und überprüfen Sie die Ballon-Katheter-Montage.
	<ol><li>Anfügen einer 1 mL-Luer-Lock-Spritze gefüllt mit normalen Kochsalzlösung auf den Luer-Lock-Teil des Ballonkatheters. Abwesenheit der eingeschlossenen Luft überwachen Sie sorgfältig. Überprüfen Sie die Dichtheit und richtigen Ballon Inflation durch drücken den Kolben der Spritze zu gewährleisten.</li>
	</ol></li> <li>Wiegen die Kaninchen und schalten Sie die Thermopad auf 37 ° c</li>
	<li>Verwenden eine Buprenorphin-Lösung mit einer Konzentration von 0,3 mg / ml injizieren eine Dosis von 0,01 mg/kg subkutan.</li>
	<li>Anesthetize das Kaninchen mit 5 % Isofluran und 5 L/min O 2 in eine Induktion Kammer für 10-15 min.</li>
	<li>Legen die anästhesierten Kaninchen auf das Heizkissen auf die chirurgische Plattform gehalten. Legen Sie den Patch und Clips zur Überwachung der Temperatur, Atmung und Elektrokardiogramm.</li>
	<li>Legen die Schnauze des Kaninchens auf eine Gesichtsmaske an eine geeignete Anästhesie-Maschine angeschlossen. Aufrechterhaltung der Narkose mit Isofluran (4,0 % mit 2,5 L/min O 2). Richtige Anesthetization (gekennzeichnet durch Mangel an Muskeltonus und Verlust von Knebel und Ohrmuschel Reflexe) zu bestätigen.</li>
	<li>Gelten ophthalmologischen Salbe für beide Augen auf der Hornhaut vor dem Austrocknen zu verhindern. Drapieren Sie die Kaninchen mit einem sterilen chirurgischen Blatt mit nur der unteren Extremität ausgesetzt.</li>
</ol> 2. Chirurgische Protokoll <ol><li>Entfernen Sie die Haare aus dem ventralen Bereich direkt unterhalb der Kniegelenke mit tierischen Haarschneidemaschinen.</li>
	<li>Wischen Sie die Fläche mit einem geeigneten Desinfektionsmittel zu reinigen die Haut und entfernen lose Haare.</li>
	<li>Der saphenous Ader zu finden und machen einen kleinen Hautschnitt von ca. 1,5 cm Länge mit einem Skalpell.</li>
	<li>Setzen Sie einen kleinen Teil des saphenous Ader mit kleinen gebogenen Pinzette ohne Beschädigung der femoral Ader und femoral Nerv.</li>
	<li>Legen Sie zwei Lose Ligatur Schleifen (5-0 Seide) unterhalb der saphenous Ader und binden Sie eine Schleife der Ligatur am distalen Ende der Arterie. Legen Sie eine mikrovaskuläre Klammer über die Ligatur zu stoppen den Blutfluss aus dem Beckenkamm Arterie.</li>
	<li>Topisch angewendet einen Tropfen des Papaverine, die Arterie zu dehnen und zu verhindern Vasospasmus.</li>
	<li>Heben Sie die saphena Arterie mit Hilfe von gebundenen Ligatur und machen einen kleinen Arteriotomie Schnitt mit einem 24 gauge Nadel.</li>
	<li>Erhöhen die Schnitt-Klappen mit feinen Zangen und langsam Vene Abholung oder eine leitende Nadel in das Lumen der Arterie einfügen.</li>
	<li>Legen Sie 2 Französisch Fogarty arteriellen Embolectomy Katheter in der saphenous Ader. Entfernen Sie die Vene Pick- and -mikrovaskuläre Klemmen.</li>
	<li>Fördern den Katheter bis die sechste Mark (20-25 cm), eine Position entspricht etwa 2 bis 5 cm über dem Beckenkamm Bifurkation.</li>
	<li>Den Ballon aufblasen, mit 0,1 mL normale Kochsalzlösung mit einer 1 mL Spritze oder mit einem nominalen Druck von 6 atm mit einer geregelten manuelle Inflator, wie beschrieben in 16 , 22.</li>
	<li>Halten den Ballonkatheter mit Pinzette und ziehen Sie wieder um 6 cm über dem Beckenkamm Arterytoward Einfügepunkt beim Drehen des Katheters.</li>
	<li>Entleeren des Ballons durch den Kolben der Spritze zurückziehen.</li>
	<li>Wiederholen Sie die Schritte 2.10 bis 2.13 dreimal komplett endotheliale Denudation sicherzustellen.</li>
	<li>Den Katheter entfernen und sofort binden die Ligatur Schleife oberhalb der Arteriotomie Website um Blutungen zu stoppen.</li>
	<li>Gelten geeignet antiseptische alles rund um die Peripherie der Wunde und Tupfer entfernt das Blut gerinnt. Schließen Sie den Hautschnitt mit einem 5: 0-Naht, und desinfizieren der Chirurgie-Website mit Povidon-Jod-Lösung.</li>
	<li>Wiederholen Sie die Schritte 2.1 zu 2.16 auf der kontralateralen Beckenkamm mit einem neuen Katheter.</li>
	<li>Tupfer die ophthalmologische Salbe aus Augen.</li>
</ol> 3. Postoperative Versorgung <ol><li>verwalten Sulfadoxine 40 mg/kg und Trimethoprim 8 mg/kg oder jedes andere geeignete Antibiotikum unmittelbar nach dem chirurgischen Eingriff.</li>
	<li>Während der Anästhesie Erholungsphase, halten Sie die Kaninchen über ein Wärmekissen in einem sauberen autoklaviert Käfig platziert.</li>
	<li>Entfernen Sie die Überwachung Patch und Clips.</li>
	<li>Nach seiner Genesung die Kaninchen zu ihrer Heimat Käfige zurückkehren. Injektion subkutan Buprenorphin 0,05 - 0,1 mg/kg post - operativ alle 6-12 h für 48 h weiter atherogenen Diät für weitere zwei Wochen oder vier Wochen.</li>
</ol> 4. Gewebe-Ernte und Analyse von Plaque Zusammensetzung <ol><li>nach zwei Wochen (für frühe dünne Plaque) oder drei Wochen Verletzungsgefahr Ballon, die Kaninchen mit Isofluran in ähnlicher Weise wie oben beschrieben zu betäuben.</li>
	<li>Der Brusthöhle zu öffnen und die Kaninchen durch Intracardial Entbluten einschläfern.</li>
	<li>Isolieren die Beckenkamm Arterien, wie in 23 beschrieben.
	<ol><li>Kurz, öffnen Sie den Bauch und das Retroperitoneum setzen. Spur der Aorta gegenüber der Beckenkamm Bifurkation und binden Sie es oberhalb der Bifurkation. Entfernen Sie vorsichtig das umliegende Gewebe zu entlarven und isolieren beide Beckenkamm Arterien.</li>
	</ol></li> <li>Dissect aus beiden Beckenkamm Arterien und tauche sie in eiskalten Phosphat gepufferte Kochsalzlösung. Die Klumpen mit Hilfe der Zange zu entfernen. Teilen Sie jede Beckenkamm Arterie in 4-6 Segmente, die Dicke der Beläge in der Arterie zu charakterisieren.</li>
	<li>Sofort Einbetten der arteriellen Segmente in einer Form mit optimale Arbeitstemperatur zusammengesetzte, Snap-Freeze mit flüssigem Stickstoff und halten Sie es bei-70 ° C. bereiten 5 µm dicke Abschnitte mit einem Kryostaten, wie beschrieben in 24.</li>
	<li>Perform Histologie, Immunfluoreszenz oder immunhistochemischen Färbung für Morphometrie, Plaque Lipid und zelluläre Inhalt wie unter 10 , 25. 
	Hinweis: Kurz, spülen Sie die arteriellen Abschnitte mit Phosphat gepufferte Kochsalzlösung (PBS) und permeabilize mit 0,2 % Triton. Spülen Sie die Abschnitte mit PBS und blockieren Sie nicht bestimmte Websites mit 2 % Rinderserumalbumin für 30 min Inkubation in den Abschnitten für 1 h bei 37 ° C mit anti-α-SM Aktin (1: 200) oder RAM11 Antikörper (1: 200). Spülen Sie die Abschnitte mit PBS und inkubieren sie mit entsprechenden Sekundärantikörper für 30 min bei 37 ° c wieder mit PBS waschen und Hinzufügen von Hoechst (5 µg/mL) für 10 min, Kerne zu erkennen.</li>
</ol>

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A., Mosse, P. R., Safstrom, J. A., Young, S. B., Campbell, G. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Balloon+catheter+injury+to+rabbit+carotid+artery.+I.+Changes+in+smooth+muscle+phenotype.">Balloon catheter injury to rabbit carotid artery. I. Changes in smooth muscle phenotype.</a> Arteriosclerosis. 9, 289-298 (1989).</li><li>Miyake, 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=Prevention+of+neointimal+formation+after+angioplasty+using+nuclear+factor-kappaB+decoy+oligodeoxynucleotide-coated+balloon+catheter+in+rabbit+model.">Prevention of neointimal formation after angioplasty using nuclear factor-kappaB decoy oligodeoxynucleotide-coated balloon catheter in rabbit model.</a> Circ Cardiovasc Interv. 7, 787-796 (2014).</li><li>Fulcher, J., Patel, S., Nicholls, S. J., Bao, S., Celermajer, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Optical+coherence+tomography+for+serial+in+vivo+imaging+of+aortic+plaque+in+the+rabbit:+a+preliminary+experience.">Optical coherence tomography for serial in vivo imaging of aortic plaque in the rabbit: a preliminary experience.</a> Open Heart. 2, e000314 (2015).</li><li>Abela, O. G., 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=Plaque+Rupture+and+Thrombosis:+the+Value+of+the+Atherosclerotic+Rabbit+Model+in+Defining+the+Mechanism.">Plaque Rupture and Thrombosis: the Value of the Atherosclerotic Rabbit Model in Defining the Mechanism.</a> Curr Atheroscler Rep. 18, 29 (2016).</li><li>Yamashita, A., Asada, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+rabbit+model+of+thrombosis+on+atherosclerotic+lesions.">A rabbit model of thrombosis on atherosclerotic lesions.</a> J Biomed Biotechnol. 2011, 424929 (2011).</li></ol>

Die Autoren erklären keine konkurrierenden finanziellen Interessen.

Das Kaninchen Beckenkamm Arterie Arteriosklerose Modell ist in der Arteriosklerose Forschung verbreitet. Mit diesem Protokoll entwickelt die Kaninchen schnell schwere und erweiterte Plaques im Vergleich zu spontanen Läsionen mit Cholesterin-Diät entwickelt. Vor allem erholen Tiere schnell von der Operation.
Die wichtigsten Impulse für die Atherogenese ist die mechanische Schäden durch den Ballonkatheter, der verletzt des Endothels und glättet das Schiff Wand26. Die...

Bruch der gefährdeten arteriosklerotische Plaques gehört zu den häufigsten Todesursachen in den Industrienationen1. Obwohl Forschung in den letzten Jahrzehnten mehrere molekulare und zelluläre Mechanismen beteiligt Plaque Fortschreiten entfaltet hat, weiterhin Anstrengungen sind noch erforderlich, nicht nur die komplexen Mechanismen der Progression der Erkrankung zu entwirren, sondern auch neue testen therapeutische Ansätze. Verschiedenen Tiermodellen sind vorgeschlagen worden, um der Arteriosklerose zu studieren. Genmanipulation, Cholesterin Fütterung oder mechanische Endothel Verletzungen sind die standard Strategien von am meisten Tiermodelle der Atherosklerose einschließlich Mäuse, Kaninchen oder Minischweine geteilt. Unter diesen sind NZW Kaninchen empfindlich auf Cholesterin-Diät, während normale Ratten und Mäuse nicht signifikant Cholesterin2,3,4aufnehmen. Kaninchen entwickeln spontan Aorta Verletzungen reichen in Makrophagen mit einigen faserigen Komponente bei Fütterung mit Cholesterin-reiche Ernährung-5,-6. Die lange Vorlaufzeit von 4-8 Monaten induzieren atherosklerotischen Plaquesby Fütterung Cholesterin Diät allein6,7 ist jedoch einen großen Nachteil für die meisten experimentellen Einstellungen. In der Verfolgung zur Induktion Läsionen in relativ kurzer Zeit wurde eine Kombination aus hohen Cholesterin Diät und Ballon Verletzungen durch Baumgarter und Studer8entwickelt. Das übergeordnete Ziel dieser Technik ist, arteriosklerotische Plaques bestehend aus Schaumzellen (ähnlich wie fetthaltige Streifen beim Menschen) bei normalem Kaninchen innerhalb von 2 Wochen zu induzieren. Die gegenwärtige Technik beschreibt das Verfahren der Arterienwand Verletzung basierend auf Baumgarter Methode mit einem Ballonkatheter rückte in den Beckenkamm Arterie von normalem Kaninchen NZW.
Zusammen mit einem Cholesterin-reiche Diät werden Verletzungen durch induzierte Ballon de Endothelialization zu Arteriosklerose führen. Ballon-Verletzungen beschleunigt die Bildung von atherosklerotischen Läsionen und produziert Plaque von einheitlicher Größe und Verteilung. Intima-Verdickung steigt über einen Zeitraum von Zeit und Intima Zelle eindringen beginnt innerhalb weniger Tage nach Verletzung. Fetthaltige Streifen mit erheblichen Makrophagen beginnen, nach 7-10 Tagen Ballon Verletzungsgefahr zu erscheinen und sind als Typ-II-Läsion nach der Klassifikation von American Heart Association vertreten. Ballon-Verletzung in Kaninchen erfolgt häufig in der Aorta Plaque Komposition zu studieren. Das Endothel Neointimal drückt hohe interzelluläre Adhäsionsmolekül. Die Plaketten sind mediale Dissektion und adventitial Änderungen zugeordnet. Atherosklerotische Läsionen bestehen aus Lipiden, wuchernden glatten Muskelzellen (SMCs), Kollagenfasern und Entzündungszellen, die unter das regenerierte Endothel ansammeln und sind meist Typ II in der Natur. Die topologische Verteilung von Kaninchen Plaques war ähnlich wie in menschlichen Hauptschlagadern 9,10 im Prinzip berichtet, die Aorta ist größer im Vergleich zum Beckenkamm Arterien und Plaque in der größeren Länge produzieren würden. Der große Vorteil der Verwendung der Beckenkamm Arterie als Ort der Arteriosklerose bei Kaninchen ist jedoch ihre Zugänglichkeit, seine Ähnlichkeit im muskulären Inhalt menschliche Koronararterie11, einheitliche Läsion Entwicklung12, hohe Gewebe Faktor Aktivität13 und konsequente Schiff Dimension menschlichen Koronararterien ermöglicht die Bewertung der kommerziell hergestellten Geräte, morphometrische und angiographischen Endpunkte vergleichbar. Invasive und nicht-invasive Methoden wurden untersucht, um die Plaques in Kaninchen Beckenkamm Arterien im lebenden Tier zu analysieren. Frühere Berichte beschreiben die Verwendung von Magnet-Resonanz-Tomographie (MRT) mit Hilfe eines 2,35-Tesla Herr System 14 zusätzlich, intravaskulären Ultraschall (IVUS) oder optische Kohärenz Tomographie (OCT) Katheter können entsprechend auf Bild arteriosklerotische Plaques in Kaninchen Beckenkamm Arterien. Die Beckenkamm Arterie ist zugänglich für Ultraschall-Bildgebung bei Verwendung einer hochauflösenden Sonografie und die Aorta kann auch mit dieser Technik erkundet werden.
In den letzten zehn Jahren hat diese Kaninchen-Modell der Ballon Verletzungen um weiter zu verstehen, die Mechanismen der Plaque Fortschreiten15und Plaque Regression16. Darüber hinaus wurde das Modell verwendet, um die Untersuchung des Einflusses von neuartiger Therapeutika wie Statine, standard gerinnungshemmende Mittel, antioxidative Wirkstoffe17,18 und Medikamente freisetzende Stents wie Everolimus oder Zotarolimus-eluting Stent19,20 auf Neointimal Verdickung. Dieses Modell hat auch zur intravaskulären Bildgebung der Nah-Infrarot-Fluoreszenz imaging Katheter21zu untersuchen.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
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		<tr>
			<td>New Zealand White rabbits</td>
			<td>Charles River laboratories,France</td>
			<td>Cre:KBL(NZW)</td>
			<td></td>
		</tr>
		<tr>
			<td>Cholesterol rich diet</td>
			<td>Ssniff spezialdi&#228;ten</td>
			<td>Ssniff EF K High Fat and Cholesterol</td>
			<td></td>
		</tr>
		<tr>
			<td>Glass bead sterilizer-Germinator 500</td>
			<td>VWR, Leicestershire, UK</td>
			<td>101326-488</td>
			<td></td>
		</tr>
		<tr>
			<td>Fogarty balloon embolectomy catheters, 2 French</td>
			<td>Edwards Lifesciences, Switzerland</td>
			<td>120602F</td>
			<td>For single use only</td>
		</tr>
		<tr>
			<td>Luer Lock Syringe</td>
			<td>Becton, Dickinson and Company, USA</td>
			<td>309628</td>
			<td></td>
		</tr>
		<tr>
			<td>Thermopad Type 226</td>
			<td>Solis, Switzerland AG</td>
			<td>397387</td>
			<td></td>
		</tr>
		<tr>
			<td>Buprenorphine- Temgesic</td>
			<td>Reckitt Benckiser AG, Switzerland</td>
			<td>7.68042E+12</td>
			<td></td>
		</tr>
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			<td>Isoflurane</td>
			<td>Piramal Critical Care, Inc, Bethlehem, PA 18017</td>
			<td>2667-46-7</td>
			<td></td>
		</tr>
		<tr>
			<td>Anaesthesia machine-combi-vet Base Anesthesia System</td>
			<td>Rothacher Medical GmbH, Switzerland</td>
			<td>CV 30-301-A</td>
			<td></td>
		</tr>
		<tr>
			<td>Cardell touch veterinary vital signs monitor</td>
			<td>Midmark, Ohio, USA</td>
			<td>8013-001</td>
			<td></td>
		</tr>
		<tr>
			<td>Ophthalmic ointment-Humigel</td>
			<td>Virbac, France</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Animal hair clippers</td>
			<td>Aesculap AG, Germany</td>
			<td>GT420</td>
			<td></td>
		</tr>
		<tr>
			<td>Disinfectant-Betadine solution</td>
			<td>MundipharmaMedicalCompany, Switzerland</td>
			<td>14671-1203</td>
			<td></td>
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		<tr>
			<td>Dumont #7 Forceps</td>
			<td>FST Germany</td>
			<td>11274-20</td>
			<td></td>
		</tr>
		<tr>
			<td>Medium and small microscissors</td>
			<td>Medline International Switzerland S&#224;rl</td>
			<td>UC4337</td>
			<td></td>
		</tr>
		<tr>
			<td>Microvascular clamps</td>
			<td>FST, Germany</td>
			<td>18051-28</td>
			<td></td>
		</tr>
		<tr>
			<td>Papaverine</td>
			<td>ESCA chemicals, Switzerland</td>
			<td>RE 356 803</td>
			<td></td>
		</tr>
		<tr>
			<td>Vein Pick</td>
			<td>Harvard Apparatus, Cambridge, UK</td>
			<td>72-4169</td>
			<td>For single use only</td>
		</tr>
		<tr>
			<td>Saline</td>
			<td>Laboratorium Dr. G. Bichsel AG, , Switzerland</td>
			<td>1330055</td>
			<td></td>
		</tr>
		<tr>
			<td>Polysorb 5-0 suture</td>
			<td>Covidien AG, Switzerland</td>
			<td>UL 202</td>
			<td>Monofilament</td>
		</tr>
		<tr>
			<td>Sulfadoxine and Trimethoprim-Trimethazol</td>
			<td>Werner Stricker AG, Switzerland</td>
			<td>Swissmedic Nr. 50&#39;361</td>
			<td></td>
		</tr>
		<tr>
			<td>Antiseptic- Octenisept</td>
			<td>Sch&#252;lke &#38; Mayr AG, Switzerland</td>
			<td>GTIN: 4032651214068</td>
			<td></td>
		</tr>
		<tr>
			<td>Phosphate Buffered Saline</td>
			<td>Roth</td>
			<td>1058.1</td>
			<td></td>
		</tr>
		<tr>
			<td>Isobutanol-2-Methylbutane</td>
			<td>Sigma-Aldrich, Switzerland</td>
			<td>M32631-1L</td>
			<td></td>
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		<tr>
			<td>Optimum Cutting Temperature compound-Tissue-Tek</td>
			<td>VWR Chemicals, Belgium</td>
			<td>25608-930</td>
			<td></td>
		</tr>
		<tr>
			<td>Cryostat</td>
			<td>Leica, Glattbrugg, Switzerland</td>
			<td>Leica CM1860 UV</td>
			<td></td>
		</tr>
		<tr>
			<td>Glass slide- Superfrost Plus</td>
			<td>Thermo Scientific</td>
			<td>4951PLUS4</td>
			<td></td>
		</tr>
		<tr>
			<td>Mayer&#39;s Haematoxylin</td>
			<td>Sigma-Aldrich, Switzerland</td>
			<td>MHS32-1L</td>
			<td></td>
		</tr>
		<tr>
			<td>Eosin 0.5% aq.</td>
			<td>Sigma-Aldrich, Switzerland</td>
			<td>HT110232-1L</td>
			<td></td>
		</tr>
		<tr>
			<td>Oil Red O</td>
			<td>Sigma-Aldrich, Switzerland</td>
			<td>O0625-25G</td>
			<td></td>
		</tr>
		<tr>
			<td>&#945;-smooth muscle actin antibody</td>
			<td>Abcam, UK.</td>
			<td>ab7817</td>
			<td></td>
		</tr>
		<tr>
			<td>Macrophage Clone RAM11 antibody</td>
			<td>DAKO, Switzerland</td>
			<td>M063301</td>
			<td></td>
		</tr>
		<tr>
			<td>Hoechst</td>
			<td>Abcam, UK.</td>
			<td>ab145596</td>
			<td></td>
		</tr>
		<tr>
			<td>Goat polyclonal Secondary Antibody (Chromeo 546)</td>
			<td>Abcam, UK.</td>
			<td>ab60316</td>
			<td></td>
		</tr>
		<tr>
			<td>Alexa Fluor 488/547</td>
			<td>Abcam, UK.</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Glycergel Mounting Medium, Aqueous</td>
			<td>DAKO, Switzerland</td>
			<td>C056330</td>
			<td></td>
		</tr>
		<tr>
			<td>Hematoxylin for Movat pentachrome staining</td>
			<td>Sigma-Aldrich, Switzerland</td>
			<td>H3136-25G</td>
			<td></td>
		</tr>
		<tr>
			<td>Ferric chloride for Movat pentachrome staining</td>
			<td>Sigma-Aldrich, Switzerland</td>
			<td>157740-100G</td>
			<td></td>
		</tr>
		<tr>
			<td>Iodine for Movat staining</td>
			<td>Sigma-Aldrich, Switzerland</td>
			<td>207772-100G</td>
			<td></td>
		</tr>
		<tr>
			<td>Potassium iodide for Movat pentachrome staining</td>
			<td>Sigma-Aldrich, Switzerland</td>
			<td>60400-100G-F</td>
			<td></td>
		</tr>
		<tr>
			<td>Alcian blue for Movat staining</td>
			<td>Sigma-Aldrich, Switzerland</td>
			<td>A5268-10G</td>
			<td></td>
		</tr>
		<tr>
			<td>Strong Ammonia for Movat pentachrome staining</td>
			<td>Sigma-Aldrich, Switzerland</td>
			<td>320145-500ML</td>
			<td></td>
		</tr>
		<tr>
			<td>Brilliant crocein MOO for Movat pentachrome staining</td>
			<td>Sigma-Aldrich, Switzerland</td>
			<td>210757-50G</td>
			<td></td>
		</tr>
		<tr>
			<td>Acid Fuchsin for Movat pentachrome staining</td>
			<td>Sigma-Aldrich, Switzerland</td>
			<td>F8129-50G</td>
			<td></td>
		</tr>
		<tr>
			<td>Sodium Thiosulfate for Movat pentachrome staining</td>
			<td>Sigma-Aldrich, Switzerland</td>
			<td>72049-250G,</td>
			<td></td>
		</tr>
		<tr>
			<td>Phosphotungstic acid for Movat pentachrome staining</td>
			<td>Sigma-Aldrich, Switzerland</td>
			<td>79690-100G</td>
			<td></td>
		</tr>
		<tr>
			<td>Crocin for Movat pentachrome staining</td>
			<td>Sigma-Aldrich, Switzerland</td>
			<td>17304-5G</td>
			<td></td>
		</tr>
		<tr>
			<td>EUKITT for Movat pentachrome staining</td>
			<td>Sigma-Aldrich, Switzerland</td>
			<td>03989-100ML</td>
			<td></td>
		</tr>
	</tbody>
</table>

the rabbit model of accelerated atherosclerosis: a methodological perspective of the iliac artery balloon injury

Akutes Koronarsyndrom aus koronaren Okklusion nach Entwicklung von atherosklerotischen Plaques und Bruch ist die häufigste Todesursache in der industrialisierten Welt. New Zealand White (NZW) Kaninchen werden häufig als Tiermodell für das Studium der Arteriosklerose verwendet. Sie entwickeln spontanere Läsionen bei mit atherogenen Diät gefüttert; Dies erfordert jedoch langen Zeit von 4 bis 8 Monate. Um weiter zu verbessern und Atherogenese zu beschleunigen, wird häufig eine Kombination von atherogenen Diät und mechanische Endothelzellen Verletzungen eingesetzt. Das vorgestellte Verfahren zur Induktion arteriosklerotische Plaques bei Kaninchen verwendet ein Ballonkatheter, um das Endothel in der linken Beckenkamm Arterie NZW Kaninchen gefüttert mit atherogenen Diät zu stören. Solche mechanischen Schäden, die durch den Ballonkatheter induziert eine Kette von entzündlichen Reaktionen initiieren Neointimal Lipid-Akkumulation in einer Zeit abhängigen Art und Weise. Atherosklerotischen Plaques nach Ballon Verletzungen zeigen Neointimal Verdickung mit umfangreichen Lipid Infiltration, hohe Glattmuskel Zellinhalt und Vorhandensein von Makrophagen abgeleitete Schaumzellen. Diese Technik ist einfach, reproduzierbar und Plaque kontrollierten Länge innerhalb der Beckenkamm Arterie produziert. Das gesamte Verfahren ist innerhalb von 20-30 min abgeschlossen. Das Verfahren ist sicher mit einer niedrigen Mortalität und bietet auch hohen Erfolgsquote bei der Beschaffung von erheblichen Intima Läsionen. Das Verfahren der Ballonkatheter induzierten arteriellen Verletzung führt zu Arteriosklerose innerhalb von zwei Wochen. Dieses Modell eignet sich für die Untersuchung der Krankheit-Pathologie, diagnostische Bildgebung und neue therapeutische Strategien zu bewerten.

Ballon-Verletzung der Beckenkamm Arterie wurde ohne Komplikationen (Abbildung 1) erfolgreich durchgeführt. Die operative Gesamtzeit reichte von 20 bis 30 min für Verletzungen durchgeführt auf nur ein Beckenkamm Arterie und 35 bis 45 min für Verletzungen an beiden Arterien. Die Kaninchen erholte sich innerhalb 1 h nach Ballon-Verletzungen. Alle Tiere erschien ohne signifikanten Gewichtsverlust gesund. Keine Infektion, Ödeme oder arterielle Thrombose aufge...

Watch this Scientific Journal Video about The Rabbit Model of Accelerated Atherosclerosis: A Methodological Perspective of the Iliac Artery Balloon Injury at JoVE.com

The Rabbit Model of Accelerated Atherosclerosis: A Methodological Perspective of the Iliac Artery Balloon Injury

Tiermodelle der Atherosklerose sind notwendig, um den Mechanismus zu verstehen und zu untersuchen, neuere Ansätze zur Vermeidung Plaque Entwicklung oder Bruch, eine führende Ursache des Todes in der industrialisierten Welt. Dieses Protokoll verwendet eine Kombination von Ballon Verletzungen und Cholesterin-reiche Diät, um arteriosklerotische Plaques in Kaninchen Beckenkamm Arterie zu induzieren.

Das Kaninchen-Modell der beschleunigten Atherosklerose: methodischer Sicht der Beckenkamm Arterie Ballon Verletzung

Acute coronary syndrome resulting from coronary occlusion following atherosclerotic plaque development and rupture is the leading cause of death in the ...

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12.0K Aufrufe.  University of Fribourg. Tiermodelle der Atherosklerose sind notwendig, um den Mechanismus zu verstehen und zu untersuchen, neuere Ansätze zur Vermeidung Plaque Entwicklung oder Bruch, eine führende Ursache des Todes in der industrialisierten Welt. Dieses Protokoll verwendet eine Kombination von Ballon Verletzungen und Cholesterin-reiche Diät, um arteriosklerotische Plaques in Kaninchen Beckenkamm Arterie zu induzieren.

Serie: The Rabbit Model of Accelerated Atherosclerosis: A Methodological Perspective of the Iliac Artery Balloon Injury

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 ...

Forschung

Medizin

A Murine Model of Myocardial Ischemia-reperfusion Injury through Ligation of the Left Anterior Descending Artery

Acute or chronic myocardial infarction (MI) are cardiovascular events resulting in high morbidity and mortality. Establishing the pathological mechanisms at work during MI and developing effective therapeutic approaches requires methodology to reproducibly simulate the clinical incidence and reflect the pathophysiological changes associated with MI. Here, we describe a surgical method to induce MI in mouse models that can be used for short-term ischemia-reperfusion (I/R) injury as well as permanent ligation. The major advantage of this method is to facilitate location of the left anterior descending artery (LAD) to allow for accurate ligation of this artery to induce ischemia in the left ventricle of the mouse heart. Accurate positioning of the ligature on the LAD increases reproducibility of infarct size and thus produces more reliable results. Greater precision in placement of the ligature will improve the standard surgical approaches to simulate MI in mice, thus reducing the number of experimental animals necessary for statistically relevant studies and improving our understanding of the mechanisms producing cardiac dysfunction following MI. This mouse model of MI is also useful for the preclinical testing of treatments targeting myocardial damage following MI.

We introduce a surgical method to induce experimental ischemia/reperfusion (I/R) injury to simulate myocardial infarction (MI) in mouse models that allows for more clarity in positioning of the ligation on the left anterior descending artery (LAD) to increase the reproducibility of MI experiments in mice.

Einem Mausmodell der myokardialen Ischämie-Reperfusion Injury durch Ligation des linken vorderen absteigenden Arterie

Acute or chronic myocardial infarction (MI) are cardiovascular events resulting in high morbidity and mortality. Establishing the pathological mechanisms ...

Vascular Balloon Injury and Intraluminal Administration in Rat Carotid Artery

The carotid artery balloon injury model in rats has been well established for over two decades. It remains an important method to study the molecular and cellular mechanisms involved in vascular smooth muscle dedifferentiation, neointima formation and vascular remodeling. Male Sprague-Dawley rats are the most frequently employed animals for this model. Female rats are not preferred as female hormones are protective against vascular diseases and thus introduce a variation into this procedure. The left carotid is typically injured with the right carotid serving as a negative control. Left carotid injury is caused by the inflated balloon that denudes the endothelium and distends the vessel wall. Following injury, potential therapeutic strategies such as the use of pharmacological compounds and either gene or shRNA transfer can be evaluated. Typically for gene or shRNA transfer, the injured section of the vessel lumen is locally transduced for 30 min with viral particles encoding either a protein or shRNA for delivery and expression in the injured vessel wall. Neointimal thickening representing proliferative vascular smooth muscle cells usually peaks at 2 weeks after injury. Vessels are mostly harvested at this time point for cellular and molecular analysis of cell signaling pathways as well as gene and protein expression. Vessels can also be harvested at earlier time points to determine the onset of expression and/or activation of a specific protein or pathway, depending on the experimental aims intended. Vessels can be characterized and evaluated using histological staining, immunohistochemistry, protein/mRNA assays, and activity assays. The intact right carotid artery from the same animal is an ideal internal control. Injury-induced changes in molecular and cellular parameters can be evaluated by comparing the injured artery to the internal right control artery. Likewise, therapeutic modalities can be evaluated by comparing the injured and treated artery to the control injured only artery.

This protocol uses a balloon catheter to cause an intraluminal injury on the rat carotid artery and henceforth elicit neointimal hyperplasia. This is a well-established model for studying the mechanisms of vascular remodeling in response to injury. It is also widely used to determine the validity of potential therapeutic approaches.

Gefäßballon Verletzung und intraluminale Verwaltung in Rattenhalsschlagader

The carotid artery balloon injury model in rats has been well established for over two decades. It remains an important method to study the molecular and ...

A Rat Carotid Balloon Injury Model to Test Anti-vascular Remodeling Therapeutics

The rat carotid balloon injury is a well-established surgical model that has been used to study arterial remodeling and vascular cell proliferation. It is also a valuable model system to test, and to evaluate therapeutics and drugs that negate maladaptive remodeling in the vessel. The injury, or barotrauma, in the vessel lumen caused by an inflated balloon via an inserted catheter induces subsequent neointimal growth, often leading to hyperplasia or thickening of the vessel wall that narrows, or obstructs the lumen. The method described here is sufficiently sensitive, and the results can be obtained in relatively short time (2 weeks after the surgery). The efficacy of the drug or therapeutic against the induced-remodeling can be evaluated either by the post-mortem pathological and histomorphological analysis, or by ultrasound sonography in live animals. In addition, this model system has also been used to determine the therapeutic window or the time course of the administered drug. These studies can leadto the development of a better administrative strategy and a better therapeutic outcome. The procedure described here provides a tool for translational studies that bring drug and therapeutic candidates from bench research to clinical applications.

The rat carotid balloon injury model described below allows researchers to evaluate drugs or therapeutics that negate injury-induced arterial hyperplasia. Detailed pre-surgical preparation, surgical procedure, and post-surgical cares of the animal are described.

Eine Ratte Karotis-Ballon-Verletzung Modell Anti-vaskuläre Remodeling Therapeutics zu Test

The rat carotid balloon injury is a well-established surgical model that has been used to study arterial remodeling and vascular cell proliferation. It is ...

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.

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. ...

In Vivo Gene Transfer to the Rabbit Common Carotid Artery Endothelium

The goal of this method is to introduce a transgene into the endothelium of isolated segments of both rabbit common carotid arteries. The method achieves focal endothelial-selective transgenesis, thereby allowing an investigator to determine the biological roles of endothelial-expressed transgenes and to quantify the in vivo transcriptional activity of DNA sequences in large artery endothelial cells. The method uses surgical isolation of rabbit common carotid arteries and an arteriotomy to deliver a transgene-expressing viral vector into the arterial lumen. A short incubation period of the vector in the lumen, with subsequent aspiration of the lumen contents, is sufficient to achieve efficient and durable expression of the transgene in the endothelium, with no detectable transduction or expression outside of the isolated arterial segment. The method allows assessment of the biological activities of transgene products both in normal arteries and in models of human vascular disease, while avoiding systemic effects that could be caused either by targeting gene delivery to other sites (e.g. the liver) or by the alternative approach of delivering genetic constructs to the endothelium by germ line transgenesis. Application of the method is limited by the need for a skilled surgeon and anesthetist, a well-equipped operating room, the costs of purchasing and housing rabbits, and the need for expertise in gene-transfer vector construction and use. Results obtained with this method include: transgene-related alterations in arterial structure, cellularity, extracellular matrix, or vasomotor function; increases or reductions in arterial inflammation; alterations in vascular cell apoptosis; and progression, retardation, or regression of diseases such as intimal hyperplasia or atherosclerosis. The method also allows measurement of the ability of native and synthetic DNA regulatory sequences to alter transgene expression in endothelial cells, providing results that include: levels of transgene mRNA, levels of transgene protein, and levels of transgene enzymatic activity.

In Vivo Gene Transfer to the Rabbit Common Carotid Artery Endothelium

This method is to introduce a transgene into the endothelium of rabbit carotid arteries. Introduction of the transgene allows the assessment of the biological role of the transgene product either in normal arteries or disease models. The method is also useful for measuring activity of DNA regulatory sequences.

In Vivo Gentransfer in die Kaninchen gemeinsame Halsschlagader Endothel

The goal of this method is to introduce a transgene into the endothelium of isolated segments of both rabbit common carotid arteries. The method achieves ...

A Rabbit Model of Durable Transgene Expression in Jugular Vein to Common Carotid Artery Interposition Grafts

Vein graft bypass surgery is a common treatment for occlusive arterial disease; however, long-term success is limited by graft failure due to thrombosis, intimal hyperplasia, and atherosclerosis. The goal of this article is to demonstrate a method for placing bilateral venous interposition grafts in a rabbit, then transducing the grafts with a gene transfer vector that achieves durable transgene expression. The method allows the investigation of the biological roles of genes and their protein products in normal vein graft homeostasis. It also allows the testing of transgenes for the activities that could prevent vein graft failure, e.g., whether the expression of a transgene prevents the neointimal growth, reduces the vascular inflammation, or reduces atherosclerosis in rabbits fed with a high-fat diet. During an initial survival surgery, the segments of right and left external jugular vein are excised and placed bilaterally as reversed end-to-side common carotid artery interposition grafts. During a second survival surgery, performed 28 days later, each of the grafts is isolated from the circulation with vascular clips and the lumens are filled (via an arteriotomy) with a solution containing a helper-dependent adenoviral (HDAd) vector. After a 20-min incubation, the vector solution is aspirated, the arteriotomy is repaired, and flow is restored. The veins are harvested at time points dictated by individual experimental protocols. The 28-day delay between the graft placement and the transduction is necessary to ensure the adaptation of the vein graft to the arterial circulation. This adaptation avoids rapid loss of transgene expression that occurs in vein grafts transduced before or immediately after grafting. The method is unique in its ability to achieve durable, stable transgene expression in grafted veins. Compared to other large animal vein graft models, rabbits have advantages of low cost and easy handling. Compared to rodent vein graft models, rabbits have larger and easier-to-manipulate blood vessels that provide abundant tissue for analysis.

This method describes the placement of interposition vein grafts in rabbits, the transduction of the grafts, and the achievement of durable transgene expression. This allows the investigation of physiological and pathological roles of transgenes and their protein products in grafted veins, and testing of gene therapies for vein graft disease.

Ein Kaninchen-Modell der dauerhafte Transgene Ausdruck in Halsschlagader, gemeinsame Halsschlagader Interposition Grafts

Vein graft bypass surgery is a common treatment for occlusive arterial disease; however, long-term success is limited by graft failure due to thrombosis, ...

Isometric Contractility Measurement of the Mouse Mesenteric Artery Using Wire Myography

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

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

Messung der isometrischen Kontraktilität der Maus mesenterialen Arterie mit Draht Myographie

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

A Rat Carotid Artery Pressure-Controlled Segmental Balloon Injury with Periadventitial Therapeutic Application

Cardiovascular disease remains the leading cause of death and disability worldwide, in part due to atherosclerosis. Atherosclerotic plaque narrows the luminal surface area in arteries thereby reducing adequate blood flow to organs and distal tissues. Clinically, revascularization procedures such as balloon angioplasty with or without stent placement aim to restore blood flow. Although these procedures reestablish blood flow by reducing plaque burden, they damage the vessel wall, which initiates the arterial healing response. The prolonged healing response causes arterial restenosis, or re-narrowing, ultimately limiting the long-term success of these revascularization procedures. Therefore, preclinical animal models are integral for analyzing the pathophysiological mechanisms driving restenosis, and provide the opportunity to test novel therapeutic strategies. Murine models are cheaper and easier to operate on than large animal models. Balloon or wire injury are the two commonly accepted injury modalities used in murine models. Balloon injury models in particular mimic the clinical angioplasty procedure and cause adequate damage to the artery for the development of restenosis. Herein we describe the surgical details for performing and histologically analyzing the modified, pressure-controlled rat carotid artery balloon injury model. Additionally, this protocol highlights how local periadventitial application of therapeutics can be used to inhibit neointimal hyperplasia. Lastly, we present light sheet fluorescence microscopy as a novel approach for imaging and visualizing the arterial injury in three-dimensions.

The rat carotid artery balloon injury mimics the clinical angioplasty procedure performed to restore blood flow in atherosclerotic vessels. This model induces the arterial injury response by distending the arterial wall, and denuding the intimal layer of endothelial cells, ultimately causing remodeling and an intimal hyperplastic response.

Eine Ratte Carotis Arterie Druck-kontrollierte segmentale Ballonverletzung mit periadventitialen therapeutischen Anwendung

Cardiovascular disease remains the leading cause of death and disability worldwide, in part due to atherosclerosis. Atherosclerotic plaque narrows the ...

The Intra-Aortic Balloon Pump

Cardiogenic shock remains one of the most challenging clinical syndromes in modern medicine. Mechanical support is being increasingly used in the management of cardiogenic shock. Intra-aortic balloon pump (IABP) is one of the earliest and most widely used types of mechanical circulatory support. The device acts by external counterpulsation and uses systolic unloading and diastolic augmentation of aortic pressure to improve hemodynamics. Although IABP provides less hemodynamic support when compared with newer mechanical circulatory support devices, it can still be the mechanical support device of choice in appropriate situations because of its relative simplicity of insertion and removal, need for smaller size vascular access and better safety profile. In this review, we discuss the equipment, procedural and technical aspects, hemodynamic effects, indications, evidence, current status and recent advances in the use of IABP in cardiogenic shock.

We describe the steps for the percutaneous implantation of the intra-aortic balloon pump (IABP), a mechanical circulatory support device. It acts by counterpulsation, inflating at the onset of diastole, augmenting diastolic aortic pressure and improving coronary blood flow and systemic perfusion, and deflating before systole, reducing left ventricular afterload.