Questo lavoro è stato supportato dalla Swiss National Science Foundation Grant 150271.

questo protocollo sperimentale è stato approvato dal ufficio del veterinario cantonale, Friburgo e la Swiss federale veterinaria ufficio, Svizzera (FR 2015/58).
 Nota: conigli di NZW maschio da 2,8 a 3,2 kg sono stati utilizzati. Gli animali sono stati alloggiati in condizioni convenzionali (12 h luce e ciclo scuro, fornito cibo e acqua ad libitum). Prima decumulazione palloncino, gli animali sono stati acclimatati per 1 settimana durante la quale sono stati alimentati con dieta normale. Dopo 1 settimana di acclimatazione, conigli sono stati commutati a dieta atherogenic che consiste di alto contenuto di grassi (8,6%) e acidi grassi saturi con dieta ricca in colesterolo (1%) 205 mg/kg per la durata di studio di tutto. Ferita di palloncino nell'arteria iliaca sinistra è stato effettuato 1 settimana dopo l'inizio della dieta e animali sono stati sacrificati dopo 2 settimane o 4 settimane di infortunio palloncino.
 1. procedure preoperative <ol><li>sterilizzare tutti gli strumenti chirurgici prima dell'uso con uno sterilizzatore del branello di vetro o altro strumento idoneo.</li>
	<li>Preparare e controllare il montaggio del catetere a palloncino.
	<ol><li>Allega un attacco Luer mL 1 siringa riempita con soluzione salina normale per la parte di luer-lock del catetere a palloncino. Monitorare con attenzione per assenza di aria intrappolata. Controllare le perdite e assicurare il gonfiaggio del palloncino corretta premendo lo stantuffo della siringa.</li>
	</ol></li> <li>Pesare il coniglio e accendere il thermopad a 37 ° C.</li>
	<li>Utilizzare una soluzione di buprenorfina ad una concentrazione di 0,3 mg / ml. iniettare una dose di 0,01 mg/kg per via sottocutanea.</li>
	<li>Anesthetize il coniglio con isoflurano 5% e 5 L/min O 2 in una camera di induzione per 10-15 min.</li>
	<li>Posto il coniglio anestetizzato il termocuscino mantenuto sulla piattaforma chirurgica. Inserire la patch e clip per controllare la temperatura, la respirazione e l'elettrocardiogramma.</li>
	<li>Allegare il muso del coniglio ad una maschera di protezione collegata ad una macchina di anestesia adatto. Mantenere l'anestesia con isoflurano (4,0% con 2,5 L/min O 2). Confermare la corretta amputate (indicato dalla mancanza di tono muscolare e perdita dei riflessi gag e padiglioni auricolari).</li>
	<li>Applicare unguento oftalmico in entrambi gli occhi per impedire l'essiccazione le cornee. Coprire il coniglio con un telo chirurgico sterile con solo l'arto inferiore esposto.</li>
</ol> 2. Protocollo chirurgico <ol><li>rimuovere i capelli dalla zona ventrale appena sotto i ginocchi utilizzando tosatrici animali.</li>
	<li>Tampone della zona con un disinfettante adatto per pulire la pelle e rimuovere capelli sciolti.</li>
	<li>Individuare l'arteria safena e fare un'incisione della pelle piccolo di circa 1,5 cm di lunghezza con un bisturi.</li>
	<li>Esporre una piccola porzione dell'arteria safena con piccolo forcipe curvo senza danneggiare la vena femorale e del nervo femorale.</li>
	<li>Posto due cicli di legatura allentata (5-0 seta) sotto l'arteria safena e fascetta una legatura verso l'estremità distale dell'arteria. Posizionare una pinza microvascolare sopra la legatura per fermare il flusso di sangue dall'arteria iliaca.</li>
	<li>Applicare topicamente una goccia di papaverina per dilatare l'arteria e per impedire il vasospasmo.</li>
	<li>Sollevare l'arteria safena con l'aiuto della legatura legata e fare un'incisione piccola arteriotomia utilizzando un 24 ago calibro.</li>
	<li>Elevare i lembi dell'incisione con una pinzetta e inserire lentamente un pick di vena o un ago guida nel lume dell'arteria.</li>
	<li>Inserire un 2 catetere per embolectomia arteriosa Fogarty francese nell'arteria safena. Rimuovere la vena pick e morsetti microvascolari.</li>
	<li>Far avanzare il catetere fino al sesto segno (20-25 cm) corrispondente a una posizione più o meno 2-5 cm di sopra della biforcazione iliaca.</li>
	<li>Gonfiare il palloncino con soluzione fisiologica normale utilizzando una siringa da 1 mL 0,1 mL o ad una pressione nominale di 6 atm utilizzando un regolamentato gonfiatore manuale come descritto in 16 , 22.</li>
	<li>Tenere il catetere a palloncino con il forcipe e trascinare indietro da 6 cm attraverso il arterytoward iliaca il punto di inserimento, durante la rotazione il catetere.</li>
	<li>Sgonfiare il palloncino tirando indietro lo stantuffo della siringa.</li>
	<li>Ripetere i passaggi da 2.10 a 2.13 tre volte per garantire completa decumulazione endoteliale.</li>
	<li>Rimuovere il catetere e legare immediatamente il ciclo di legatura appena sopra il sito di arteriotomia per fermare l'emorragia.</li>
	<li>Applicare antisettico adatto tutto intorno alla periferia della ferita e tampone distanza i coaguli di sangue. Chiudere l'incisione della pelle con una sutura 5 / 0 e disinfettare il sito di chirurgia con povidone-iodio soluzione.</li>
	<li>Ripetere i passaggi da 2.1 a 2.16 su controlaterale iliaca utilizzando un nuovo catetere.</li>
	<li>Tampone l'unguento oftalmico da occhi.</li>
</ol> 3. Cure post-operatorie <ol><li>amministrare sulfadoxine 40 mg/kg e trimetoprim 8 mg/kg o qualsiasi altro antibiotico adatto immediatamente dopo la procedura chirurgica.</li>
	<li>Durante il periodo di anestesia-recupero, mantenere il coniglio sopra un termoforo messo in una gabbia pulita autoclavata.</li>
	<li>Rimuovere la patch monitoraggio e clip.</li>
	<li>Dopo il recupero, tornare i conigli gabbie usate per casa. Iniettare per via sottocutanea buprenorfina 0,05 - 0,1 mg/kg post - operativamente ogni 6-12 h per 48 h. continua dieta atherogenic per altre due settimane o quattro settimane.</li>
</ol> 4. Raccolta di tessuto e analisi della composizione della placca <ol><li>dopo due settimane (per prima placca sottile) o tre settimane di infortunio palloncino, anestetizzare il coniglio utilizzando isoflurano in un modo simile come sopra descritto.</li>
	<li>Aprire la cavità toracica ed eutanasia i conigli dal exsanguination intracardiaca.</li>
	<li>Isolare le arterie iliache, come descritto in 23.
	<ol><li>Brevemente, aprire l'addome ed esporre il retroperitoneum. Traccia l'aorta verso la biforcazione iliaca e legarlo sopra la biforcazione. Rimuovere con cura i tessuti circostanti per esporre e isolare entrambe le arterie iliache.</li>
	</ol></li> <li>Dissezionare fuori entrambe le arterie iliache e immergerli in una soluzione salina tamponata con fosfato ghiacciata. Rimuovere i coaguli con l'aiuto di pinze. Dividere ogni arteria iliaca in 4-6 segmenti per caratterizzare lo spessore della placca in tutta l'arteria.</li>
	<li>Immediatamente incorporare i segmenti arteriosi in uno stampo contenente la temperatura ottimale di taglio composto, snap-congelamento con azoto liquido e tenerlo a-70 ° C. preparare 5 µm spessore sezioni utilizzando un criostato, come descritto in 24.</li>
	L'istologia <li>Perform, immunofluorescenza o immunoistochimica per la morfometria, lipidi della placca e contenuto cellulare come descritto in 10 , 25. 
	Nota: In breve, risciacquare le sezioni arteriose con tamponato fosfato salino (PBS) e permeabilize utilizzando 0,2% Triton. Risciacquare le sezioni con PBS e bloccare i siti non specifici con 2% di sieroalbumina bovina per 30 min. Incubare le sezioni per 1h a 37 ° C con anti actina α-SM (1: 200) o anticorpi RAM11 (1: 200). Risciacquare le sezioni con PBS e li incubare con l'anticorpo secondario appropriato per 30 min a 37 ° C. Lavare nuovamente con PBS e aggiungere Hoechst (5 µ g/mL) per 10 min rilevare nuclei.</li>
</ol>

<ol>
	<li>Mozaffarian, D., 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=Heart+disease+and+stroke+statistics--2015+update:+a+report+from+the+American+Heart+Association.">Heart disease and stroke statistics--2015 update: a report from the American Heart Association.</a> Circulation. 131, e29-e322 (2015).</li><li>Boone, L. R., Brooks, P. A., Niesen, M. I., Ness, G. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mechanism+of+resistance+to+dietary+cholesterol.">Mechanism of resistance to dietary cholesterol.</a> J Lipids. 2011, 101242 (2011).</li><li>Kapourchali, F. R., 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=Animal+models+of+atherosclerosis.">Animal models of atherosclerosis.</a> World J Clin Cases. 2, 126-132 (2014).</li><li>Carter, C. P., Howles, P. N., 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=Genetic+variation+in+cholesterol+absorption+efficiency+among+inbred+strains+of+mice.">Genetic variation in cholesterol absorption efficiency among inbred strains of mice.</a> J Nutr. 127, 1344-1348 (1997).</li><li>Kolodgie, F. D., 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=Hypercholesterolemia+in+the+rabbit+induced+by+feeding+graded+amounts+of+low-level+cholesterol.+Methodological+considerations+regarding+individual+variability+in+response+to+dietary+cholesterol+and+development+of+lesion+type.">Hypercholesterolemia in the rabbit induced by feeding graded amounts of low-level cholesterol. Methodological considerations regarding individual variability in response to dietary cholesterol and development of lesion type.</a> Arterioscler Thromb Vasc Biol. 16, 1454-1464 (1996).</li><li>Singh, V., Tiwari, R. L., Dikshit, M., Barthwal, M. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Models+to+study+atherosclerosis:+a+mechanistic+insight.">Models to study atherosclerosis: a mechanistic insight.</a> Curr Vasc Pharmacol. 7, 75-109 (2009).</li><li>Dornas, W. C., Oliveira, T. T., Augusto, L. E., Nagem, T. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Experimental+atherosclerosis+in+rabbits.">Experimental atherosclerosis in rabbits.</a> Arq Bras Cardiol. 95, 272-278 (2010).</li><li>Baumgartner, H. R., Studer, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=[Effects+of+vascular+catheterization+in+normo-+and+hypercholesteremic+rabbits].">[Effects of vascular catheterization in normo- and hypercholesteremic rabbits].</a> Pathol Microbiol (Basel). 29, 393-405 (1966).</li><li>Tanaka, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sustained+activation+of+vascular+cells+and+leukocytes+in+the+rabbit+aorta+after+balloon+injury.">Sustained activation of vascular cells and leukocytes in the rabbit aorta after balloon injury.</a> Circulation. 88, 1788-1803 (1993).</li><li>Phinikaridou, A., Hallock, K. J., Qiao, Y., Hamilton, J. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+robust+rabbit+model+of+human+atherosclerosis+and+atherothrombosis.">A robust rabbit model of human atherosclerosis and atherothrombosis.</a> J Lipid Res. 50, 787-797 (2009).</li><li>Nakazawa, 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=Drug-eluting+stent+safety:+findings+from+preclinical+studies.">Drug-eluting stent safety: findings from preclinical studies.</a> Expert Rev Cardiovasc Ther. 6, 1379-1391 (2008).</li><li>Aikawa, 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=Lipid+lowering+by+diet+reduces+matrix+metalloproteinase+activity+and+increases+collagen+content+of+rabbit+atheroma:+a+potential+mechanism+of+lesion+stabilization.">Lipid lowering by diet reduces matrix metalloproteinase activity and increases collagen content of rabbit atheroma: a potential mechanism of lesion stabilization.</a> Circulation. 97, 2433-2444 (1998).</li><li>Jeanpierre, E., 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=Dietary+lipid+lowering+modifies+plaque+phenotype+in+rabbit+atheroma+after+angioplasty:+a+potential+role+of+tissue+factor.">Dietary lipid lowering modifies plaque phenotype in rabbit atheroma after angioplasty: a potential role of tissue factor.</a> Circulation. 108, 1740-1745 (2003).</li><li>Durand, E., 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=Magnetic+resonance+imaging+of+ruptured+plaques+in+the+rabbit+with+ultrasmall+superparamagnetic+particles+of+iron+oxide.">Magnetic resonance imaging of ruptured plaques in the rabbit with ultrasmall superparamagnetic particles of iron oxide.</a> J Vasc Res. 44, 119-128 (2007).</li><li>Stadius, M. L., 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=Time+course+and+cellular+characteristics+of+the+iliac+artery+response+to+acute+balloon+injury.+An+angiographic,+morphometric,+and+immunocytochemical+analysis+in+the+cholesterol-fed+New+Zealand+white+rabbit.">Time course and cellular characteristics of the iliac artery response to acute balloon injury. An angiographic, morphometric, and immunocytochemical analysis in the cholesterol-fed New Zealand white rabbit.</a> Arterioscler Thromb. 12, 1267-1273 (1992).</li><li>Khanna, V., 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=Cholesterol+diet+withdrawal+leads+to+an+initial+plaque+instability+and+subsequent+regression+of+accelerated+iliac+artery+atherosclerosis+in+rabbits.">Cholesterol diet withdrawal leads to an initial plaque instability and subsequent regression of accelerated iliac artery atherosclerosis in rabbits.</a> PLoS One. 8, e77037 (2013).</li><li>Zou, J., 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=Effect+of+resveratrol+on+intimal+hyperplasia+after+endothelial+denudation+in+an+experimental+rabbit+model.">Effect of resveratrol on intimal hyperplasia after endothelial denudation in an experimental rabbit model.</a> Life Sci. 68, 153-163 (2000).</li><li>Li, M., Zhang, Y., Ren, H., Zhang, Y., Zhu, X. <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+clopidogrel+on+the+inflammatory+progression+of+early+atherosclerosis+in+rabbits+model.">Effect of clopidogrel on the inflammatory progression of early atherosclerosis in rabbits model.</a> Atherosclerosis. 194, 348-356 (2007).</li><li>Nakazawa, 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=Evaluation+of+polymer-based+comparator+drug-eluting+stents+using+a+rabbit+model+of+iliac+artery+atherosclerosis.">Evaluation of polymer-based comparator drug-eluting stents using a rabbit model of iliac artery atherosclerosis.</a> Circ Cardiovasc Interv. 4, 38-46 (2011).</li><li>Van Dyck, C. J., 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=Resolute+and+Xience+V+polymer-based+drug-eluting+stents+compared+in+an+atherosclerotic+rabbit+double+injury+model.">Resolute and Xience V polymer-based drug-eluting stents compared in an atherosclerotic rabbit double injury model.</a> Catheter Cardiovasc Interv. 81, E259-E268 (2013).</li><li>Abran, 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=Validating+a+bimodal+intravascular+ultrasound+(IVUS)+and+near-infrared+fluorescence+(NIRF)+catheter+for+atherosclerotic+plaque+detection+in+rabbits.">Validating a bimodal intravascular ultrasound (IVUS) and near-infrared fluorescence (NIRF) catheter for atherosclerotic plaque detection in rabbits.</a> Biomed Opt Express. 6, 3989-3999 (2015).</li><li>Kanamasa, K., 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=Recombinant+tissue+plasminogen+activator+prevents+intimal+hyperplasia+after+balloon+angioplasty+in+hypercholesterolemic+rabbits.">Recombinant tissue plasminogen activator prevents intimal hyperplasia after balloon angioplasty in hypercholesterolemic rabbits.</a> Jpn Circ J. 60, 889-894 (1996).</li><li>Pai, 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=Inhibition+of+in-stent+restenosis+in+rabbit+iliac+arteries+with+photodynamic+therapy.">Inhibition of in-stent restenosis in rabbit iliac arteries with photodynamic therapy.</a> Eur J Vasc Endovasc Surg. 30, 573-581 (2005).</li><li>Fischer, A. H., Jacobson, K. A., Rose, J., Zeller, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cryosectioning+tissues.">Cryosectioning tissues.</a> CSH Protoc. 2008, (2008).</li><li>Chaytor, A. T., Bakker, L. M., Edwards, D. H., Griffith, T. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Connexin-mimetic+peptides+dissociate+electrotonic+EDHF-type+signalling+via+myoendothelial+and+smooth+muscle+gap+junctions+in+the+rabbit+iliac+artery.">Connexin-mimetic peptides dissociate electrotonic EDHF-type signalling via myoendothelial and smooth muscle gap junctions in the rabbit iliac artery.</a> Br J Pharmacol. 144, 108-114 (2005).</li><li>Zhang, W., Trebak, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Vascular+balloon+injury+and+intraluminal+administration+in+rat+carotid+artery.">Vascular balloon injury and intraluminal administration in rat carotid artery.</a> J Vis Exp. (94), (2014).</li><li>Maillard, L., 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=Effect+of+percutaneous+adenovirus-mediated+Gax+gene+delivery+to+the+arterial+wall+in+double-injured+atheromatous+stented+rabbit+iliac+arteries.">Effect of percutaneous adenovirus-mediated Gax gene delivery to the arterial wall in double-injured atheromatous stented rabbit iliac arteries.</a> Gene Ther. 7, 1353-1361 (2000).</li><li>Sharif, F., 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=Gene-eluting+stents:+adenovirus-mediated+delivery+of+eNOS+to+the+blood+vessel+wall+accelerates+re-endothelialization+and+inhibits+restenosis.">Gene-eluting stents: adenovirus-mediated delivery of eNOS to the blood vessel wall accelerates re-endothelialization and inhibits restenosis.</a> Mol Ther. 16, 1674-1680 (2008).</li><li>Lee, J. 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=Development+of+a+rabbit+model+for+a+preclinical+comparison+of+coronary+stent+types+in-vivo.">Development of a rabbit model for a preclinical comparison of coronary stent types in-vivo.</a> Korean Circ J. 43, 713-722 (2013).</li><li>Tulis, D. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rat+carotid+artery+balloon+injury+model.">Rat carotid artery balloon injury model.</a> Methods Mol Med. 139, 1-30 (2007).</li><li>Asada, Y., 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=Effects+of+inflation+pressure+of+balloon+catheter+on+vascular+injuries+and+subsequent+development+of+intimal+hyperplasia+in+rabbit+aorta.">Effects of inflation pressure of balloon catheter on vascular injuries and subsequent development of intimal hyperplasia in rabbit aorta.</a> Atherosclerosis. 121, 45-53 (1996).</li><li>Dornas, W. C., Oliveira, T. T., Augusto, L. E., Nagem, T. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Experimental+atherosclerosis+in+rabbits.">Experimental atherosclerosis in rabbits.</a> Arq Bras Cardiol. 95, 272-278 (2010).</li><li>Waksman, R., 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=PhotoPoint+photodynamic+therapy+promotes+stabilization+of+atherosclerotic+plaques+and+inhibits+plaque+progression.">PhotoPoint photodynamic therapy promotes stabilization of atherosclerotic plaques and inhibits plaque progression.</a> J Am Coll Cardiol. 52, 1024-1032 (2008).</li><li>Fernandez-Parra, R., 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=Pharmacokinetic+Study+of+Paclitaxel+Concentration+after+Drug-Eluting+Balloon+Angioplasty+in+the+Iliac+Artery+of+Healthy+and+Atherosclerotic+Rabbit+Models.">Pharmacokinetic Study of Paclitaxel Concentration after Drug-Eluting Balloon Angioplasty in the Iliac Artery of Healthy and Atherosclerotic Rabbit Models.</a> J Vasc Interv Radiol. 26, 1380-1387 (2015).</li><li>Dussault, S., Dhahri, W., Desjarlais, M., Mathieu, R., Rivard, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Elsibucol+inhibits+atherosclerosis+following+arterial+injury:+multifunctional+effects+on+cholesterol+levels,+oxidative+stress+and+inflammation.">Elsibucol inhibits atherosclerosis following arterial injury: multifunctional effects on cholesterol levels, oxidative stress and inflammation.</a> Atherosclerosis. 237, 194-199 (2014).</li><li>Manderson, J. 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>

Gli autori non dichiarano concorrenti interessi finanziari.

Il modello di aterosclerosi dell'arteria iliaca di coniglio è ampiamente usato nella ricerca di aterosclerosi. Con questo protocollo i conigli sviluppato rapidamente placche più gravi e avanzate rispetto alle lesioni spontanee sviluppate con solo dieta ricca in colesterolo. D'importanza, gli animali recuperano rapidamente dalla chirurgia.
Lo stimolo principale per atherogenesis è i danni meccanici causati dal catetere a palloncino che danneggia l'endotelio e si distende il vaso muro<sup cla...

La rottura delle placche aterosclerotiche vulnerabili è una delle principali cause di morte nei paesi industrializzati1. Anche se la ricerca negli ultimi decenni ha spiegato diversi meccanismi cellulari e molecolari coinvolti nella progressione della placca, ha continuato gli sforzi sono ancora necessari non solo per svelare il complesso meccanismo della progressione di malattia ma anche per sperimentare nuove terapeutico si avvicina. Sono stati proposti diversi modelli animali per studiare l'aterosclerosi. Manipolazione genetica, lesioni di endotelio di alimentazione o meccanico di colesterolo sono le strategie standard condivise da modelli più animali di aterosclerosi tra cui topi, conigli o cavie. Tra questi, conigli di NZW sono sensibili alla dieta colesterolo mentre topi e ratti normali non assorbono significativamente colesterolo dietetico2,3,4. Conigli sviluppano spontaneamente lesioni aortiche ricche di macrofagi con qualche componente fibrosa quando alimentati con dieta ricca di colesterolo5,6. Tuttavia, il tempo lungo preparatorio di 4-8 mesi per indurre aterosclerotica plaquesby alimentazione colesterolo dieta da solo6,7 è un grande svantaggio per la maggior parte delle impostazioni sperimentali. Nell'inseguimento per indurre le lesioni in tempi relativamente brevi, una combinazione di colesterolo alto dieta e palloncino ferita è stata sviluppata da Baumgarter e Studer8. L'obiettivo generale di questa tecnica è quello di indurre le placche aterosclerotiche, composte di cellule schiumose (simile a striscia grassa in esseri umani) in conigli ipercolesterolemici entro 2 settimane. La tecnica attuale descrive la procedura di lesione della parete arteriosa basato sul metodo di Baumgarter utilizzando un catetere a palloncino avanzato nell'arteria iliaca di conigli ipercolesterolemici NZW.
Insieme ad una dieta ricca di colesterolo, lesioni risultanti da palloncino indotto de-endotelizzazione porterà ad aterosclerosi. Lesioni palloncino accelera la formazione di lesioni aterosclerotiche e produce la placca di dimensione uniforme e distribuzione. L'ispessimento intimal aumenta per un periodo di tempo e inizia l'infiltrazione delle cellule intimal entro pochi giorni dopo la lesione. Strisce grasse con sostanziale macrofagi iniziano a comparire dopo 7-10 giorni della ferita dell'aerostato e sono rappresentate come lesione di tipo II secondo la classificazione di associazione americana del cuore. Ferita di palloncino in coniglio è spesso eseguita in aorta a studiare composizione della placca. L'endotelio neointimale esprime elevati livelli della molecola di adesione intercellulare. Le placche sono associate con la dissezione mediale e modifiche adventitial. Le lesioni aterosclerotiche sono costituite da lipidi, cellule di proliferazione del muscolo liscio (SMCs), fibre di collagene e cellule infiammatorie che si accumulano sotto l'endotelio rigenerata e sono per lo più di tipo II in natura. La distribuzione topologica delle placche di coniglio era simile a quella riportata in aorte umano 9,10 In linea di principio, l'aorta è più grande nelle dimensioni rispetto alle arterie iliache e produrrebbe placca di lunghezza maggiore. Tuttavia, il principale vantaggio di usando l'arteria iliaca come il sito di aterosclerosi in conigli è la sua accessibilità, la somiglianza nel contenuto muscolare umane dell'arteria coronaria11, lesione uniforme sviluppo12, fattore del tessuto ad alta attività13 e coerente nave dimensione paragonabile a umane dell'arteria coronaria, permettendo la valutazione dei dispositivi commercialmente fabbricati a morfometriche ed angiografici endpoint. Metodi invasivi e non invasivi sono stati studiati per analizzare le placche nelle arterie iliache coniglio in animale vivo. Rapporti precedenti descrivono l'uso della risonanza magnetica per immagini (MRI) con l'aiuto di un sistema di MR 2,35-tesla 14 inoltre, ultrasuoni intravascolari (IVUS) o cateteri di tomografia a coerenza ottica possono essere opportunamente applicata all'immagine placche aterosclerotiche nelle arterie iliache coniglio. L'arteria iliaca è accessibile per l'imaging ad ultrasuoni quando si utilizza un'ecografia ad alta risoluzione e l'aorta può essere esplorata anche con questa tecnica.
Nell'ultimo decennio, questo modello di coniglio della lesione palloncino ha aiutato a capire meglio i meccanismi della placca progressione15e placca regressione16. Inoltre, il modello è stato utilizzato per studiare l'influenza di nuovi agenti terapeutici quali le statine, gli agenti antipiastrinici standard, antiossidante agenti17,18 e stent medicati come everolimus o zotarolimus-eluting stent19,20 sull'ispessimento neointimal. Questo modello è stato utilizzato anche per indagare imaging intravascolare del catetere21di imaging di fluorescenza di vicino infrarosso.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<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>
		<tr>
			<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>
		</tr>
		<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>
		</tr>
		<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

Sindrome coronarica acuta derivando dall'occlusione coronaria dopo la rottura e lo sviluppo della placca aterosclerotica è la principale causa di morte nel mondo industrializzato. Conigli di bianco di Nuova Zelanda (NZW) sono ampiamente usati come modello animale per lo studio dell'aterosclerosi. Si sviluppano lesioni spontanee quando alimentati con dieta atherogenic; Tuttavia, questo richiede molto tempo di 4-8 mesi. Per ulteriormente migliorare e accelerare l'aterogenesi, una combinazione di dieta atherogenic e lesione endoteliale meccanica è spesso impiegata. La procedura presentata per indurre le placche aterosclerotiche in conigli utilizza un catetere a palloncino di perturbare l'endotelio nell'arteria iliaca sinistra di NZW conigli alimentati con dieta atherogenic. Tali danni meccanici causati dal catetere a palloncino induce una catena di reazioni infiammatorie neointimale accumulo di lipidi in un modo dipendente dal tempo di avvio. La placca aterosclerotica seguendo palloncino lesioni Visualizza neointimale l'ispessimento con infiltrazione lipidica vasto, contenuto della cella di muscolo liscio alta e presenza di macrofagi derivati cellule della gomma piuma. Questa tecnica è semplice, riproducibile e produce placca di lunghezza controllata all'interno dell'arteria iliaca. Tutta la procedura è completata entro 20-30 min. La procedura è sicura con bassa mortalità e offre anche elevato successo nell'ottenere notevoli lesioni intimale. La procedura del catetere a palloncino indotto risultati di lesione arteriosa nell'aterosclerosi entro due settimane. Questo modello può essere utilizzato per indagare la patologia di malattia, diagnostica per immagini e valutare nuove strategie terapeutiche.

Lesioni dell'aerostato dell'arteria iliaca è stata effettuata con successo senza complicazione (Figura 1). Il tempo operatorio totale variato da 20 a 30 min per lesioni eseguite su soltanto un'arteria iliaca e 35 a 45 min per lesioni su entrambe le arterie. Il coniglio ha recuperato entro 1 h dopo la ferita di palloncino. Tutti gli animali è apparso sani senza perdita di peso significativa. È stata rilevata alcuna infezione, edema o trombosi arteriosa. Del...

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

Modelli animali di aterosclerosi sono essenziali per capire il meccanismo e per indagare più recenti approcci per prevenire lo sviluppo della placca o rottura, delle cause principali di morte nel mondo industrializzato. Questo protocollo utilizza una combinazione di lesioni palloncino e dieta ricca di colesterolo per indurre placche aterosclerotiche nell'arteria iliaca coniglio.

Il modello del coniglio di aterosclerosi accelerata: una prospettiva metodologica della ferita dell'aerostato dell'arteria iliaca

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 Views.  University of Fribourg. Modelli animali di aterosclerosi sono essenziali per capire il meccanismo e per indagare più recenti approcci per prevenire lo sviluppo della placca o rottura, delle cause principali di morte nel mondo industrializzato. Questo protocollo utilizza una combinazione di lesioni palloncino e dieta ricca di colesterolo per indurre placche aterosclerotiche nell'arteria iliaca coniglio.

Video: 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.

Un modello murino di impianto di stent a livello dell&#39;arteria carotidea per lo Studio della restenosi

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

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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.

Un modello murino di ischemia miocardica e riperfusione lesioni da legatura della arteria discendente anteriore sinistra

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.

Vascolare Balloon Lesioni e intraluminale Amministrazione in Rat carotide

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.

Un ratto carotidea Pallone Lesioni modello per verificare Therapeutics ritocca Anti-vascolari

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.

Induzione di aterosclerosi accelerata nei topi: il modello "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. ...

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 Trasferimento del gene per l'endotelio dell'arteria carotica comune di coniglio

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.

Un modello del coniglio di espressione del Transgene durevole nella vena giugulare di innesti di interposizione dell'arteria carotica comune

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.

Misurazione di contrattilità isometrica dell'arteria mesenterica Mouse con filo Myography

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.

Una lesione segmentale del palloncino segmentale controllato dalla pressione carotide del ratto con applicazione terapeutica periadventitiale

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.