Name: a rat carotid balloon injury model to test anti-vascular remodeling therapeutics
Uploaded: 2016-09-19T12:30:00
Description: Watch this Scientific Journal Video about A Rat Carotid Balloon Injury Model to Test Anti-vascular Remodeling Therapeutics at JoVE.com

The work was supported by the intramural research program of the NIH, National Institute on Aging, and by a Priority Research Centers Program grant from the National Research Foundation (NRF-2009-0093812) funded by the Ministry of Science, Information and Communication Technology, &amp; Future Planning, the Republic of Korea (H.T.). We thank Dr. Han-sol Park for putting "material" part together for the manuscript.

NOT: sıçan balon yaralanması modelinde ve rekombinant sRAGE ve ultrason ultrason çalışmaların enjeksiyonu da dahil olmak üzere ilgili prosedürlerin kullanılması Ulusal Yaşlanma Enstitüsü Hayvan Bakım ve Kullanım Komitesi (ACUC), NIH tarafından onaylanmıştır. 1. Ön cerrahi Hazırlıklar <ol><li> Enstrüman, Cerrahi Platformu ve Kişisel Koruma Ekipmanı. <ol><li> Bu prosedürde kullanılan tüm cerrahi aletler ve reaktifler için malzeme ve ekip...

<ol>
	<li>Chaabane, C., Otsuka, F., Virmani, R., Bochaton-Piallat, M. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Biological+responses+in+stented+arteries.">Biological responses in stented arteries.</a> Cardiovasc Res. 99, 353-363 (2013).</li><li>Goel, S. A., Guo, L. W., Liu, B., Kent, K. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mechanisms+of+post-intervention+arterial+remodelling.">Mechanisms of post-intervention arterial remodelling.</a> Cardiovasc Res. 96, 363-371 (2012).</li><li>Khan, R., Agrotis, A., Bobik, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Understanding+the+role+of+transforming+growth+factor-beta1+in+intimal+thickening+after+vascular+injury.">Understanding the role of transforming growth factor-beta1 in intimal thickening after vascular injury.</a> Cardiovasc Res. 74, 223-234 (2007).</li><li>Schillinger, M., Minar, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Restenosis+after+percutaneous+angioplasty:+the+role+of+vascular+inflammation.">Restenosis after percutaneous angioplasty: the role of vascular inflammation.</a> Vasc Health Risk Manag. 1, 73-78 (2005).</li><li>Katz, G., Harchandani, B., Shah, B. <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+stents:+the+past,+present,+and+future.">Drug-eluting stents: the past, present, and future.</a> Curr Atheroscler Rep. 17, 485 (2015).</li><li>Jain, M., Singh, A., Singh, V., 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=Involvement+of+Interleukin-1+Receptor-Associated+Kinase-1+in+Vascular+Smooth+Muscle+Cell+Proliferation+and+Neointimal+Formation+After+Rat+Carotid+Injury.">Involvement of Interleukin-1 Receptor-Associated Kinase-1 in Vascular Smooth Muscle Cell Proliferation and Neointimal Formation After Rat Carotid Injury.</a> Arterioscler Thromb Vasc Biol. , (2015).</li><li>Lee, K. P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Carvacrol+inhibits+atherosclerotic+neointima+formation+by+downregulating+reactive+oxygen+species+production+in+vascular+smooth+muscle+cells.">Carvacrol inhibits atherosclerotic neointima formation by downregulating reactive oxygen species production in vascular smooth muscle cells.</a> Atherosclerosis. 240, 367-373 (2015).</li><li>Li, G., Chen, S. J., Oparil, S., Chen, Y. F., Thompson, 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=Direct+in+vivo+evidence+demonstrating+neointimal+migration+of+adventitial+fibroblasts+after+balloon+injury+of+rat+carotid+arteries.">Direct in vivo evidence demonstrating neointimal migration of adventitial fibroblasts after balloon injury of rat carotid arteries.</a> Circulation. 101 (12), 1362-1365 (2000).</li><li>Noda, 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=New+endoplasmic+reticulum+stress+regulator,+gipie,+regulates+the+survival+of+vascular+smooth+muscle+cells+and+the+neointima+formation+after+vascular+injury.">New endoplasmic reticulum stress regulator, gipie, regulates the survival of vascular smooth muscle cells and the neointima formation after vascular injury.</a> Arterioscler Thromb Vasc Biol. 35 (5), 1246-1253 (2015).</li><li>Deuse, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dichloroacetate+prevents+restenosis+in+preclinical+animal+models+of+vessel+injury.">Dichloroacetate prevents restenosis in preclinical animal models of vessel injury.</a> Nature. 509 (7502), 641-644 (2014).</li><li>Guo, 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=p55gamma+functional+mimetic+peptide+N24+blocks+vascular+proliferative+disorders.">p55gamma functional mimetic peptide N24 blocks vascular proliferative disorders.</a> J Mol Med (Berl). , (2015).</li><li>Oh, 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=Dimethylfumarate+attenuates+restenosis+after+acute+vascular+injury+by+cell-specific+and+Nrf2-dependent+mechanisms.">Dimethylfumarate attenuates restenosis after acute vascular injury by cell-specific and Nrf2-dependent mechanisms.</a> Redox Biol. 2, 855-864 (2014).</li><li>Tae, H. 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=The+N-glycoform+of+sRAGE+is+the+key+determinant+for+its+therapeutic+efficacy+to+attenuate+injury-elicited+arterial+inflammation+and+neointimal+growth.">The N-glycoform of sRAGE is the key determinant for its therapeutic efficacy to attenuate injury-elicited arterial inflammation and neointimal growth.</a> J Mol Med (Berl. 91 (12), 1369-1381 (2013).</li><li>Zhou, Z., 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=Receptor+for+AGE+(RAGE)+mediates+neointimal+formation+in+response+to+arterial+injury.">Receptor for AGE (RAGE) mediates neointimal formation in response to arterial injury.</a> Circulation. 107 (17), 2238-2243 (2003).</li><li>Lindner, V., Fingerle, J., Reidy, M. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mouse+model+of+arterial+injury.">Mouse model of arterial injury.</a> Circ Res. 73 (5), 792-796 (1993).</li><li>Le, V., Johnson, C. G., Lee, J. D., Baker, A. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Murine+model+of+femoral+artery+wire+injury+with+implantation+of+a+perivascular+drug+delivery+patch.">Murine model of femoral artery wire injury with implantation of a perivascular drug delivery patch.</a> J Vis Exp. (96), e52403 (2015).</li><li>Iannaccone, P. M., Jacob, H. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rats!+.">Rats! .</a> Dis Model Mech. 2 (5-6), 206-210 (2009).</li><li>Fu, Y., Sander, J. D., Reyon, D., Cascio, V. M., Joung, J. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Improving+CRISPR-Cas+nuclease+specificity+using+truncated+guide+RNAs.">Improving CRISPR-Cas nuclease specificity using truncated guide RNAs.</a> Nat Biotechnol. 32 (3), 279-284 (2014).</li><li>Sander, J. D., Joung, J. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=CRISPR-Cas+systems+for+editing,+regulating+and+targeting+genomes.">CRISPR-Cas systems for editing, regulating and targeting genomes.</a> Nat Biotechnol. 32 (4), 347-355 (2014).</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>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>Tae, H. 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=Vessel+ultrasound+sonographic+assessment+of+soluble+receptor+for+advanced+glycation+end+products+efficacy+in+a+rat+balloon+injury+model.">Vessel ultrasound sonographic assessment of soluble receptor for advanced glycation end products efficacy in a rat balloon injury model.</a> Curr Ther Res Clin Exp. 76, 110-115 (2014).</li><li>Tae, H. 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=Chronic+treatment+with+a+broad-spectrum+metalloproteinase+inhibitor,+doxycycline,+prevents+the+development+of+spontaneous+aortic+lesions+in+a+mouse+model+of+vascular+Ehlers-Danlos+syndrome.">Chronic treatment with a broad-spectrum metalloproteinase inhibitor, doxycycline, prevents the development of spontaneous aortic lesions in a mouse model of vascular Ehlers-Danlos syndrome.</a> J Pharmacol Exp Ther. 343 (1), 246-251 (2012).</li><li>Sakaguchi, 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=Central+role+of+RAGE-dependent+neointimal+expansion+in+arterial+restenosis.">Central role of RAGE-dependent neointimal expansion in arterial restenosis.</a> J Clin Invest. 111 (7), 959-972 (2003).</li></ol>

karotid arter lümeninde duvar endoteli kaldırır yaralanma oluşturmak için balonu şişirmek için kullanılan iki yöntem olmuştur. Bir sıvı 20 ile bağlı şırıngayı doldurmak ve diğer hava basıncı 21 kullanmaktır. Biz tam sıvı hacmi (0.02 ml) her işlem için kullanılacak, çünkü sıvı dolu şırıngayı kullanmayı tercih eder. Bu prosedür uygulanan her bir hayvanın yaralanma benzer bir seviyeye neden balon bir hassas ve tekrarlanabilir enflasyonu oluşturur. şırınga s?...

Anjiyoplasti ateroskleroz gibi patolojik durumlar sonuçtaki daralmış veya tıkalı damarları genişletmek için kullanılan bir endovasküler işlemdir. anjiyoplasti yaygın bir komplikasyon cerrahi yaralanmalar ve sonraki inflamasyon kaynaklı vasküler yeniden nedeniyle oluşur ameliyat sonrası neointimal hiperplazi veya restenoz vardır. Bu koşullar damar düz hücreler ve çoklu patofizyolojik 1-3 çoğalmasına yol açar. Neointimal hiperplazi gemi yeniden kalınlaşır ve ilk yıl içinde% 60 anjiyoplasti sonrası hastalarda kadar oluşur. Bu nedenle, restenoz yaygın olarak kullanılan anjiyoplasti prosedür 4 büyük bir başarısızlık. Uyuşturucu elüsyon stent implantasyonu restenoz engellemeye yardımcı olsa da, yalnızca seçilen adaylar bu pahalı prosedürü 5 uğrayabilir. hayvan ve klinik çalışmalar hem kronik inflamasyon vascu tarafından oluşturulan kurduklar yaralanma ve / veya cerrahi yara anjiyoplasti sonrası neointimal büyüme 2,4 için ana uyarıcı olarak hizmet vermektedir. Bu nedenle sıçan karotis balon yaralanma modeli taklit klinik durum ve vasküler yeniden yapılanma ve vasküler hücre çoğalması 6-9 katıldıklarını hücresel faktörleri belirlemek için değerli bir model sistem olarak hizmet vermektedir. Bu model sistemi, ön-klinik çalışmalarda translasyonel 10-14 neointimal büyümede ilaçlarla tedavi ve tedavi reaktifler için son derece yararlıdır değerlendirmek için bir araç ve / veya ekran. Fare karotis tel yaralanması modelinde 15 ve fare femoral arter tel yaralanması modelinde 16, sıçan karotis balon yaralanma modeli verdirdiler yaralanma tekrarlanabilirlik kolaylaştırır cerrahi prosedür kolaylığı için boyutu yeterince büyük olma avantajları vardır karşılaştırıldığında. Bu addit edecek birincil hücreler daha çok sayıda (örneğin, vasküler düz kas hücreleri, endotelyal hücreler) sağlayabilirional in vitro çalışmalar vasküler remodeling düzenleyen moleküler mekanizmayı tanımlamak için. Önemli olarak, farelere kıyasla, sıçanlar fizyolojik ve toksikolojik çalışmalar 17 için daha iyi bir model olduğu bilinmektedir. Sıçan modeli bir dezavantaj ya da sınırlama genetik modifiye ve gen nakavt modellerinin olmaması olmasına rağmen, bu dezavantaj sıçan genomik dizisinin kullanılabilirliği ve bu tür işler CRISPR-CAS teknolojisi gibi güçlü genomik düzenleme araçları son gelişme ile aşılabilir farklı model sistemlerde 18,19 genomik dizilerin geniş aralıkları mümkün manipülasyon. Sıçan balon yaralanma modeli birden laboratuarları ve çeşitli kapsamlı protokoller tarafından kullanılmasına rağmen 20,21 yayınlanmış, bu protokol öncesi cerrahi hazırlıkları daha fazla ayrıntı vermeyi amaçlamaktadır ve bu cerrahi uygulama kurmak için bu prosedüre yeni araştırmacılara yol açabilir. Biz de t ameliyat sonrası bakım vurgulamakCanlı hayvanların 13,22 değil sadece arteriyel remodeling terapötik etki postmortem patolojik ve histomorfolojik analizler değil, aynı zamanda ultrason sonografik çalışmalar veriyor o hayvanlar.

<table border="0" cellpadding="0" cellspacing="0" width="450">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:287px;">2 F Fogarty balloon embolectomy catheter&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;</td>
			<td style="width:163px;">Edwards Lifesciences&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:287px;">Standard scalpel</td>
			<td style="width:163px;">Fine Science Tools</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:287px;">Small&#160; curved forceps (Large radius Dumont#7shanks curved)&#160;&#160;&#160;&#160;&#160;&#160;&#160;</td>
			<td style="width:163px;">Fine Science Tools&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:287px;">Large, medium and small micro-scissors</td>
			<td style="width:163px;">Roboz</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:287px;">Needles (20 G)&#160;&#160;</td>
			<td style="width:163px;">TycoHealthcare</td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:287px;">Micro-surgery forceps with micro-blunted atraumatic tips</td>
			<td style="width:163px;">Fine Science Tools&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:287px;">Atraumatic straight small arterial clamps&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;</td>
			<td style="width:163px;">Fine Science Tools&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;</td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:287px;">Retractor&#160; with maximum spread 5.5 cm long blunt teeth</td>
			<td style="width:163px;">Fine Science Tools&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:287px;">Silk suture (4.0 and&#160; 6.0 )</td>
			<td style="width:163px;">Fine Science Tools&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:287px;">Syringe (1.0 ml)&#160;</td>
			<td style="width:163px;">BD&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:287px;">Curity gauze sponges</td>
			<td style="width:163px;">AllegroMedical</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:287px;">Cotton tip applicators sterile and non-sterile</td>
			<td style="width:163px;">Puritan Medical Products</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:287px;">Compact hot bead sterilizer</td>
			<td style="width:163px;">Fine Science Tools</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:287px;">Self-regulating heating pad</td>
			<td style="width:163px;">Fine Science Tools&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:287px;">ADS200 anesthesia system/ventilator</td>
			<td style="width:163px;">Paragon Medical</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:287px;">Isoflurane (forane), liquid form</td>
			<td style="width:163px;">Baxter</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:287px;">Sodium chloride 0.9% (Saline)&#160;</td>
			<td style="width:163px;">Hospira</td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:287px;">Buprenex (buprenorphine)&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;</td>
			<td style="width:163px;">Reckitt Benckiser Healthcare (UK) Ltd.&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:287px;">70% alcohol</td>
			<td style="width:163px;">Fisher</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:287px;">1: 10 Betadine</td>
			<td style="width:163px;">Fisher</td>
		</tr>
	</tbody>
</table>

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.

İki hafta balon yaralanma sonrası, sıçan ötenazi ve karotis arterler histo-morfolojik analizler için izole edilmiştir. Hem sol karotis arter ve un sağ işletilen arter işlenmiş, kesit çapraz ve parafine gömülü olduğunu işletilmektedir. parafin numuneleri daha sonra ince kesitli ve hematoksilen-eozin (H &amp; E) ile boyandı vardır. Histo-Şekil incelemesi dijital görüntü analiz sistemi kullanılarak gerçekleştirilir. Arter hasat ve Histo-morfolojik analiz ayrıntıl...

Watch this Scientific Journal Video about A Rat Carotid Balloon Injury Model to Test Anti-vascular Remodeling Therapeutics at JoVE.com

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

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.

Rat Karotis Balon Yaralanma Modeli Anti-vasküler Tadilat Therapeutics 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 ...

The overall goal of this Rat Carotid Balloon Injury procedure is to mimic clinical angioplasty and its potential consequences. This message can help answer key questions in the cardiovascular research field, about the cardiovascular surgical injury elicited neointimal hyperplasia which can significantly compromise the benefits of the surgery. The main advantage of this technique is that the result can be obtained within two weeks, allowing a quick evaluation of the therapeutic reagents administered during the treatment period.Demonstrating the procedure will be Dr.Natalia Petrasheskatya from the laboratory of cardiovascular sciences and at the National Institute on Aging. Before beginning the surgery, autoclave all of the surgical instruments and use 70%ethanol to disinfect the operating platform surface. Put on the proper personal protective equipment, placing a box of sterilized gloves near the surgical platform for additional use.Next, fill a 1 milliliter syringe with sterile water and attach the syringe to a sterile, water-filled, two-way stopcock. Gently push the water through the stopcock to fill the lure-lock portion of the balloon catheter, and remove any air bubbled from the opening of the catheter. Then, test the balloon inflation to confirm that the balloon can be inflated with 0.02 milliliters of water.Now, check the pedal withdrawal reflex of an anesthetized 400 to 450 gram male Wistar rat. If the rat does not withdraw its foot, transfer the animal to the surgical platform. Cover the rat with a sterile surgical sheet, and cut the sheet to expose the neck region.Then adjust the animal into the supine position on a headed pad with the head toward the surgeon and tape the limbs into place. To isolate the left carotid artery, us a number 14 scalpel to make a straight incision from below the chin to the top of the sternum, just below the ribcage. Using a 7S forceps, blunt dissect under the skin and through the salivary gland tissues to expose the underlying muscle layer.Separate the muscle tissues to gain access to the carotid vasculature and the vagus nerve in the neck. Continue to blunt dissect the tissues surrounding the carotid artery, carefully separating the vagus nerve and the vascular fascia without damaging either tissue, until the bifurcation of the common carotid artery and the internal and external carotid branches are clearly visible. When the left common carotid artery has been exposed all the way to the sternum, place a 4-0 silk stay suture behind the artery as close to the sternum as possible.Permanently ligate the smaller arteries with the sutures, along the external carotid artery, including the ascending pharyngeal, the occipital, and the superior thyroid to avoid arterial leakage. Place another 4-0 silk stay suture immediately distal to the bifurcation, behind the external carotid artery as far from the bifurcation as possible. Then gently retract the overlying carotid artery to the right to visualize the internal carotid artery and place a 4-0 silk stay suture around the artery to avoid significant retrograde blood-loss.To introduce the balloon catheter, retract the proximal suture on the common carotid artery, and temporarily stop the blood flow with an arterial clamp. Make a small incision on the external carotid artery as distal to the suture as possible, and gently insert the uninflated balloon catheter into the incision. Advance the catheter into the lumen until it is close to the arterial clamp on the common carotid artery.Then remove the clamp and advance the catheter to the aortic arch, approximately 35 to 40 millimeters from the incision. Now use the syringe of sterile water to manually inflate the catheter to a 0.02 milliliter volume, and turn the stopcock to the locked position. Slowly withdraw the catheter while rotating to produce the injury.When the instrument is close to the arterial incision, deflate the balloon and retract the catheter back to its original position. After repeating the balloon injury procedure three times, carefully remove the catheter from the arterial lumen and close the arteriotomy with size 5-0 silk stay sutures. Release the internal carotid artery to restore the blood flow and check for arterial leakage.Finally, remove the stay sutures and clamps and close the glandular tissue with 6-0 silk sutures. Then, close the skin and monitor the animal until it is fully recovered. Two weeks after the balloon injury, the carotid arteries can be isolated for histomorphological analysis.The balloon injury elicits neointimal growth, or thickening of the vessel wall. If left untreated or treated with a saline control, compared to a non-operated vessel section from the same animal. By contrast, the vessel section from a rat that received a therapeutic reagent that blocks neointimal growth demonstrates a significantly reduced thickening in the vessel wall.The effects of the balloon injury and therapeutic treatment can also be evaluated in vivo, by ultrasound sonography. And these data collaborate well with the results from histomorphological analysis. When mastered, this techniques can be completed in 30 minutes, if it is performed properly.While attempting this procedure, it is important to remember to prepare and test the balloon catheter for leaks prior to the surgery. After watching this video, you should have a good understanding of how to perform the rat carotid balloon injury procedure, for evaluating tracks and therapeutic reagents that block neointimal growth and to study the mag-ni-sen of injury elicited neointimal growth. Don't forget that working with animals requires an approved protocol by your institutional AICUC.You should also consult with your institutional laboratory animal veterinarians to establish the best practices for post-surgical care, and monitoring of the animals.

Research

JoVE Journal

Medicine

Rat Model of Blood-brain Barrier Disruption to Allow Targeted Neurovascular Therapeutics

Kan-beyin bariyer bozulması arasında Rat Modeli Nörovasküler Therapeutics Hedefli izin ver için

Endothelial cells with tight junctions along with the basement membrane and astrocyte end feet surround cerebral blood vessels to form the blood-brain ...

Tıp

Carotid Artery Infusions for Pharmacokinetic and Pharmacodynamic Analysis of Taxanes in Mice

Farelerde Taksanlar Farmakokinetik ve Farmakodinamik Analiz Karotis Arter İnfüzyonlar

When proposing the use of a drug, drug combination, or drug delivery into a novel system, one must assess the pharmacokinetics of the drug in the study ...

Vascular Balloon Injury and Intraluminal Administration in Rat Carotid Artery

Sıçan Karotis Arter Vasküler Balon Yaralanma ve intraluminal İdaresi

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 ...

Retroductal Submandibular Gland Instillation and Localized Fractionated Irradiation in a Rat Model of Salivary Hypofunction

Tükürük hipofonksiyonunun bir Sıçan Modelinde Retroductal Submandibuler Bezi damlatma ve yerelleştirilmiş fraksiyonlara Işınlama

Normal tissues that lie within the portals of radiation are inadvertently damaged. Salivary glands are often injured during head and neck radiotherapy. ...

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

Tavşan modeli hızlandırılmış ateroskleroz: iliyak arter balon yaralanma bir metodolojik açıdan

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

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

Fare Abdominal aort Myointimal Hiperplazi ikna etmek için balon tabanlı yaralanma

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

A Drosophila In Vivo Injury Model for Studying Neuroregeneration in the Peripheral and Central Nervous System

A Drosophila In Vivo Injury Model for Studying Neuroregeneration in the Peripheral and Central Nervous System

Neuroregeneration periferik ve santral sinir sistemi çalışmak için bir Vivo Drosophila yaralanma modeli

The regrowth capacity of damaged neurons governs neuroregeneration and functional recovery after nervous system trauma. Over the past few decades, various ...

Ferric Chloride-induced Canine Carotid Artery Thrombosis: A Large Animal Model of Vascular Injury

Ferrik klorid kaynaklı köpek karotid arter trombozu: Bir büyük hayvan modeli vasküler yaralanma

Occlusive arterial thrombosis leading to cerebral ischemic stroke and myocardial infarction contributes to ~13 million deaths every year globally. Here, ...

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

Periadventitial Terapötik Uygulama ile Sıçan Karotis Arter Basınç Kontrollü Segmental Balon Yaralanması

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

A Rat Model of Pressure Overload Induced Moderate Remodeling and Systolic Dysfunction as Opposed to Overt Systolic Heart Failure

Aşırı Sistolik Kalp Yetmezliği nin Aksine Aşırı Basınç Ait Orta Remodeling ve Sistolik Disfonksiyon Bir Rat Modeli

In response to an injury, such as myocardial infarction, prolonged hypertension or a cardiotoxic agent, the heart initially adapts through the activation ...