We are grateful to Dr. Clowes for first developing and describing this method. We are also thankful to Dr. Tulis for his detailed protocol which has been fundamentally helpful to our previous, current and future work. This work was supported by grants R01HL097111 and R01HL123364 from the NIH to M.T., and by American Heart Association grant 14GRNT18880008 to M.T.
We would like to thank Rachel Newton for her expert technical support and for her valuable help during the filming process.

NOTE: The use of animals for the following experiments has been reviewed and approved by the Institutional Animal Care and Use Committee (IACUC).
1. Preoperative Procedures
<ol style="margin-left: 40px;">
	<li>Sterilize surgical instruments before use.
	<ol>
		<li>Autoclave all surgical instruments 24 hr or less before surgery. If multiple surgeries are performed on the same day, sterilize the instruments by a dry bead sterilizer in between surgeries.</li>
	</ol>
	</li>
	<li>Filter-sterilize the saline solution ...

<ol>
	<li>Landzberg, B. R., Frishman, W. H., Lerrick, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pathophysiology+and+pharmacological+approaches+for+prevention+of+coronary+artery+restenosis+following+coronary+artery+balloon+angioplasty+and+related+procedures.">Pathophysiology and pharmacological approaches for prevention of coronary artery restenosis following coronary artery balloon angioplasty and related procedures.</a> Progress in Cardiovascular Diseases. 39, 361-398 (1997).</li><li>Muller, D. W., Ellis, S. G., Topol, E. 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+models+of+coronary+artery+restenosis.">Experimental models of coronary artery restenosis.</a> J. Am. Coll. Cardiol. 19, 418-432 (1992).</li><li>Clowes, A. W., Reidy, M. A., Clowes, M. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Kinetics+of+cellular+proliferation+after+arterial+injury.+I.+Smooth+muscle+growth+in+the+absence+of+endothelium.">Kinetics of cellular proliferation after arterial injury. I. Smooth muscle growth in the absence of endothelium.</a> Laboratory Investigation: A Journal of Technical Methods and Pathology. 49, 327-333 (1983).</li><li>Clowes, A. W., Reidy, M. A., Clowes, M. M. <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+stenosis+after+arterial+injury.">Mechanisms of stenosis after arterial injury.</a> Laboratory Investigation: A Journal of Technical Methods and Pathology. 49, 208-215 (1983).</li><li>Clowes, A. W., Clowes, M. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Kinetics+of+cellular+proliferation+after+arterial+injury.+IV.+Heparin+inhibits+rat+smooth+muscle+mitogenesis+and+migration.">Kinetics of cellular proliferation after arterial injury. IV. Heparin inhibits rat smooth muscle mitogenesis and migration.</a> Circulation Research. 58, 839-845 (1986).</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, 1362-1365 (2000).</li><li>Kiechl, S., Willeit, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+natural+course+of+atherosclerosis.+Part+II:+vascular+remodeling.">The natural course of atherosclerosis. Part II: vascular remodeling.</a> Bruneck Study Group. Arteriosclerosis, Thrombosis, and Vascular Biology. 19, 1491-1498 (1999).</li><li>Yamamoto, S., 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=Derivation+of+rat+embryonic+stem+cells+and+generation+of+protease-activated+receptor-2+knockout+rats.">Derivation of rat embryonic stem cells and generation of protease-activated receptor-2 knockout rats.</a> Transgenic Research. 21, 743-755 (2012).</li><li>Intengan, H. D., Schiffrin, E. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Vascular+remodeling+in+hypertension:+roles+of+apoptosis,+inflammation,+and+fibrosis.">Vascular remodeling in hypertension: roles of apoptosis, inflammation, and fibrosis.</a> Hypertension. 38, 581-587 (2001).</li><li>Meng, 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=Complex+hemodynamics+at+the+apex+of+an+arterial+bifurcation+induces+vascular+remodeling+resembling+cerebral+aneurysm+initiation.">Complex hemodynamics at the apex of an arterial bifurcation induces vascular remodeling resembling cerebral aneurysm initiation.</a> Stroke. 38, 1924-1931 (2007).</li><li>Sun, C. K., Shao, P. L., Wang, C. J., Yip, H. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Study+of+vascular+injuries+using+endothelial+denudation+model+and+the+therapeutic+application+of+shock+wave:+a+review.">Study of vascular injuries using endothelial denudation model and the therapeutic application of shock wave: a review.</a> American Journal of Rranslational Research. 3, 259-268 (2011).</li><li>Zhang, W., 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=Orai1-mediated+I+(CRAC)+is+essential+for+neointima+formation+after+vascular+injury.">Orai1-mediated I (CRAC) is essential for neointima formation after vascular injury.</a> Circulation Research. 109, 534-542 (2011).</li><li>Ollinger, 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=Bilirubin:+a+natural+inhibitor+of+vascular+smooth+muscle+cell+proliferation.">Bilirubin: a natural inhibitor of vascular smooth muscle cell proliferation.</a> Circulation. 112, 1030-1039 (2005).</li><li>Levitzki, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=PDGF+receptor+kinase+inhibitors+for+the+treatment+of+restenosis.">PDGF receptor kinase inhibitors for the treatment of restenosis.</a> Cardiovascular Research. 65, 581-586 (2005).</li><li>Asahara, 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=Local+delivery+of+vascular+endothelial+growth+factor+accelerates+reendothelialization+and+attenuates+intimal+hyperplasia+in+balloon-injured+rat+carotid+artery.">Local delivery of vascular endothelial growth factor accelerates reendothelialization and attenuates intimal hyperplasia in balloon-injured rat carotid artery.</a> Circulation. 91, 2793-2801 (1995).</li><li>Lee, K. 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=Alpha-lipoic+acid+inhibits+fractalkine+expression+and+prevents+neointimal+hyperplasia+after+balloon+injury+in+rat+carotid+artery.">Alpha-lipoic acid inhibits fractalkine expression and prevents neointimal hyperplasia after balloon injury in rat carotid artery.</a> Atherosclerosis. 189, 106-114 (2006).</li><li>Ji, 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=MicroRNA+expression+signature+and+antisense-mediated+depletion+reveal+an+essential+role+of+MicroRNA+in+vascular+neointimal+lesion+formation.">MicroRNA expression signature and antisense-mediated depletion reveal an essential role of MicroRNA in vascular neointimal lesion formation.</a> Circulation Research. 100, 1579-1588 (2007).</li><li>Merlet, 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=miR-424/322+regulates+vascular+smooth+muscle+cell+phenotype+and+neointimal+formation+in+the+rat.">miR-424/322 regulates vascular smooth muscle cell phenotype and neointimal formation in the rat.</a> Cardiovascular Research. 98, 458-468 (2013).</li><li>Huang, J., Niu, X. L., Pippen, A. M., Annex, B. H., Kontos, C. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Adenovirus-mediated+intraarterial+delivery+of+PTEN+inhibits+neointimal+hyperplasia.">Adenovirus-mediated intraarterial delivery of PTEN inhibits neointimal hyperplasia.</a> Arteriosclerosis, Thrombosis, And Vascular Biology. 25, 354-358 (2005).</li><li>Gonzalez-Cobos, J. C., 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=Store-independent+Orai1/3+channels+activated+by+intracrine+leukotriene+C4:+role+in+neointimal+hyperplasia.">Store-independent Orai1/3 channels activated by intracrine leukotriene C4: role in neointimal hyperplasia.</a> Circulation Research. 112, 1013-1025 (2013).</li><li>Guzman, L. A., 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=Local+intraluminal+infusion+of+biodegradable+polymeric+nanoparticles.+A+novel+approach+for+prolonged+drug+delivery+after+balloon+angioplasty.">Local intraluminal infusion of biodegradable polymeric nanoparticles. A novel approach for prolonged drug delivery after balloon angioplasty.</a> Circulation. 94, 1441-1448 (1996).</li><li>Lipke, E. A., West, J. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Localized+delivery+of+nitric+oxide+from+hydrogels+inhibits+neointima+formation+in+a+rat+carotid+balloon+injury+model.">Localized delivery of nitric oxide from hydrogels inhibits neointima formation in a rat carotid balloon injury model.</a> Acta Biomaterialia. 1, 597-606 (2005).</li><li>Osterrieder, W., 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=Role+of+angiotensin+II+in+injury-induced+neointima+formation+in+rats.">Role of angiotensin II in injury-induced neointima formation in rats.</a> Hypertension. 18, II60-II64 (1991).</li><li>Powell, J. S., 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=Inhibitors+of+angiotensin-converting+enzyme+prevent+myointimal+proliferation+after+vascular+injury.">Inhibitors of angiotensin-converting enzyme prevent myointimal proliferation after vascular injury.</a> Science. 245, 186-188 (1989).</li><li>. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Does+the+new+angiotensin+converting+enzyme+inhibitor+cilazapril+prevent+restenosis+after+percutaneous+transluminal+coronary+angioplasty?+Results+of+the+MERCATOR+study:+a+multicenter,+randomized,+double-blind+placebo-controlled+trial.+Multicenter+European+Research+Trial+with+Cilazapril+after+Angioplasty+to+Prevent+Transluminal+Coronary+Obstruction+and+Restenosis+(MERCATOR)+Study+Group.">Does the new angiotensin converting enzyme inhibitor cilazapril prevent restenosis after percutaneous transluminal coronary angioplasty? 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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=Knockout+rats+via+embryo+microinjection+of+zinc-finger+nucleases.">Knockout rats via embryo microinjection of zinc-finger nucleases.</a> Science. 325, 433 (2009).</li><li>Tesson, 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=Knockout+rats+generated+by+embryo+microinjection+of+TALENs.">Knockout rats generated by embryo microinjection of TALENs.</a> Nature Biotechnology. 29, 695-696 (2011).</li><li>Li, 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=Heritable+gene+targeting+in+the+mouse+and+rat+using+a+CRISPR-Cas+system.">Heritable gene targeting in the mouse and rat using a CRISPR-Cas system.</a> Nature Biotechnology. 31, 681-683 (2013).</li><li>Potier, 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=Evidence+for+STIM1-+and+Orai1-dependent+store-operated+calcium+influx+through+ICRAC+in+vascular+smooth+muscle+cells:+role+in+proliferation+and+migration.">Evidence for STIM1- and Orai1-dependent store-operated calcium influx through ICRAC in vascular smooth muscle cells: role in proliferation and migration.</a> FASEB Journal : Official Publication Of The Federation Of American Societies For Experimental Biology. 23, 2425-2437 (2009).</li><li>Aubart, F. C., 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=RNA+interference+targeting+STIM1+suppresses+vascular+smooth+muscle+cell+proliferation+and+neointima+formation+in+the+rat.+Molecular+Therapy.">RNA interference targeting STIM1 suppresses vascular smooth muscle cell proliferation and neointima formation in the rat. Molecular Therapy.</a> The Journal Of The American Society Of Gene Therapy. 17, 455-462 (2009).</li><li>Berra-Romani, R., Mazzocco-Spezzia, A., Pulina, M. V., Golovina, V. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ca2++handling+is+altered+when+arterial+myocytes+progress+from+a+contractile+to+a+proliferative+phenotype+in+culture.+American+journal+of+physiology.">Ca2+ handling is altered when arterial myocytes progress from a contractile to a proliferative phenotype in culture. American journal of physiology.</a> Cell Physiology. 295, C779-C790 (2008).</li><li>Bisaillon, 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=Essential+role+for+STIM1/Orai1-mediated+calcium+influx+in+PDGF-induced+smooth+muscle+migration.+American+journal+of+physiology.">Essential role for STIM1/Orai1-mediated calcium influx in PDGF-induced smooth muscle migration. American journal of physiology.</a> Cell Physiology. 298, C993-C1005 (2010).</li><li>Tulis, D. 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The rat carotid balloon injury has been well described by Tulis in 200734. It has been comprehensively discussed all the details of this procedure by Dr. Tulis. The readers who are interested in performing this procedure are highly recommended to read Tulis&#8217; protocol. However, there is one thing we are not agreeing with Dr. Tulis: Instead of inflating the balloon with saline or any kind of liquid, we suggested to inflate it with air. According to our personal experience, inflating with liquid can hardly avoid air bubbles. In...

Balloon catheters are medical devices used in the procedure of angioplasty, for the purpose of widening obstructed site(s) of atheroma or thrombus in a blood vessel. The narrowed vessel lumen is forced to open up by the inflated balloon and blood supply would be restored sequentially to relieve downstream ischemia symptoms, such as angina, myocardial infarction, and leg pain. Nevertheless, the great success of angioplasty has been diminished by post-operative complications such as results from force causing vascular barotrauma (balloon injury), namely vessel wall remodeling and in many cases re-narrowing of the vessel lumen (restenosis)1.
A number of animal models have been developed mimicking the angioplasty procedure to help investigators understand mechanisms underlying the balloon-injury-related vessel wall remodeling2. Among all the animal species utilized for modeling, rat is the most frequently used one. Compared to rabbits, dogs and swine, the advantages of rats are their low cost, their relative ease of use and the current knowledge of rat physiology. Although mice have an added advantage in a wide range of genetically manipulated strains, the mice vessel is too small to insert a balloon catheter. Over the past three decades, experimental rats have allowed researchers to gain better understanding of the molecular and cellular mechanisms underpinning neointima formation and vascular remodeling3-6. Beyond balloon injury, vascular remodeling are also involved in most major vascular diseases, such as atherosclerosis7,8, hypertension9, and aneurysm10. Thus, knowledge gained through the balloon injury model is in general beneficial to overall vascular wall disease studies.
The overall goal of the rat balloon injury model is not only to further understand vascular diseases but also to test the potency of novel agents for disease control11,12. Current clinical drug treatment to restenosis is applied by drug-eluting stents placed via the vessel lumen right after angioplasty. In animal models, an efficient yet more economical way for new agent testing is a well-developed local intraluminal perfusion method. Candidate agents that have been tested through this method include small molecule drugs13,14, cytokine or growth factors15,16, gene manipulating agents (cDNA clones, siRNA, etc.)17-20, and novel pharmaceutical formulations21,22.
So far, the rat balloon injury model remains one of the most useful models for studying vascular diseases/disorders. It is the fundamental step from bench to bedside, usually as the first step moving from in vitro to in vivo, but it should not be the last one. The outcome of rat experiments needs to be deliberated and further characterized before translation into human clinical use, due to the difference in vascular beds and vessel anatomy as well as the intrinsic species differences between human and rat23-26. Nevertheless, it is still an essential tool in translational medical research. While such research used to be limited by the lack of genetically modified rats, it has no longer been an issue since novel genomic approaches such as zinc-finger nucleases27, TALENs28 and CRISPR-Cas29 have made knockout rats easily accessible.

<table border="0" cellpadding="0" cellspacing="0" width="1059">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:384px;">Name of Material/ Equipment</td>
			<td style="width:389px;">Company</td>
			<td style="width:119px;">Catalog Number</td>
			<td style="width:167px;">Comments/Description</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Fogarty balloon embolectomy catheters, 2 French&#160;</td>
			<td>Edwards Lifesciences, Germany</td>
			<td>&#160;120602F</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Deltaphase Operating Board - Includes 2 Pads &#38; 2 Insulators</td>
			<td>Braintree Scientific, Inc.</td>
			<td>39OP</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">&#160;LED light source</td>
			<td>Fisher Scientific</td>
			<td>12-563-501&#160;</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Hartmann Mosquito Forceps 4&#8221; curved</td>
			<td>Apiary Medical, Inc. San Diego, CA</td>
			<td>gS 22.1670</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Crile Retractor 4&#8221; double ended</td>
			<td>Apiary Medical, Inc.</td>
			<td>gS 34.1934</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Other surgical instruments</td>
			<td>Roboz Surgical Instrument Company, Inc., Gaithersburg, MD</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Peripheral Intravenous (I.V.) Cannula, 24G</td>
			<td>BD</td>
			<td>381312</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Ketamine HCl, 100mg/mL, 10mL</td>
			<td>Ketaset- Patterson Vet</td>
			<td>07-803-6637&#160;</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Xylazine (AnaSed),20mg/mL,20mL</td>
			<td>Ketaset- Patterson Vet</td>
			<td>07-808-1947</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Buprenex, 0.3mg/1ml (5 Ampules/Box)</td>
			<td>Ketaset- Patterson Vet</td>
			<td>07-850-2280</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Nair Baby Oil Hair Removal Lotion-9 oz</td>
			<td>Amazon/Walmart/CVS</td>
			<td>N/A</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Inflation Device</td>
			<td>Demax Medical</td>
			<td>DID30</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">D300 3-way Stopcock</td>
			<td>B.Braun Medical Inc.</td>
			<td>4599543</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Artificial Tears Ointment&#160;</td>
			<td>Rugby Laboratories, Duluth, GA</td>
			<td>N/A</td>
			<td></td>
		</tr>
	</tbody>
</table>

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.

Two weeks after injury, carotid arteries are harvested, sectioned and subjected to morphological analysis. Arteries are cross-sectioned and stained with H&#38;E (Figures 1, 2B, C and 3). Rat carotid artery wall contains four layers of elastic lamina, which appear as pink lines. The area between the outermost lamina, external elastic lamina (EEL) and the innermost lamina, internal elastic lamina (IEL) is the media smooth muscle layer (Figure 1). The area inside of the IEL is the intima, a...

Watch this Scientific Journal Video about Vascular Balloon Injury and Intraluminal Administration in Rat Carotid Artery at JoVE.com

Vascular Balloon Injury and Intraluminal Administration in Rat Carotid 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.

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 ...

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Research

JoVE Journal

Medicine

16.2K vistas.  State University of New York College of Nanoscale Science and Engineering (SUNY CNSE).  Este protocolo utiliza un catéter de balón para causar una lesión intraluminal de la arteria carótida de rata y en adelante provocar hiperplasia neointimal. Este es un modelo bien establecido para el estudio de los mecanismos de remodelación vascular en respuesta a una lesión. También es ampliamente utilizado para determinar la validez de los enfoques terapéuticos potenciales. 