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.

BEMÆRK: brugen af ​​rotte ballon skade model og tilhørende procedurer, herunder injektion af rekombinante sRAGE og ultralyd sonografiske undersøgelser er blevet godkendt af Animal Care og brug Udvalg (ACUC) af National Institute on Aging, NIH. 1. Pre-kirurgi Forberedelser <ol><li> Instrument, Kirurgisk Platform, og personlige værnemidler. <ol><li> Se Materialer og udstyr Table for alle de kirurgiske instrumenter og reagenser, der anvendes i denne p...

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	<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>

Der har været to metoder, der anvendes til at puste ballonen op for at generere den skade, der fjerner maleriet endotel i de carotide arterielle lumen. Den ene er at fylde sprøjten med væske 20, og den anden er at benytte lufttryk 21. Vi foretrækker at bruge flydende fyldt injektionssprøjte fordi det nøjagtige væskevolumen (0,02 ml) vil blive anvendt til hver procedure. Dette gør en præcis og reproducerbar oppustning af ballonen, hvilket fører til et tilsvarende niveau for skade i hvert d...

Angioplasti er en endovaskulær procedure, der anvendes til at udvide forsnævrede eller spærret arterier resulterende fra patologiske tilstande, såsom aterosklerose. En almindelig komplikation ved angioplasti er den post-operative neointimal hyperplasi, eller restenose, som opstår på grund af kirurgiske skader og den efterfølgende inflammation-induceret vaskulær remodellering. Disse betingelser fører til proliferation af vaskulære glatte celler, og flere patofysiologiske konsekvenser 1-3. Neointimal hyperplasi re-fortykker fartøjet, og forekommer hos op til 60% post-angioplastik patienter inden for det første år. Derfor restenose er et alvorligt tilbageslag for udbredte angioplastik procedure 4. Selv om implantation af den lægemiddel-eluering stent eventuelt forebygge restenose, kan kun udvalgte kandidater gennemgå denne bekostelig procedure 5. Både dyr og kliniske undersøgelser har fastslået, at kronisk inflammation genereres af vascular skade og / eller kirurgisk sår fungerer som det vigtigste incitament for post-angioplastik neointimal vækst 2,4. De rotte carotis ballon skade model efterligner den kliniske situation og derfor fungerer som et værdifuldt modelsystem til at identificere cellulære faktorer, der involverer i den vaskulære remodellering og vaskulær proliferation 6-9. Denne model system er også et meget nyttigt redskab til at vurdere og / eller skærm for narkotika og terapeutiske reagenser, der undertrykker neointimal vækst i prækliniske translationelle studier 10-14. Sammenlignet med den murine carotid wire skade model 15 og den murine femorale arterie wire skade model 16, rotte carotid ballonskade model har fordelene ved at være tilstrækkelig stor i størrelse for at lette kirurgisk procedure, der letter reproducerbarheden af skade. Det kan give et større antal primære celler (fx vaskulære glatte muskelceller, endotelceller) for additional in vitro-undersøgelser for at afgrænse de molekylære mekanisme regulerer vaskulær remodellering. Vigtigt er, sammenlignet med mus, rotter vides også at være en bedre model for fysiologiske og toksikologiske undersøgelser 17. Selv om en ulempe eller begrænsning af rotte-modellen er manglen af ​​genetiske modificerede og gen-knockout-modeller, kan denne ulempe overvindes ved tilgængeligheden af ​​rotte genomiske sekvens og den seneste udvikling af kraftfulde genomiske redigeringsværktøjer såsom CRISPR-CAS-teknologi, der gør mulig manipulation af store serier af genomiske sekvenser i forskellige modelsystemer 18,19. Selv rotte ballon skade model er blevet anvendt af flere laboratorier og forskellige omfattende protokoller er blevet offentliggjort 20,21, denne protokol sigter på at give flere detaljer på forhånd kirurgi præparater og kan vejlede forskerne nye til denne procedure for at oprette denne kirurgiske praksis. Vi understreger også den post-kirurgi pleje af than dyr, der tillader ikke kun post mortem patologiske og histomorfologisk analyser af de terapeutiske virkninger på arterielle remodeling, men også ultralyd sonografiske undersøgelser i levende dyr 13,22.

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			<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>
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			<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>
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			<td height="21" style="height:21px;width:287px;">Self-regulating heating pad</td>
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			<td height="21" style="height:21px;width:287px;">ADS200 anesthesia system/ventilator</td>
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			<td height="21" style="height:21px;width:287px;">Isoflurane (forane), liquid form</td>
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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.

To uger efter ballonskade, er rotten aflivet, og carotidarterierne isoleres for histo-morfologisk analyse. Begge betjente venstre halspulsåre og un-betjente højre arterie er tværsnit, behandles og paraffin-indstøbt. Paraffinen Prøverne derefter yderligere tynd-snittet og farvet med hematoxylin-eosin (H &amp; E). Histo-morfologiske analyser udføres under anvendelse af en digital billedbehandling analysesystem. Detaljerne i arterie høst og histo-morfologisk analyse er blevet beskrev...

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.

En Rat carotis ballon Skade Model til Test Anti-vaskulære 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 ...

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

Rottemodel af blod-hjerne barrieren Disruption at muliggøre målrettede neurovaskulær Therapeutics

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

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

Halspulsåren infusioner farmakokinetisk og farmakodynamisk analyse af Taxaner i mus

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

Vascular Balloon Skade og intraluminal Administration i Rat Arteria carotis

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

Retroductal Submandibularis Gland Instillation og Lokaliseret Fraktioneret Bestråling i en rotte model af spyt hypofunktion

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

Kanin modellen af accelereret åreforkalkning: en metodologisk perspektiv iliaca arterie ballon skade

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

Ballon-baserede skade til at fremkalde Myointimal hyperplasi i mus Abdominal Aorta

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

En Drosophila In Vivo skade Model til at studere Neuroregeneration i perifere og centrale nervesystem

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

Ferrichloridopløsning-induceret Canine carotis arterie trombose: En stor dyr Model af vaskulære skade

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

En rotte halspulsåren Tryk-kontrolleret segmental ballon skade med Periadventitial Terapeutisk Application

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

En rotte model af trykoverbelastning induceret moderat remodellering og systolisk dysfunktion i modsætning til Åbenlys systolisk hjertesvigt

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