Name: ferric chloride-induced thrombosis mouse model on carotid artery and mesentery vessel
Uploaded: 2015-06-29T14:00:00
Description: Watch this Scientific Journal Video about Ferric Chloride-induced Thrombosis Mouse Model on Carotid Artery and Mesentery Vessel at JoVE.com

Forfatterne vil gerne anerkende teknisk support fra Joy Yao og Dr. Karen Alt, samt finansiering fra NHMRC og NHF.

Alle forsøg med dyr blev godkendt af Alfred Medical Research og Uddannelse Precinct Animal Ethics Committee (E / 1534/2015 / B). Alle kirurgiske manipulationer blev udført i fuld bedøvelse og dyrene ikke oplever smerte på noget tidspunkt. Alle beskrevne forsøg er ikke-inddrivelse. 1. Fremstilling <ol><li> Skære tynde bånd af filterpapir (1 mm x 2 mm). </li><li> Frisk forberede 2 opløsninger af ferrichlorid på 4% (vægt / volumen) og 6% (vægt / volumen) fortyndet i deioniseret van...

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	<li>Leadley, R. J., Chi, L., Rebello, S. S., Gagnon, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Contribution+of+in+vivo+models+of+thrombosis+to+the+discovery+and+development+of+novel+antithrombotic+agents.">Contribution of in vivo models of thrombosis to the discovery and development of novel antithrombotic agents.</a> J Pharmacol Toxicol Methods. 43 (2), 101-116 (2000).</li><li>Johnson, G. J., Griggs, T. R., Badimon, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+utility+of+animal+models+in+the+preclinical+study+of+interventions+to+prevent+human+coronary+artery+restenosis:+analysis+and+recommendations.+On+behalf+of+the+Subcommittee+on+Animal,+Cellular+and+Molecular+Models+of+Thrombosis+and+Haemostasis+of+the+Scientific+and+Standardization+Committee+of+the+International+Society+on+Thrombosis+and+Haemostasis.">The utility of animal models in the preclinical study of interventions to prevent human coronary artery restenosis: analysis and recommendations. On behalf of the Subcommittee on Animal, Cellular and Molecular Models of Thrombosis and Haemostasis of the Scientific and Standardization Committee of the International Society on Thrombosis and Haemostasis.</a> Thromb Haemost. 81 (5), 835-843 (1999).</li><li>Day, S. M., Reeve, J. L., Myers, D. D., Fay, W. 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Ferrichlorid induceret thrombosemodel er et fremragende forskningsværktøj. Som vist i denne undersøgelse, er det ekstremt let at implementere og når de anvendes i kombination med intravital mikroskopi eller Doppler flowmeter, det giver en god overvågning i realtid af thrombedannelse. Justering af tid eksponering og koncentrationen af FeCI3, det giver også mulighed for at producere enten ikke-okklusive eller okklusive tromber. Men denne fremgangsmåde har også nogle begræns...

Studiet af de mekanismer, der er involveret i udviklingen af ​​trombose og evaluering af effektiviteten af ​​anti-trombotiske lægemidler kræver veletableret eksperimentelle dyremodeller. Store dyremodeller var de første til at blive brugt, da de giver store skibe mere ligner mennesker end gnavere 1. Men høje omkostninger, de større nødvendige faciliteter og vanskeligheden ved at manipulere dem er genetisk er store ulemper for deres brug og store dyr er nu begrænset til sene prækliniske studier, når foreløbige test på gnavere har givet afgørende resultater 2. Med bred tilgængelighed af transgene og knockout stammer og deres lille størrelse, der minimerer mængden af antitrombotiske lægemidler kræves for in vivo testning mus hovedsageligt bruges til trombose forskning. Derfor har flere modeller af trombotiske lidelser er udviklet i mus 3. Mange etablerede trombose modeller forstyrre Intimet lag af karvæggen, efterfulgt af udsættelse af det sub endotel ekstracellulære matrix til blodgennemstrømningen inducere dannelsen af blodpropper 4. Tromberne kan skyldes eksponering af kollagen, som udløser blodplader aktivering eller / og fra eksponering af vævsfaktor som aktiverer koagulationskaskaden 5. Adskillige teknikker anvendes derpå til at nå det oprindelige fartøj skade. Pierangeli et al. Udviklet en mekanisk sprængning model med en mikrokirurgi værktøj på den femorale vene 6. Kikushi et al. Beskrev en model, som består i administration af et billede reaktiv forbindelse (Rose Bengal), der akkumuleres i lipiddobbeltlaget af endotelceller, efterfulgt af det specifikke excitation af karvæggen af interesse med grønt lys (540 nm) 7. Skaden kan også induceres af en kort høj intensitet puls laserbelysning 8. En anden teknik for det første fastsat på halspulsåren hos rotterbestår i topisk påføring af ferrichlorid (FeCl3) 9. I dette tilfælde er de fartøj blotlægning resultater fra frie radikaler genereret af FeCl3 som forårsager lipidperoxidation og destruktion af endotelceller 10. Skaden inducerer ekspression af flere adhæsionsmolekyler udløser blodpladeadhæsion og aggregering såvel som leukocytter rekruttering. Det er blevet påvist, at leukocytter, især neutrofiler, spiller en afgørende rolle i aktiveringen af blodkoaguleringskaskaden fører til thrombose 11. Denne metode er velegnet til at reproducere koagulationskaskaden; efterforskere skal huske på, at i denne musemodel, er trombose typisk induceret i sunde fartøjer mens trombose hos mennesker primært forekommer i syge f.eks. atherosclerotiske fartøjer. Da denne model er meget enkel at implementere og er også effektiv i mus, er det nu det mest brugt trombose tilstandl for lille dyr in vivo studier. Desuden er denne teknik giver mulighed for at inducere dannelsen af ​​tromber i forskellige typer beholdere. Target fartøjer kan være arterier eller vener med stor diameter (carotis, femorale, vena cava) eller lille diameter (mesenterium, cremaster) 12-14. For nylig blev det også anvendt på den proksimale arteria cerebri media at udvikle en model for slagtilfælde 15. Den trombose dannelse kan være direkte observeret af intravital mikroskopi efter fluorescerende mærkning af blodplader og leukocytter eller overvåges ved at måle blodgennemstrømningen faldet med en temperaturføler eller en Doppler probe 12,16,17. Adskillige parametre såsom okklusion tid, thrombedannelse tid eller trombe størrelse kan derefter undersøges. De fysiologiske forskelle mellem fartøjer undersøgte resultat i betydelige variationer i tromber opnået. Derfor efterforskere vælge normalt målkarret i henhold til de parametre, som de ønsker at measure og / eller sygdommen indstilling de ønsker at undersøge. Typisk model på halspulsåren er mere relevant for forskning i aterotrombose relateret til myokardieinfarkt eller slagtilfælde mens undersøgelser af vena cava er mere relevante for forskning i dyb venetrombose. Tilgængeligheden af ​​de forskellige fartøjer bestemmer også den metode, der anvendes til at måle trombe vækst. For eksempel, er let at få adgang gør denne model velegnet til intravital mikroskopisk observation og undersøgelse af dynamikken i trombedannelse de mesenteriske fartøjer. Halspulsåren er mindre tilgængeligt men større muliggør blod flowmålinger og giver en fremragende model til at studere okklusiv thrombose. Ferrichlorid induceret thrombosemodel har tilvejebragt enorme fremskridt i forståelsen af ​​denne patologi. Det har været anvendt i mange undersøgelser, der fokuserer på den rolle, von Willebrand faktor i trombose dannelse 18,19. Kombineret med genetisk modifikation teknikker, har det kunnet pege på mange specifikke gen involveret i trombotiske lidelser. Lamrani et al. for eksempel har vist, at en knock-in for JAK2 V617F-genet er associeret med en accelererende dannelse af ustabile koagel 20. Zhang et al. Har undersøgt den fysiologiske konsekvenser af P2Y12 blodplade-receptoren og viste, at transgene mus, der overudtrykker specifikt denne receptor i blodplader kun, udviste en mere hurtig og stabil thrombedannelse i mesenterialarterie såret med FeCl3 21. Den afgørende rolle, Tissue-plasminogenaktivator (tPA) og urokinase-plasminogenaktivator (uPA) i fibrin nedbrydningsprocessen er også blevet undersøgt i denne fremgangsmåde 22. Endvidere denne model giver også en enkel og nøjagtig måde at teste de fibrinolytiske kapacitet i mange nye lægemidler in vivo. For eksempel har Wang et al. Brugt denne model til the præklinisk validering af en hidtil ukendt rekombinant plasminogenaktivator målrettet mod aktiverede blodplader 23. Denne metode også gjort det muligt validering af terapeutiske proteiner isoleret fra spyt af flåter, vampyr flagermus, og myg eller fra giften af slanger med specifik identifikation af målet 24-27. Disse eksempler viser alsidigheden af ​​den ferrichlorid model. I denne artikel har vi fokus på to metoder og studere ferrichlorid induceret trombose på to forskellige skibstype; mesenteriske fartøj og halspulsåren.

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			<td>Whatman chromatography paper</td>
			<td>GE Healthcare</td>
			<td>3030917</td>
			<td></td>
		</tr>
		<tr>
			<td>Iron (III) chloride 40 % (w/v)</td>
			<td>VWR</td>
			<td>24212.298</td>
			<td></td>
		</tr>
		<tr>
			<td>Rhodamine 6G</td>
			<td>Sigma</td>
			<td>R4127</td>
			<td></td>
		</tr>
		<tr>
			<td>Inverted microscope&#160;</td>
			<td>Olympus</td>
			<td>IX81</td>
			<td></td>
		</tr>
		<tr>
			<td>Digital black-and-white camera&#160;</td>
			<td>Olympus</td>
			<td>XM10</td>
			<td></td>
		</tr>
		<tr>
			<td>Doppler flowmeter</td>
			<td>Transonic</td>
			<td>TS420</td>
			<td></td>
		</tr>
		<tr>
			<td>Nano-doppler flow probe</td>
			<td>Transonic</td>
			<td>0.5 PBS</td>
			<td></td>
		</tr>
		<tr>
			<td>Ketamine</td>
			<td>Hospira&#160;</td>
			<td>0409-2051-05</td>
			<td></td>
		</tr>
		<tr>
			<td>Xylazine (Rampun)</td>
			<td>Bayer</td>
			<td>75313&#160;</td>
			<td></td>
		</tr>
		<tr>
			<td>Petri dish</td>
			<td>Sarstedt</td>
			<td>82.1472</td>
			<td></td>
		</tr>
		<tr>
			<td>Insulin syringe (29 G)</td>
			<td>BD Ultra-Fine</td>
			<td>326103</td>
			<td></td>
		</tr>
		<tr>
			<td>Cotton tipped applicators</td>
			<td>BSN medical</td>
			<td>211827A</td>
			<td></td>
		</tr>
		<tr>
			<td>Dynek dysilk sutures</td>
			<td>Dynek Pty Ltd</td>
			<td>CS30100</td>
			<td></td>
		</tr>
		<tr>
			<td>Dulbecco&#39;s phosphate buffer saline (PBS)</td>
			<td>Gibco life technologies</td>
			<td>21600-069</td>
			<td></td>
		</tr>
		<tr>
			<td>Heating pad</td>
			<td>Kirchner</td>
			<td>T60</td>
			<td></td>
		</tr>
	</tbody>
</table>

ferric chloride-induced thrombosis mouse model on carotid artery and mesentery vessel

Severe thrombosis and its ischemic consequences such as myocardial infarction, pulmonary embolism and stroke are major worldwide health issues. The ferric chloride injury is now a well-established technique to rapidly and accurately induce the formation of thrombi in exposed veins or artery of small and large diameter. This model has played a key role in the study of the pathophysiology of thrombosis, in the discovery and validation of novel antithrombotic drugs and in the understanding of the mechanism of action of these new agents. Here, the implementation of this technique on a mesenteric vessel and carotid artery in mice is presented. The method describes how to label circulating leukocytes and platelets with a fluorescent dye and to observe, by intravital microscopy on the exposed mesentery, their accumulation at the injured vessel wall which leads to the formation of a thrombus. On the carotid artery, the occlusion caused by the clot formation is measured by monitoring the blood flow with a Doppler probe.

Den fluorescerende intravital mikroskopi observation af mesenteriet vil afsløre akkumuleringen af rhodamin 6G mærkede blodplader og leukocytter langs karvæggen såret af FeCl3. Den gradvise dannelse af en delvis trombe overvåges i en 200 um mesenteriet fartøj (figur 1). En thrombe langsomt vises, og er klart identificerbare efter det første minut af eksponering for FeCl3 (figur 1, t = 60 sek). 40 sekunder efter fjernelse af filtrerpapiret gennemvædet med <su...

Watch this Scientific Journal Video about Ferric Chloride-induced Thrombosis Mouse Model on Carotid Artery and Mesentery Vessel at JoVE.com

Ferric Chloride-induced Thrombosis Mouse Model on Carotid Artery and Mesentery Vessel

The FeCl3 induced thrombosis model in mice is described herein. A method to monitor thrombus growth by intravital microscopy observation on a mesenteric vessel and by blood flow measurement in the carotid artery is presented.

Ferric Chloride-induceret Trombose Mouse Model på Arteria carotis og mesenterium Vessel

Severe thrombosis and its ischemic consequences such as myocardial infarction, pulmonary embolism and stroke are major worldwide health issues. The ferric ...

ferric-chloride-induced-thrombosis-mouse-model-on-carotid-artery

Research

JoVE Journal

Medicine

A Model of Disturbed Flow-Induced Atherosclerosis in Mouse Carotid Artery by Partial Ligation and a Simple Method of RNA Isolation from Carotid Endothelium

Despite the well-known close association, direct evidence linking disturbed flow to atherogenesis has been lacking. We have recently used a modified version of carotid partial ligation methods [1,2] to show that it acutely induces low and oscillatory flow conditions, two key characteristics of disturbed flow, in the mouse common carotid artery. Using this model, we have provided direct evidence that disturbed flow indeed leads to rapid and robust atherosclerosis development in Apolipoprotein E knockout mouse [3]. We also developed a method of endothelial RNA preparation with high purity from the mouse carotid intima [3]. Using this mouse model and method, we found that partial ligation causes endothelial dysfunction in a week, followed by robust and rapid atheroma formation in two weeks in a hyperlipidemic mouse model along with features of complex lesion formation such as intraplaque neovascularization by four weeks. This rapid in vivo model and the endothelial RNA preparation method could be used to determine molecular mechanisms underlying flow-dependent regulation of vascular biology and diseases. Also, it could be used to test various therapeutic interventions targeting endothelial dysfunction and atherosclerosis in considerably reduced study duration.

This describes a partial carotid ligation surgery, which causes disturbed flow conditions and subsequent atherosclerosis development (in two weeks) with intraplaque neo-vascularization (in four weeks) in the mouse common carotid artery. We also describe a novel method of RNA isolation from the carotid intima, providing high purity endothelial RNA.

Despite the well-known close association, direct evidence linking disturbed flow to atherogenesis has been lacking. We have recently used a modified ...

Mouse Complete Stasis Model of Inferior Vena Cava Thrombosis

Venous thromboembolism (VTE) includes both deep vein thrombosis (DVT) and pulmonary embolism (PE). In the United States (U.S.), the high morbidity and mortality rates make VTE a serious health concern 1-2. After heart disease and stroke, VTE is the third most common vascular disease 3. In the U.S. alone, there is an estimated 900,000 people affected each year, with 300,000 deaths occurring annually 3. A reliable in vivo animal model to study the mechanisms of this disease is necessary.
The advantages of using the mouse complete stasis model of inferior vena cava thrombosis are several. The mouse model allows for the administration of very small volumes of limited availability test agents, reducing costs dramatically. Most promising is the potential for mice with gene knockouts that allow specific inflammatory and coagulation factor functions to be delineated. Current molecular assays allow for the quantitation of vein wall, thrombus, whole blood, and plasma for assays. However, a major concern involving this model is the operative size constraints and the friability of the vessels. Also, due to the small IVC sample weight (mean 0.005 grams) it is necessary to increase animal numbers for accurate statistical analysis for tissue, thrombus, and blood assays such as real-time polymerase chain reaction (RT-PCR), western blot, enzyme-linked immunosorbent (ELISA), zymography, vein wall and thrombus cellular analysis, and whole blood and plasma assays 4-8.
The major disadvantage with the stasis model is that the lack of blood flow inhibits the maximal effect of administered systemic therapeutic agents on the thrombus and vein wall.

The mouse complete stasis model of inferior vena cava thrombosis yields quantifiable amounts of vein wall tissue and thrombus. It has proven useful for evaluating interactions between the vein wall and the occlusive thrombus and in assessing the progression from acute to chronic inflammation.

Venous thromboembolism (VTE) includes both deep vein thrombosis (DVT) and pulmonary embolism (PE).  In the United States (U.S.), the high morbidity and ...

Mouse Model of Middle Cerebral Artery Occlusion

Stroke is the most common fatal neurological disease in the United States 1. The majority of strokes (88%) result from blockage of blood vessels in the brain (ischemic stroke) 2. Since most ischemic strokes (~80%) occur in the territory of middle cerebral artery (MCA) 3, many animal stroke models that have been developed have focused on this artery. The intraluminal monofilament model of middle cerebral artery occlusion (MCAO) involves the insertion of a surgical filament into the external carotid artery and threading it forward into the internal carotid artery (ICA) until the tip occludes the origin of the MCA, resulting in a cessation of blood flow and subsequent brain infarction in the MCA territory 4. The technique can be used to model permanent or transient occlusion 5. If the suture is removed after a certain interval (30 min, 1 h, or 2 h), reperfusion is achieved (transient MCAO); if the filament is left in place (24 h) the procedure is suitable as a model of permanent MCAO. This technique does not require craniectomy, a neurosurgical procedure to remove a portion of skull, which may affect intracranial pressure and temperature 6. It has become the most frequently used method to mimic permanent and transient focal cerebral ischemia in rats and mice 7,8. To evaluate the extent of cerebral infarction, we stain brain slices with 2,3,5-triphenyltetrazolium chloride (TTC) to identify ischemic brain tissue 9. In this video, we demonstrate the MCAO method and the determination of infarct size by TTC staining.

We demonstrate in the video a method for producing a middle cerebral artery occlusion in adult mice using an intraluminal monofilament. We also show how to evaluate the extent of cerebral infarction by 2,3,5-triphenyltetrazolium chloride (TTC) staining.

Stroke is the most common fatal neurological disease in the United States 1. The majority of strokes (88%) result from blockage of blood vessels in the ...

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.

En musemodel af Stent Implantation i halspulsåren for Studiet af Restenosis

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

Adult Mouse Venous Hypertension Model: Common Carotid Artery to External Jugular Vein Anastomosis.

The understanding of the pathophysiology of brain arteriovenous malformations and arteriovenous fistulas has improved thanks to animal models. A rat model creating an artificial fistula between the common carotid artery (CCA) and the external jugular vein (EJV) has been widely described and proved technically feasible. This construct provokes a consistent cerebral venous hypertension (CVH), and therefore has helped studying the contribution of venous hypertension to formation, clinical symptoms, and prognosis of brain AVMs and dural AVFs. Equivalent mice models have been only scarcely described and have shown trouble with stenosis of the fistula. An established murine model would allow the study of not only pathophysiology but also potential genetic therapies for these cerebrovascular diseases.
We present a model of arteriovenous fistula that produces a durable intracranial venous hypertension in the mouse. Microsurgical anastomosis of the murine CCA and EJV can be difficult due to diminutive anatomy and frequently result in a non-patent fistula. In this step-by-step protocol we address all the important challenges encountered during this procedure. Avoiding excessive retraction of the vein during the exposure, using 11-0 sutures instead of 10-0, and making a carefully planned end-to-side anastomosis are some of the critical steps. Although this method requires advanced microsurgical skills and a longer learning curve that the equivalent in the rat, it can be consistently developed.
This novel model has been designed to integrate transgenic mouse techniques with a previously well-established experimental system that has proved useful to study brain AVMs and dural AVFs. By opening the possibility of using transgenic mice, a broader spectrum of valid models can be achieved and genetic treatments can also be tested. The experimental construct could also be further adapted to the study of other cerebrovascular diseases related with venous hypertension such as migraine, transient global amnesia, transient monocular blindness, etc.

We describe a method for creating a reliable model of cerebral venous hypertension in the adult mouse. This model has been widely described and tested in the rat. This new counterpart in the mice opens the possibility of using genetic modified animals and thereby broadens the applications of the model.

Voksen Mouse Venøs Hypertension Model: carotis communis til eksterne halsvene anastomose.

The understanding of the pathophysiology of brain arteriovenous malformations and arteriovenous fistulas has improved thanks to animal models. A rat model ...

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.

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 ...

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.

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 ...

Ferric Chloride-induced Murine Thrombosis Models

Arterial thrombosis (blood clot) is a common complication of many systemic diseases associated with chronic inflammation, including atherosclerosis, diabetes, obesity, cancer and chronic autoimmune rheumatologic disorders. Thrombi are the cause of most heart attacks, strokes and extremity loss, making thrombosis an extremely important public health problem. Since these thrombi stem from inappropriate platelet activation and subsequent coagulation, targeting these systems therapeutically has important clinical significance for developing safer treatments. Due to the complexities of the hemostatic system, in vitro experiments cannot replicate the blood-to-vessel wall interactions; therefore, in vivo studies are critical to understand pathological mechanisms of thrombus formation. To this end, various thrombosis models have been developed in mice. Among them, ferric chloride (FeCl3) induced vascular injury is a widely used model of occlusive thrombosis that reports platelet activation and aggregation in the context of an aseptic closed vascular system. This model is based on redox-induced endothelial cell injury, which is simple and sensitive to both anticoagulant and anti-platelets drugs. The time required for the development of a thrombus that occludes blood flow gives a quantitative measure of vascular injury, platelet activation and aggregation that is relevant to thrombotic diseases. We have significantly refined this FeCl3-induced vascular thrombosis model, which makes the data highly reproducible with minimal variation. Here we describe the model and present representative data from several experimental set-ups that demonstrate the utility of this model in thrombosis research.

We report a refined procedure of the ferric chloride (FeCl3)-induced thrombosis models on carotid and mesenteric artery as well as vein, characterized efficiently using intravital microscopy to monitor time to occlusive thrombi formation.

Ferrichlorid-induceret murin Trombose Modeller

Arterial thrombosis (blood clot) is a common complication of many systemic diseases associated with chronic inflammation, including atherosclerosis, ...

Phase Contrast Magnetic Resonance Imaging in the Rat Common Carotid Artery

Phase contrast magnetic resonance imaging (PC-MRI) is a noninvasive approach that can quantify flow-related parameters such as blood flow. Previous studies have shown that abnormal blood flow may be associated with systemic vascular risk. Thus, PC-MRI can facilitate the translation of data obtained from animal models of cardiovascular diseases to pertinent clinical investigations. In this report, we describe the procedure for measuring blood flow in the common carotid artery (CCA) of rats using cine-gated PC-MRI and discuss relevant analysis methods. This procedure can be performed in a live, anesthetized animal and does not require euthanasia after the procedure. The proposed scanning parameters yield repeatable measurements for blood flow, indicating excellent reproducibility of the results. The PC-MRI procedure described in this article can be used for pharmacological testing, pathophysiological assessment, and cerebral hemodynamics evaluation.

The overall goal of this procedure is to measure the blood flow in the rat common carotid artery by using noninvasive phase contrast magnetic resonance imaging.

Fase kontrast magnetisk resonans i rotte fælles halspulsåren

Phase contrast magnetic resonance imaging (PC-MRI) is a noninvasive approach that can quantify flow-related parameters such as blood flow. Previous ...

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

Occlusive arterial thrombosis leading to cerebral ischemic stroke and myocardial infarction contributes to ~13 million deaths every year globally. Here, we have translated a vascular injury model from a small animal into a large animal (canine), with slight modifications that can be used for pre-clinical screening of prophylactic and thrombolytic agents. In addition to the surgical methods, the modified protocol describes the step-by-step methods to assess carotid artery canalization by angiography, detailed instructions to process both the brain and carotid artery for histological analysis to verify carotid canalization and cerebral hemorrhage, and specific parameters to complete an assessment of downstream thromboembolic events by utilizing magnetic resonance imaging (MRI). In addition, specific procedural changes from the previously well-established small animal model necessary to translate into a large animal (canine) vascular injury are discussed.

Here, we present the modifications necessary to a well characterized and commonly used small animal ferric chloride-induced (FeCl3) carotid artery injury model for use in a large animal vascular injury model. The resulting model can be utilized for pre-clinical trial assessment of both prophylactic and thrombolytic pharmacological and mechanical interventions.

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, ...