1. Topo Anestesia Procedure <ol><li> Gli animali vengono rimossi dal loro gabbia e messi in anestesia pre-operatoria camera di induzione per l&#39;induzione di gas al 5% isoflurano e il 100% di ossigeno, ad una portata di 0,5 litri al minuto (vaporizzatore controllato). </li><li> I topi sono sedato nella camera di induzione dell&#39;anestesia, rimosso dalla camera e l&#39;addome ventrale è rasata con cesoie elettriche, o mantenuti all&#39;interno della camera se un depilatorio capelli è usato (1 minuto). </li><li> Mentre ancora sotto sedazione, sono pesati, gli occhi sono lubrificati con pomata oftalmica sterile e quindi posto in un recumbancy dorsale su un acqua calda in circolazione dispositivo di riscaldamento in anestesia generale, con un musetto, con isoflurano e ossigeno. A questo punto il tasso di ossigeno si riduce a 0,2 litri al minuto. </li><li> L&#39;addome viene spruzzato con una soluzione di clorexidina (clorexidina diacetato è rapidamente bacterialcidal e persistente) e si asciugò con garza sterile o tre volte fino a quando il sito chirurgico è pulito. </li></ol> 2. Micro Mouse-chirurgici e procedure di ripristino <ol><li> Una incisione della linea mediana ventrale (2cm) è fatto con le forbici iris attraverso la pelle e la parete addominale esporre l&#39;addome. Una garza sterile imbevuta 2x2in soluzione salina viene utilizzato per riflettere l&#39;intestino alla sinistra dell&#39;animale. Un dispositivo di immobilizzazione mouse è messo in atto a consentire la visualizzazione della vena cava inferiore (IVC). </li><li> A questo punto, un livello di manutenzione del 2% isoflurance e tasso di ossigeno al 100% di 0,2 litri / min viene utilizzato per il resto della procedura chirurgica. </li><li> Dissezione Blunt è facilitato utilizzando un tampone sterile applicatore ed extra delicato tessuto dell&#39;iride pinze mezzo curvo. Bisogna fare attenzione nel maneggiare il mouse per tessuti sono estremamente fragili. Una volta entrati nella cavità addominale, utilizzare una garza sterile imbevuta di soluzione fisiologica 4x4in riflettere e esteriorizzare l&#39;intestino su un telino sterile, permettendo così la visualizzazione della IVC. </li><li> Tutte le filiali indietro IVC, dalle vene renali alla biforcazione iliaca, mentre sono cauterizzato rami laterali sono legatura con 7-0 punti di sutura non reattivi prolene. </li><li> L&#39;IVC è separato dall&#39;aorta, appena inferiore alle vene renali. 7-0 prolene viene usato per legare la IVC. </li><li> Il sito laparotomia è chiuso in due strati di moda. Il nostro laboratorio utilizza Vicryl 5-0, una sutura sintetica, in un modello continuo per chiudere la parete addominale. Colla pelle adesivo utilizzato per chiudere la pelle. Non ci sono suture esterne per la rimozione. </li><li> I topi sono poi recuperati in una gabbia di topo individuale, ha osservato post-operatoria (45 minuti a 2 ore) sotto una lampada riscaldante (distanza minima di-24 cm di distanza dalla gabbia), poi tornati alle loro abitazioni originale. Farmaci antidolorifici e antinfiammatori può essere utilizzato se non interferisce con l&#39;obiettivo di studio. Giustificazione scientifica per trattenere farmaci per il dolore, veterinario, e l&#39;approvazione ICUCA sono necessari. Il nostro laboratorio ha scoperto che la buprenorfina, 0,05-0,1 mg / kg SQ è un analgesico ottimo pre-e post-operatoria per i topi. Inoltre, a 4mg/kg lidocaina (0,4 ml / kg di una soluzione all&#39;1%) può essere infusa nel sito di incisione, quando gli analgesici sistemici possono confondere i risultati sperimentali. </li><li> Sham animali di ogni gruppo sperimentale sottoposto la dissezione sopra utilizzate per l&#39;approccio chirurgico, ad eccezione del IVC o rami di scarico non sono legatura o cauterizzato. </li><li> L&#39;eutanasia è eseguita in conformità con le raccomandazioni stabilite dalla American Veterinary Medical Association Linee guida sull&#39;eutanasia per i roditori. </li></ol>

<ol>
	<li>Naess, I. A., Christiansen, S. C., Romundstad, P., Cannegieter, S. C., Rosendaal, F. R., Hammerstrom, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Incidence+and+mortality+of+venous+thrombosis:+a+population-based+study.">Incidence and mortality of venous thrombosis: a population-based study.</a> J Thromb Haemost. 5, 692-699 (2007).</li><li>White, R. H., Keenan, C. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+race+and+ethnicity+on+the+incidence+of+venous+thromboembolism.">Effects of race and ethnicity on the incidence of venous thromboembolism.</a> Thromb Res. 123, S11-S17 (2009).</li><li>Heit, 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=The+epidemiology+of+venous+thromboembolism+in+the+community.">The epidemiology of venous thromboembolism in the community.</a> Arterioscler Thromb Vasc Biol. 28, 370-372 (2008).</li><li>Henke, P. K., Varga, A., De, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Deep+vein+thrombosis+resolution+is+modulated+by+monocyte+CXCR2-mediated+activity+in+a+mouse+model.">Deep vein thrombosis resolution is modulated by monocyte CXCR2-mediated activity in a mouse model.</a> Arterioscler Thromb Vasc Biol. 24, 1130-1137 (2004).</li><li>McDonald, A. P., Meier, T. R., Hawley, A. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Aging+is+associated+with+impaired+thrombus+resolution+in+a+mouse+model+of+stasis+induced+thrombosis.">Aging is associated with impaired thrombus resolution in a mouse model of stasis induced thrombosis.</a> Thromb Res. 125, 72-78 (2010).</li><li>Myers, D., Farris, D., Hawley, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Selectins+influence+thrombosis+in+a+mouse+model+of+experimental+deep+venous+thrombosis.">Selectins influence thrombosis in a mouse model of experimental deep venous thrombosis.</a> J Surg Res. 108, 212-221 (2002).</li><li>Myers, D. D., Hawley, A. E., Farris, D. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=P-selectin+and+leukocyte+microparticles+are+associated+with+venous+thrombogenesis.">P-selectin and leukocyte microparticles are associated with venous thrombogenesis.</a> J Vasc Surg. 38, 1075-1089 (2003).</li><li>Myers, D. D. J. r., Rectenwald, J. E., Bedard, P. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Decreased+venous+thrombosis+with+an+oral+inhibitor+of+P+selectin.">Decreased venous thrombosis with an oral inhibitor of P selectin.</a> J Vasc Surg. 42, 329-336 (2005).</li></ol>

Nessun conflitto di interessi dichiarati.

Completa il modello di mouse Stasi di trombosi vena cava inferiore può essere eseguita su più ceppi di topi di diversa età. Il modello è più adatto ai topi di almeno 20 grammi di peso. Questo modello animale di trombosi venosa permette ai ricercatori di studiare le interazioni tra infiammatorio della parete venosa e del trombo venoso occlusivo. Dalla nostra esperienza, abbiamo scoperto che 10 o più animali per gruppo sono necessari per ottenere una significatività statistica tra i gruppi per i pesi trombo, vena c...

<table border="1"><tbody><tr><td> Materiali di consumo e attrezzature </td></tr><tr><td> Gas isoflurano </td></tr><tr><td> Prolene 7-0 sutura </td></tr><tr><td> Vicryl 5-0 sutura </td></tr><tr><td> Colla pelle </td></tr><tr><td> Occhio lubrificante </td></tr><tr><td> Nolvasan soluzione o la soluzione batteriostatica </td></tr><tr><td> Stereomicroscopio portata </td></tr><tr><td> MediCapture </td></tr><tr><td> Cotone Applicatori </td></tr><tr><td> Garza 2x2 </td></tr><tr><td> Riscaldamento pad </td></tr><tr><td> Anestetico macchina </td></tr><tr><td> Perle di vetro strumento sterilizzatore </td></tr><tr><td> Micro strumenti chirurgici </td></tr><tr><td> Restrainers addominale </td></tr></tbody></table>

mouse complete stasis model of inferior vena cava thrombosis

Tromboembolismo venoso (TEV) comprende sia la trombosi venosa profonda (TVP) ed embolia polmonare (EP). Negli Stati Uniti (US), l&#39;alta morbilità e mortalità tassi di TEV fare un serio problema di salute 1-2. Dopo le malattie cardiache e ictus, tromboembolismo venoso è il terzo 3 più comuni malattie vascolari. Solo negli Stati Uniti, c&#39;è un circa 900.000 persone colpite ogni anno, con 300.000 decessi che si verificano ogni anno 3. Un affidabile in un modello animale in vivo per studiare i meccanismi di questa malattia è necessario. I vantaggi di utilizzare il modello murino completa stasi di trombosi della vena cava inferiore sono diversi. Il modello del mouse permette la somministrazione di volumi molto piccoli di agenti limitata disponibilità di test, riducendo i costi drasticamente. Più promettente è la possibilità di topi con knockout gene che permettono funzioni specifiche fattore infiammatorio e della coagulazione a delinearsi. Attuali test molecolare consentono la quantificazione di parete venosa, trombi, sangue intero e plasma per saggi. Tuttavia, una delle maggiori preoccupazioni che coinvolgono questo modello è il limitare la dimensione operativa e la friabilità dei vasi. Inoltre, a causa del piccolo peso campione IVC (media 0,005 grammi), è necessario aumentare il numero degli animali per l&#39;accurata analisi statistiche per il tessuto, trombo, e saggi di sangue come real-time polymerase chain reaction (RT-PCR), Western Blot, enzyme-linked immunoenzimatico (ELISA), zimografia, parete venosa e trombosi analisi cellulare e sangue intero e plasma saggi 4-8. Il principale svantaggio con il modello di stasi è che la mancanza di flusso di sangue inibisce l&#39;effetto massimo di somministrare agenti terapeutici sistemici sul trombo e la parete venosa.

Watch this Scientific Journal Video about Mouse Complete Stasis Model of Inferior Vena Cava Thrombosis at JoVE.com

Mouse Complete Stasis Model of Inferior Vena Cava Thrombosis

L&#39;intero modello murino stasi di trombosi della vena cava inferiore rendimenti quantificabile quantità di tessuto della parete venosa e trombosi. Si è dimostrato utile per valutare le interazioni tra la parete venosa e il trombo occlusivo e nel valutare la progressione da acuta a cronica.

Modello del mouse completa stasi della vena cava inferiore Trombosi

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

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Research

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Medicine

18.1K Views.  University of Michigan. L'intero modello murino stasi di trombosi della vena cava inferiore rendimenti quantificabile quantità di tessuto della parete venosa e trombosi. Si è dimostrato utile per valutare le interazioni tra la parete venosa e il trombo occlusivo e nel valutare la progressione da acuta a cronica.

Video: Mouse Complete Stasis Model of Inferior Vena Cava Thrombosis

Electrolytic Inferior Vena Cava Model (EIM) of Venous Thrombosis

Animal models serve a vital role in deep venous thrombosis (DVT) research in order to study thrombus formation, thrombus resolution and to test potential therapeutic compounds (1). New compounds to be utilized in the treatment and prevention of DVT are currently being developed. The delivery of potential therapeutic antagonist compounds to an affected thrombosed vein has been problematic. In the context of therapeutic applications, a model that uses partial stasis and consistently generates thrombi within a major vein has been recently established. The Electrolytic Inferior vena cava Model (EIM) is mouse model of DVT that permits thrombus formation in the presence of continuous blood flow. This model allows therapeutic agents to be in contact with the thrombus in a dynamic fashion, and is more sensitive than other models of DVT (1). In addition, this thrombosis model closely simulates clinical situations of thrombus formation and is ideal to study venous endothelial cell activation, leukocyte migration, venous thrombogenesis, and to test therapeutic applications (1). The EIM model is technically simple, easily reproducible, creates consistent thrombi sizes and allows for a large sample (i.e. thrombus and vein wall) which is required for analytical purposes.

Electrolytic Inferior Vena Cava Model EIM of Venous Thrombosis

The electrolytic induction of endothelial activation to the internal surface of the Inferior Vena Cava results in venous type thrombus formation due to endothelial activation and partial blood stasis, two components of Virchow's triad.

Elettrolitico vena cava inferiore Model (EIM) della Trombosi Venosa

Animal models serve a vital role in deep venous thrombosis (DVT) research in order to study thrombus formation, thrombus resolution and to test potential ...

Ricerca

Medicina

Quantitative Analysis and Characterization of Atherosclerotic Lesions in the Murine Aortic Sinus

Atherosclerosis is a disease of the large arteries and a major underlying cause of myocardial infarction and stroke. Several different mouse models have been developed to facilitate the study of the molecular and cellular pathophysiology of this disease. In this manuscript we describe specific techniques for the quantification and characterization of atherosclerotic lesions in the murine aortic sinus and ascending aorta. The advantage of this procedure is that it provides an accurate measurement of the cross-sectional area and total volume of the lesion, which can be used to compare atherosclerotic progression across different treatment groups. This is possible through the use of the valve leaflets as an anatomical landmark, together with careful adjustment of the sectioning angle. We also describe basic staining methods that can be used to begin to characterize atherosclerotic progression. These can be further modified to investigate antigens of specific interest to the researcher. The described techniques are generally applicable to a wide variety of existing and newly created dietary and genetically-induced models of atherogenesis.

We describe procedures to quantify and characterize atherosclerotic lesions in mouse models using precision sectioning of the aortic sinus and ascending aorta combined with histochemical and immunohistochemical analysis.

Atherosclerosis is a disease of the large arteries and a major underlying cause of myocardial infarction and stroke. Several different mouse models have ...

Implantation of Inferior Vena Cava Interposition Graft in Mouse Model

Biodegradable scaffolds seeded with bone marrow mononuclear cells (BMCs) are often used for reconstructive surgery to treat congenital cardiac anomalies. The long-term clinical results showed excellent patency rates, however, with significant incidence of stenosis. To investigate the cellular and molecular mechanisms of vascular neotissue formation and prevent stenosis development in tissue engineered vascular grafts (TEVGs), we developed a mouse model of the graft with approximately 1 mm internal diameter. First, the TEVGs were assembled from biodegradable tubular scaffolds fabricated from a polyglycolic acid nonwoven felt mesh coated with &#949;-caprolactone and L-lactide copolymer. The scaffolds were then placed in a lyophilizer, vacuumed for 24 hr, and stored in a desiccator until cell seeding. Second, bone marrow was collected from donor mice and mononuclear cells were isolated by density gradient centrifugation. Third, approximately one million cells were seeded on a scaffold and incubated O/N. Finally, the seeded scaffolds were then implanted as infrarenal vena cava interposition grafts in C57BL/6 mice. The implanted grafts demonstrated excellent patency (&#62;90%) without evidence of thromboembolic complications or aneurysmal formation. This murine model will aid us in understanding and quantifying the cellular and molecular mechanisms of neotissue formation in the TEVG.

To improve our knowledge of cellular and molecular neotissue formation, a murine model of the TEVG was recently developed. The grafts were implanted as infrarenal vena cava interposition grafts in C57BL/6 mice. This model achieves similar results to those achieved in our clinical investigation, but over a far shortened time-course.

L&#39;impianto di Vena cava inferiore Interposizione Graft in modello murino

Biodegradable scaffolds seeded with bone marrow mononuclear cells (BMCs) are often used for reconstructive surgery to treat congenital cardiac anomalies. ...

Orthotopic Injection of Breast Cancer Cells into the Mammary Fat Pad of Mice to Study Tumor Growth.

Breast cancer growth can be studied in mice using a plethora of models. Genetic manipulation of breast cancer cells may provide insights into the functions of proteins involved in oncogenic progression or help to discover new tumor suppressors. In addition, injecting cancer cells into mice with different genotypes might provide a better understanding of the importance of the stromal compartment. Many models may be useful to investigate certain aspects of disease progression but do not recapitulate the entire cancerous process. In contrast, breast cancer cells engraftment to the mammary fat pad of mice better recapitulates the location of the disease and presence of the proper stromal compartment and therefore better mimics human cancerous disease. In this article, we describe how to implant breast cancer cells into mice orthotopically and explain how to collect tissues to analyse the tumor milieu and metastasis to distant organs. Using this model, many aspects (growth, angiogenesis, and metastasis) of cancer can be investigated simply by providing a proper environment for tumor cells to grow.

Cancer is a complex disease that is influenced by the tissue surrounding the tumor as well as local pro- and anti-inflammatory mediators. Therefore, orthotropic injection models, rather than subcutaneous models may be useful to study cancer progression in a manner that better mimics human pathology.

Iniezione ortotopico di cellule di carcinoma mammario nel mammaria Fat Pad di Mice per studiare la crescita del tumore.

Breast cancer growth can be studied in mice using a plethora of models. Genetic manipulation of breast cancer cells may provide insights into the ...

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.

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.

Cloruro ferrico-indotta Trombosi modello murino sulla carotide e Mesentere 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 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.

Ferrico I modelli murini Trombosi indotta da cloruri

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

Patch Angioplasty in the Rat Aorta or Inferior Vena Cava

Pericardial patches are commonly used in vascular surgery to close vessels. To facilitate studies of the neointimal hyperplasia that forms on the patch, we developed a rat model of patch angioplasty that can be used in either a vein or an artery, creating a patch venoplasty or a patch arterioplasty, respectively. Technical aspects of this model are discussed. The infra-renal IVC or aorta are dissected and then clamped proximally and distally. A 3 mm venotomy or arteriotomy is performed in the infrarenal inferior vena cava or aorta of 6 to 8 week-old Wistar rats. A bovine pericardial patch (3 mm x 1.5 mm x 0.6 mm) is then used to close the site using a 10-0 nylon suture. Compared to arterial patches, venous patches show increased neointimal thickness on postoperative day 7. This novel model of pericardial patch angioplasty can be used to examine neointimal hyperplasia on vascular biomaterials, as well as to compare the differences between the arterial and venous environments.

We have established a model of pericardial patch angioplasty that can be used in either small-diameter veins or arteries. This model can be used to compare venous and arterial neointimal hyperplasia formation.

Patch Angioplastica nel ratto aorta o vena cava inferiore

Pericardial patches are commonly used in vascular surgery to close vessels. To facilitate studies of the neointimal hyperplasia that forms on the patch, ...

A Rat Model of Orthotopic Liver Transplantation Using a Novel Magnetic Anastomosis Technique for Suprahepatic Vena Cava Reconstruction

The rat model of orthotopic liver transplantation (OLT) is essential for transplant research. It is a very sophisticated animal model and requires a steep learning curve. The introduction of the cuff technique for anastomosis of the portal vein (PV) and infrahepatic vena cava (IHVC) has significantly simplified the transplant procedure in rats. However, due to the short anterior wall of the recipients' suprahepatic vena cava (SHVC), the cuff technique is very difficult to use for the reconstruction of the SHVC. Most researchers in this field still use the hand-suture technique for SHVC reconstruction, which makes it the bottleneck step in rat orthotopic liver transplantation. The magnetic anastomosis technique (i.e., magnamosis) is a method of connecting two vessels using the attractive force between two magnets. Our recent study has shown that the magnetic anastomosis technique is superior to the hand-suture technique for SHVC reconstruction in rats. In this article, we show a step-by-step protocol for SHVC reconstruction in rats using the novel magnetic anastomosis technique. In this model, the reconstruction of the PV and IHVC was performed by the standard cuff technique, while the reconstruction of the bile duct (BD) was performed by a stent technique. The hepatic re-arterialization was not performed. The magnetic anastomosis technique made SHVC reconstruction much easier and significantly shortened the anphepatic phase. After a reasonable learning curve, even researchers without advanced microsurgical skills can produce reliable and reproducible results using this rat model of OLT.

The reconstruction of the suprahepatic vena cava (SHVC) remains a difficult step in rat orthotopic liver transplantation. In this article, we show a step-by-step protocol for SHVC reconstruction in rats using a novel magnetic anastomosis technique.

Un modello del ratto di orthotopic del trapianto del fegato usando una tecnica novella dell'anastomosi magnetico per la ricostruzione della Vena Cava Suprahepatic

The rat model of orthotopic liver transplantation (OLT) is essential for transplant research. It is a very sophisticated animal model and requires a steep ...

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.

Trombosi dell'arteria carotica canini cloruro ferrico-indotta: Un grande modello animale di danno vascolare

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

Isometric Contractility Measurement of the Mouse Mesenteric Artery Using Wire Myography

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and drugs. It is widely used in many studies on the physiological functions of vascular smooth muscle, the pathogenesis of vascular diseases such as hypertension, and the development of smooth muscle relaxant drugs. The mouse is a widely used model animal with a large pool of disease models and genetically modified strains. We introduced this method to measure the isometric contraction of mouse mesenteric artery in detail. A 1.4-mm segment of mouse mesenteric resistance artery was isolated and mounted on a myograph chamber by passing two steel wires through its lumen. After equilibration and normalization steps, the vessel segment was potentiated by a high-K+ solution twice prior to the contraction assay. As an example of the application of this method in drug development, we measured the relaxant effect of a novel natural substance, neoliensinine, isolated from a Chinese herb, embryos of the lotus seed (Nelumbo nucifera Gaertn.) on mouse mesenteric arteries. The vessel segments mounted on the myograph chamber were stimulated with a high-K+ solution. When the force tension reached a stable sustained phase, cumulative doses of neoliensinine were added to the chamber. We found that neoliensinine had a dose-dependent relaxant effect on smooth muscle contraction, thus suggesting that it bears potential activity against hypertension. In addition, as the vessel segment can survive at least 4 hours after mounting and maintain contractility induced by the high-K+ solution for many times, we suggest that the wire myograph system may be used for the time-consuming process of drug screening.

The wire myograph technique is used to investigate vascular smooth muscle functions and screen new drugs. We report a detailed protocol for measuring the isometric contractility of the mouse mesenteric artery and for screening new relaxants of vascular smooth muscle.

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

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and ...