Questo lavoro è stato supportato in parte dal National Institutes of Health sovvenzioni R56-HL095498, R01-HL-095.498 (AD), R01-HL128406, il Merit Review Award I01-BX002336 dal Department of Veterans Affairs Biomedical Research Laboratory negli Stati Uniti e Programma di sviluppo (aD), un reparto di Yale di Chirurgia premio Ohse (aD), un Sarnoff Cardiovascular Foundation Fellowship (a JMS), e attraverso le risorse e l&#39;utilizzo delle strutture presso il Connecticut Healthcare System Veterans Affairs (West Haven, CT ). patch di pericardio bovino sono stati generosamente forniti da Lemaitre Vascular Inc. (Burlington, MA).

Approvazione da parte del Comitato di cura e l&#39;uso di animali istituzionale adeguato deve essere ottenuto, che indica che il protocollo segue le linee guida del comitato etico di ricerca animale del proprio istituto. 1. Anestesia e procedure pre-operatorie <ol><li> Anestetizzare ratti maschi Wistar, del peso di circa 200 g, con vaporizzato 3% isoflurano e 1 L / min di ossigeno somministrato in una camera di induzione acrilico. </li><li> Confermare un&#39;anestesia adeguata per mancanza di reazione ad un dito del piede e la coda pizzico. Posizionare pomata sugli occhi per prevenire la secchezza mentre gli animali sono sotto anestesia. Posizionare la supina ogni ratto sulla piastra di funzionamento di plastica e posizionare una maschera di silicone per fornire vaporizzato 2 - 3% isoflurano per inalazione continua. Non lasciare un animale incustodito fino a quando non ha ripreso conoscenza sufficiente a mantenere decubito sternale. </li><li> Indossando guanti sterili, rimuovere i capelli ventrale utilizzando un dispositivo di rimozione dei capelli. Buprenorphine, sottocutanea o intraperitoneale, è spesso data a questo punto. </li></ol> 2. Procedure Operative <ol><li> Patch venoplasty <ol><li> Prima della chirurgia, tagliare la patch di pericardio bovino utilizzando un microscissor a una dimensione tre millimetri x 1,5 mm x 0,6 mm. Immergere il cerotto in 10 mL di soluzione salina sterile per 5 minuti in un piatto di coltura cellulare. </li><li> Preparare il sito di incisione con un antisettico topico secondo il protocollo istituzionale e applicare un telo chirurgico. Utilizzare guanti e strumenti sterili per mantenere la tecnica asettica in tutta la chirurgia. L&#39;anestesia locale possono essere iniettati nel sito della incisione cutanea previsto. </li><li> Usando un bisturi, fare una incisione addominale mediana estende dal livello del xiphoid appena sopra il pube. </li><li> Inserire un divaricatore e sviscerare tutte le viscere dalla cavità addominale verso il lato destro. Avvolgere le viscere in una garza imbevuta con soluzione fisiologica. </li><li> Esporre l&#39;ivena cava nfrarenal (IVC). Il sito del cerotto dell&#39;impianto è di circa 2 mm sotto il livello dell&#39;origine delle vene renali; sezionare il IVC gratuitamente in questo sito. Legare e dividere tutte le vene lombari e tributari a questo livello con 6-0 punti di sutura in nylon. </li><li> Mettere due clip di microchirurgia attraverso sia prossimale e distale infarenal IVC. Eseguire un venotomia con un ago 25 G perforando la parete anteriore della IVC, e poi estenderla a circa 3 mm con forbici (Figura 1A). </li><li> Chiudere il venotomia con la patch utilizzando in esecuzione o interrotto 10-0 suture in nylon (Figura 1B). </li><li> Completare la chiusura venoplasty. Rimuovere il morsetto distale prima di sfogare la patch e quindi rimuovere il morsetto prossimale per ripristinare il flusso di sangue al IVC. </li><li> Confermare emostasi osservando per 30 s senza compressione. Riportare le viscere alla loro posizione naturale. Chiudere l&#39;addome con una sutura in esecuzione secondo protocolli approvati animali. </li></ol></li><li> Patch Angioplastica <ol><li> Prima della chirurgia, tagliare la patch di pericardio bovino utilizzando un microscissor a una dimensione tre millimetri x 1,5 mm x 0,6 mm. Immergerlo in soluzione fisiologica sterile. </li><li> Preparare il sito di incisione con un antisettico topico e applicare un telo chirurgico. Utilizzare guanti e strumenti sterili per mantenere la tecnica asettica in tutta la chirurgia. </li><li> Usando un bisturi, fare una incisione addominale mediana estende dal livello del xiphoid appena sopra il pube. </li><li> Inserire un divaricatore e sviscerare tutte le viscere dalla cavità addominale verso il lato destro. Avvolgere le viscere in una garza imbevuta con soluzione fisiologica. </li><li> Esporre l&#39;aorta. Il sito del cerotto dell&#39;impianto è di circa 2 mm sotto il livello dell&#39;origine delle arterie renali; sezionare l&#39;aorta gratuitamente in questo sito. Legare e dividere tutte le arterie lombari a questo livello con 6-0 punti di sutura in nylon. </li><li> Bloccare l&#39;aorta sottorenale. Utilizzare un ago G 25 per eseguire un arteriotomia da puncturing parete anteriore dell&#39;aorta, e poi estenderla a circa 3 mm forbici usando (Figura 1C). </li><li> Chiudere il arteriotomia con la patch utilizzando in esecuzione 10-0 suture in nylon (Figura 1D). Completare la chiusura Angioplastica. Rimuovere il morsetto distale prima di sfogare la patch e quindi rimuovere il morsetto prossimale per ripristinare il flusso di sangue all&#39;aorta. </li><li> Confermare emostasi osservando per 30 s senza compressione. Riportare le viscere alla loro posizione naturale. Chiudere l&#39;addome con una sutura in esecuzione secondo protocolli approvati animali. </li></ol></li></ol> Procedure 3. post-operatorie <ol><li> Dopo la chiusura del ventre, interrompere l&#39;anestesia. Applicare cure post-operatorie, tra cui l&#39;analgesia e la cura delle ferite, secondo le istruzioni raccomandate dal Comitato Cura e uso istituzionale degli animali. </li><li> Per l&#39;analgesia, utilizzare buprenorfina a 0,1 mg / kg per via intramuscolare non meno di ogni 12 ore per 24 ore seguonoing le procedure chirurgiche. Controllare lo stato dell&#39;animale ogni giorno nella sala animale, garantendo il corretto recupero dal periodo perioperatorio nonché un&#39;adeguata trattamento del dolore post-operatorio. Non restituire un animale che ha subito un intervento chirurgico per la compagnia di altri animali fino a quando non ha pienamente recuperato. </li></ol><style type="text/css">p.p1 {margin: 0.0px 0.0px 0.0px 0.0px; font: 12.0px Helvetica}
</style><style type="text/css">p.p1 {margin: 0.0px 0.0px 0.0px 0.0px; font: 12.0px Helvetic
</style>

<ol>
	<li>Muto, A., Nishibe, T., Dardik, H., Dardik, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Patches+for+carotid+artery+endarterectomy:+current+materials+and+prospects.">Patches for carotid artery endarterectomy: current materials and prospects.</a> J Vasc Surg. 50 (1), 206-213 (2009).</li><li>Li, X., 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=Current+usage+and+future+directions+for+the+bovine+pericardial+patch.">Current usage and future directions for the bovine pericardial patch.</a> Ann Vasc Surg. 25 (4), 561-568 (2011).</li><li>Biasi, G. M., Sternjakob, S., Mingazzini, P. M., Ferrari, S. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Nine-year+experience+of+bovine+pericardium+patch+angioplasty+during+carotid+endarterectomy.">Nine-year experience of bovine pericardium patch angioplasty during carotid endarterectomy.</a> J Vasc Surg. 36 (2), 271-277 (2002).</li><li>Papakostas, 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=Use+of+the+vascu-guard+bovine+pericardium+patch+for+arteriotomy+closure+in+carotid+endarterectomy.+Early+and+long-term+results.">Use of the vascu-guard bovine pericardium patch for arteriotomy closure in carotid endarterectomy. Early and long-term results.</a> Ann Vasc Surg. 28 (5), 1213-1218 (2014).</li><li>Ohwada, 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=Glutaraldehyde-fixed+heterologous+pericardium+for+vena+cava+grafting+following+hepatectomy.">Glutaraldehyde-fixed heterologous pericardium for vena cava grafting following hepatectomy.</a> Hepatogastroenterology. 46 (26), 855-858 (1999).</li><li>Mori, 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=Standard+hepatic+vein+reconstruction+with+patch+plasty+using+the+native+portal+vein+in+adult+living+donor+liver+transplantation.">Standard hepatic vein reconstruction with patch plasty using the native portal vein in adult living donor liver transplantation.</a> Liver Transpl. 18 (5), 602-607 (2012).</li><li>Bai, 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=Pericardial+patch+venoplasty+heals+via+attraction+of+venous+progenitor+cells.">Pericardial patch venoplasty heals via attraction of venous progenitor cells.</a> Physiological Reports. 4 (12), (2016).</li><li>Li, X., 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=Pericardial+patch+angioplasty+heals+via+an+Ephrin-B2+and+CD34+positive+cell+mediated+mechanism.">Pericardial patch angioplasty heals via an Ephrin-B2 and CD34 positive cell mediated mechanism.</a> PLoS One. 7 (6), 38844 (2012).</li><li>Bai, 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=Pretreatment+of+pericardial+patches+with+antibiotics+does+not+alter+patch+healing+in+vivo.">Pretreatment of pericardial patches with antibiotics does not alter patch healing in vivo.</a> J Vasc Surg. 63 (4), 1063-1073 (2016).</li><li>Muto, 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=Eph-B4+prevents+venous+adaptive+remodeling+in+the+adult+arterial+environment.">Eph-B4 prevents venous adaptive remodeling in the adult arterial environment.</a> J Exp Med. 208 (3), 561-575 (2011).</li></ol>

The authors have nothing to disclose.

Descriviamo un modello angioplastica patch nel topo che può essere utilizzato sia in vene o arterie, creando un venoplasty patch o patch di Angioplastica rispettivamente. Utilizzando questo modello di patch pericardico, abbiamo mostrato aggressivo e stabile iperplasia neointimale nel ratto IVC venoplasty rispetto al Angioplastica aortico (Figura 2) 7, 8. Diversi aspetti tecnici sono notevoli. Nel Angioplastica aorti...

Chirurghi vascolari comunemente stretti vasi sanguigni con patch pericardico, sia bovina o suina, per ridurre il rischio di restenosi 1, 2. Nella chirurgia arteriosa, pericardico cerotto angioplastica riduce il tasso di restenosi rispetto ad una chiusura primaria 3, 4. Patch venoplasty è anche simile eseguita per chiudere le vene, anche se meno frequentemente rispetto alle procedure arteriosa 5, 6. Le patch di solito si sviluppano neointimale sulla loro superficie luminale; Tuttavia, il meccanismo di formazione della neointima è chiaro, soprattutto nel confronto delle differenze in formazione neointimale tra arterioso e venoso ambienti. Abbiamo precedentemente dimostrato che le patch pericardico attraggono le cellule progenitrici sia arterioso e venoso degli ambienti e quella patch venoso angioplastica mostra più di unaformazione neointimale ggressive rispetto ai cerotti arteriosi 7, 8. In questo studio, descriviamo il modello di ratto di angioplastica patch di pericardio che è comune a chiudere entrambe le vene e le arterie e che possono essere utilizzati per studiare la formazione neointimale; Questo metodo è un piccolo modello animale di una tecnica operatoria comunemente utilizzato in pazienti umani. Il modello di ratto ha un diametro maggiore del modello di topo, facilitando aspetti tecnici della chirurgia e migliorando la curva di apprendimento per il modello.

<table border="0" cellpadding="0" cellspacing="0" width="523">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Bovine pericardial patch</td>
			<td colspan="2" style="width:307px;">LeMaitre Vascular</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Vascular clamp</td>
			<td>Roboz Surgical Instrument Co.</td>
			<td>RS-5424</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Clamp applying forceps</td>
			<td>Roboz Surgical Instrument Co</td>
			<td>RS-5410</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">10-0 Prolene suture</td>
			<td>Arosurgical Co.</td>
			<td>T05A10N10-13</td>
		</tr>
	</tbody>
</table>

patch angioplasty in the rat aorta or inferior vena cava

patch pericardiche sono comunemente utilizzati in chirurgia vascolare per chiudere i vasi. Per facilitare gli studi di iperplasia neointimale che si forma sulla patch, abbiamo sviluppato un modello di ratto di angioplastica cerotto che può essere utilizzato sia in una vena o un&#39;arteria, creando un venoplasty patch o un Angioplastica patch, rispettivamente. Gli aspetti tecnici di questo modello sono discussi. L&#39;IVC o dell&#39;aorta infra-renali sono sezionati e poi serrati prossimale e distale. Un venotomia 3 mm o arteriotomy viene eseguita in infrarenale vena cava inferiore o aorta da 6 a 8 settimane di età ratti Wistar. Una patch di pericardio bovino (3 mm x 1,5 mm x 0,6 millimetri) viene quindi utilizzato per chiudere il sito utilizzando una sutura 10-0 nylon. Rispetto alle patch arteriose, venose patch mostrano un aumento dello spessore neointimale su postoperatoria giorno 7. Questo nuovo modello di angioplastica cerotto pericardio può essere utilizzato per esaminare neointimale sui biomateriali vascolari, nonché di confrontare le differenze tra arterioso e venosoambienti.

Dopo l&#39;angioplastica patch, non c&#39;era trombosi sia venoso (n = 42) o arteriosa gruppi di patch (n = 24). La sopravvivenza al giorno 7 è stata del 100% in entrambi i gruppi sia con un venosa o arteriosa cerotto. I cerotti sono stati raccolti il giorno 7 e colorate con ematossilina ed eosina, e lo spessore neointimale stato confrontato come precedentemente descritto 7. A causa della differenza morfologica net...

Watch this Scientific Journal Video about Patch Angioplasty in the Rat Aorta or Inferior Vena Cava at JoVE.com

Patch Angioplasty in the Rat Aorta or Inferior Vena Cava

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, ...

patch-angioplasty-in-the-rat-aorta-or-inferior-vena-cava

Research

JoVE Journal

Medicine

10.5K Views.  Yale University. 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.

Video: Patch Angioplasty in the Rat Aorta or Inferior Vena Cava

Hyperinsulinemic-Euglycemic Clamp in the Conscious Rat

Type 2 diabetes (T2D) is rapidly rising in prevalence. Characterized by either inadequate insulin production or the inability to utilize insulin produced, T2D results in elevated blood glucose levels. The "gold-standard" in assessing insulin sensitivity is a hyperinsulinemic-euglycemic clamp or insulin clamp. In this procedure, insulin is infused at a constant rate resulting in a drop in blood glucose. To maintain blood glucose at a constant level, exogenous glucose (D50) is infused into the venous circulation. The amount of glucose infused to maintain homeostasis is indicative of insulin sensitivity. Here, we show the basic clamp procedure in the chronically catheterized, unrestrained, conscious rat. This model allows blood to be collected with minimal stress to the animal. Following the induction of anesthesia, a midline incision is made and the left common carotid artery and right jugular vein are catheterized. Inserted catheters are flushed with heparinized saline, then exteriorized and secured. Animals are allowed to recover for 4-5 days prior to experiments, with weight gain monitored daily. Only those animals who regain weight to pre-surgery levels are used for experiments. On the day of the experiment, rats are fasted and connected to pumps containing insulin and D50. Baseline glucose is assessed from the arterial line and used a benchmark throughout the experiment (euglycemia). Following this, insulin is infused at a constant rate into the venous circulation. To match the drop in blood glucose, D50 is infused. If the rate of D50 infusion is greater than the rate of uptake, a rise in glucose will occur. Similarly, if the rate is insufficient to match whole body glucose uptake, a drop will occur. Titration of glucose continues until stable glucose readings are achieved. Glucose levels and glucose infusion rates during this stable period are recorded and reported. Results provide an index of whole body insulin sensitivity. The technique can be refined to meet specific experimental requirements. It is further enhanced by the use of radioactive tracers that can determine tissue specific insulin-stimulated glucose uptake as well as whole body glucose turnover.

The hyperinsulinemic-euglycemic clamp is the "gold standard" for the assessment of insulin action. Insulin is infused at a constant rate stimulating glucose uptake. The amount of exogenous glucose infused to counter this drop is indicative of insulin sensitivity. Here the procedure is performed on a conscious, unrestrained rat.

Iperinsulinemico-euglicemico pinza nei topi

Type 2 diabetes (T2D) is rapidly rising in prevalence. Characterized by either inadequate insulin production or the inability to utilize insulin produced, ...

Ricerca

Medicina

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 ...

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.

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 ...

Angiogenesis in the Ischemic Rat Lung

The adult lung is perfused by both the systemic bronchial artery and the entire venous return flowing through the pulmonary arteries. In most lung pathologies, it is the smaller systemic vasculature that responds to a need for enhanced lung perfusion and shows robust neovascularization. Pulmonary vascular ischemia induced by pulmonary artery obstruction has been shown to result in rapid systemic arterial angiogenesis in man as well as in several animal models. Although the histologic assessment of the time course of bronchial artery proliferation in rats was carefully described by Weibel 1, mechanisms responsible for this organized growth of new vessels are not clear. We provide surgical details of inducing left pulmonary artery ischemia in the rat that leads to bronchial neovascularization. Quantification of the extent of angiogenesis presents an additional challenge due to the presence of the two vascular beds within the lung. Methods to determine functional angiogenesis based on labeled microsphere injections are provided.

The lung is perfused by both the systemic bronchial artery and pulmonary arteries. In most lung pathologies, it is the smaller systemic vasculature that shows robust neovascularization. Cessation of pulmonary blood flow promotes brisk bronchial angiogenesis. We provide surgical details of inducing left pulmonary artery ischemia that promotes bronchial neovascularization.

L&#39;angiogenesi nel polmone del ratto ischemica

The  adult lung is perfused by both the systemic bronchial artery and the entire  venous return flowing through the pulmonary arteries. In most lung ...

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. ...

Scleral Cross-linking Using Riboflavin and Ultraviolet-A Radiation for Prevention of Axial Myopia in a Rabbit Model

Myopic individuals, especially those with severe myopia, are at higher-than-normal risk of cataract, glaucoma, retinal detachment and chorioretinal abnormalities. In addition, pathological myopia is a common irreversible cause of visual impairment and blindness1-3. Our study demonstrates the effect of scleral crosslinking using riboflavin and ultraviolet-A radiation on the development of axial myopia in a rabbit model. The axial length of the eyeball was measured by A-scan ultrasound in New Zealand white rabbits aged 13 days (male and female). The eye then underwent 360&#176; conjunctival peritomy with scleral crosslinking, followed by tarsorrhaphy. Axial elongation was induced in 13 day-old New Zealand rabbits by suturing their right eye eyelids (tarsorrhaphy). The eyes were divided into quadrants, and every quadrant had two scleral irradiation zones, each with an area of 0.2 cm&#178; and a radius of 4 mm. Crosslinking was performed by dropping 0.1% dextran-free riboflavin-5-phosphate onto the irradiation zones 20 sec before ultraviolet-A irradiation and every 20 sec during the 200 sec irradiation time. UVA radiation (370 nm) was applied perpendicular to the sclera at 57 mW/cm&#178; (total UVA light dose, 57 J/cm&#178;). Tarsorrhaphies were removed on day 55, followed by repeated axial length measurements. This study demonstrates that scleral crosslinking with riboflavin and ultraviolet-A radiation effectively prevents occlusion-induced axial elongation in a rabbit model.

We demonstrate the effect of scleral crosslinking with riboflavin and UVA on an axial elongation rabbit eye. Axial elongation was induced in 13 day-old New Zealand rabbits (male and female) by suturing their right eye eyelids (tarsorrhaphy).

Sclerale reticolazione Utilizzando riboflavina e raggi ultravioletti-A di radiazione per la prevenzione della miopia assiale in un modello di coniglio

Myopic individuals, especially those with severe myopia, are at higher-than-normal risk of cataract, glaucoma, retinal detachment and chorioretinal ...

A Model of Cardiac Remodeling Through Constriction of the Abdominal Aorta in Rats

Heart failure is one of the leading causes of death worldwide. It is a complex clinical syndromethat includes fatigue, dyspnea, exercise intolerance, and fluid retention. Changes in myocardial structure, electrical conduction, and energy metabolism develop with heart failure, leading to contractile dysfunction, increased risk of arrhythmias, and sudden death. Hypertensive heart disease is one of the key contributing factors of cardiac remodeling associated with heart failure. The most commonly-used animal model mimicking hypertensive heart disease is created via surgical interventions, such as by narrowing the aorta. Abdominal aortic constriction is a useful experimental technique to induce a pressure overload, which leads to heart failure. The surgery can be easily performed, without the need for chest opening or mechanical ventilation. Abdominal aortic constriction-induced cardiac pathology progresses gradually, making this model relevant to clinical hypertensive heart failure. Cardiac injury and remodeling can be observed 10 weeks after the surgery. The method described here provides a simple and effective approach to produce a hypertensive heart disease animal model that is suitable for studying disease mechanisms and for testing novel therapeutics.

A rat model of abdominal aortic constriction that induces cardiac hypertrophy and remodeling is described. An efficient, highly-reproducible, and minimally-invasive method is used to provide a simple yet useful platform for research in myocardial hypertrophy and dysfunction.

Un modello di rimodellamento cardiaco Attraverso Costrizione del Aorta addominale in Rats

Heart failure is one of the leading causes of death worldwide. It is a complex clinical syndromethat includes fatigue, dyspnea, exercise intolerance, and ...

Balloon-based Injury to Induce Myointimal Hyperplasia in the Mouse 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 balloon denudation model, which is comparable with established vessel injury models in large animals. The aorta denudation model with balloon catheters mimics the clinical setting and leads to comparable pathobiological and physiological changes. Briefly, after performing a horizontal incision in the aorta abdominalis, a balloon catheter will be inserted into the vessel, inflated, and introduced retrogradely. Inflation of the balloon will lead to intima injury and overdistension of the vessel. After removing the catheter, the aortic incision will be closed with single stiches. The model shown in this article is reproducible, easy to perform, and can be established quickly and reliably. It is especially suitable for evaluating expensive experimental therapeutic agents, which can be applied in an economical fashion. By using different knockout-mouse strains, the impact of different genes on MH development can be assessed.

This article demonstrates a murine model to study the development of myointimal hyperplasia (MH) after aortic balloon injury.

Basato su palloncino ferita per indurre iperplasia Myointimal nell'Aorta addominale di Mouse

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 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 ...

Ultrasound Imaging of the Thoracic and Abdominal Aorta in Mice to Determine Aneurysm Dimensions

Contemporary high-resolution ultrasound instruments have sufficient resolution to facilitate the measurement of mouse aortas. These instruments have been widely used to measure aortic dimensions in mouse models of aortic aneurysms. Aortic aneurysms are defined as permanent dilations of the aorta, which occur most frequently in the ascending and abdominal regions. Sequential measurements of aortic dimensions by ultrasound are the principal approach for assessing the development and progression of aortic aneurysms in vivo. Although many reported studies used ultrasound imaging to measure aortic diameters as a primary endpoint, there are confounding factors, such as probe position and cardiac cycle, that may impact the accuracy of data acquisition, analysis, and interpretation. The purpose of this protocol is to provide a practical guide on the use of ultrasound to measure the aortic diameter in a reliable and reproducible manner. This protocol introduces the preparation of mice and instruments, the acquisition of appropriate ultrasound images, and data analysis.

Ultrasound imaging has become a common modality to determine the luminal dimensions of thoracic and abdominal aortic aneurysms in mice. This protocol describes the procedure to acquire reliable and reproducible two-dimensional ultrasound images of the ascending and abdominal aorta in mice.

Formazione immagine di ultrasuono dell'Aorta toracica ed addominale in topi per determinare le dimensioni di Aneurysm

Contemporary high-resolution ultrasound instruments have sufficient resolution to facilitate the measurement of mouse aortas. These instruments have been ...

Patch Angioplastica nel ratto aorta o vena cava inferiore

Riepilogo

Esplora Altri Video

Capitoli in questo video