Gli autori ringraziano Christiane Pahrmann per l&#39;assistenza tecnica. SS ha ricevuto finanziamenti dalla Deutsche Forschungsgemeinschaft (DFG) (SCHR992/3-2 e SCHR992/4-2).

Tutti i lavori degli animali deve essere effettuata secondo le pertinenti linee guida per la cura degli animali. Ottenere l&#39;approvazione istituzionale per il lavoro animale prima dell&#39;inizio del protocollo. 1. Miointimale Iperplasia Model (A) Per il modello iperplasia intimale, acquistare C57BL/6J (numero Archivio 000.664, C57BL/6J) topi all&#39;età di otto settimane del peso di circa 25 g. Topi Casa in questi esperimenti in condizioni convenzionali; nutrire topi chow mouse standard e fornire acqua ad libitum. <ol><li> Preparazione Mouse: Introdurre il mouse per uno stato anestetico con isoflurano (2%) utilizzando una camera di induzione. <ol><li> Rimuovere i capelli addominale utilizzando un trimmer capelli, gettare il mouse sulla sua schiena e coprire il naso e la bocca con la maschera facciale per assicurare la anestesia. </li><li> Con Provo-Iodio, disinfettare la zona addominale universalmente, seguito dal 80% di etanolo, in due cicli. </li><li> Garantire l&#39;assenzadei riflessi pizzicando dell&#39;arto posteriore per controllare la profondità sufficiente dell&#39;anestesia. </li><li> Separare la pelle ei muscoli lungo la linea alba in due fasi per esporre gli organi addominali. </li><li> Posare l&#39;intestino in un guanto salina idratata. Avvolgere il guanto intorno l&#39;intestino per mantenerli umidi. </li><li> Attentamente sgombrare il tessuto adiposo pezzo per pezzo l&#39;aorta addominale. Esporre l&#39;aorta sottorenale fino alla sua biforcazione, facendo attenzione a non danneggiare alcuna navi filiali. </li><li> Innanzitutto, collocare un morsetto microchirurgico sul prossimale, sottorenale per arrestare il flusso di sangue. Ora, applicare un secondo morsetto distale vicino alla biforcazione aortica. </li><li> Quando il flusso di sangue viene interrotto, fare una piccola incisione nell&#39;aorta vicino al morsetto prossimale. </li><li> Inserire il catetere a palloncino con punta, che è stata idratata con 0,9% di soluzione salina, e avanzare verso il morsetto distale dell&#39;aorta. </li><li> Per il danno endoteliale, denudare l&#39;aorta gonfiando il Balloon lentamente e tirando il catetere nella direzione del morsetto prossimale. </li><li> Ripetere la manovra denudazione due volte. </li><li> Rimuovere il catetere e chiudere l&#39;incisione aortica utilizzando 10-0 prolene suture in esecuzione. </li><li> Aprire con cautela il morsetto distale. In caso di sanguinamento nel sito aortotomy, chiudere il morsetto di nuovo, individuare l&#39;emorragia e mettere ulteriori punti interrotti, se necessario. </li><li> Se nessun sanguinamento può essere osservato, aprire lentamente il morsetto prossimale. </li><li> Un impulso aortico deve essere visibile distalmente dall&#39;incisione. Ora gli intestini possono essere organizzati indietro in situ. </li><li> Sciacquare la cavità addominale utilizzando soluzione fisiologica sterile pre-riscaldato. </li><li> Chiudere la parete addominale che inizia con lo strato muscolare con 6-0 punti di sutura prolene esecuzione. </li><li> Accanto adeguare la pelle sotto l&#39;uso di 5-0 prolene eseguire suture in esecuzione. </li><li> Durante il tempo il mouse è ancora anestetizzato, applicare 4-5 mg / kg Carprofene tramite un inje sottocutaneaction. </li><li> Aggiungi Metamizol l&#39;acqua potabile (50 mg Metamizol per 100 ml) per il controllo del dolore e proseguire per tre giorni dopo l&#39;intervento chirurgico. Controllo degli animali durante il ricovero. Nota: Il periodo di osservazione per questo modello è di 28 giorni. </li></ol></li></ol> 2. Aterosclerosi Model (B) Per il modello aterosclerosi, l&#39;acquisto di apolipoproteina E-deficienti (ApoE - / -) topo (numero Archivio 002.052, B6.129P2-Apoetm1Unc / J) ad un&#39;età di 4 settimane, e poi alimentato per 4-6 mesi. Topi Casa in questi esperimenti in condizioni convenzionali; nutrire topi dieta ad alto contenuto di lipidi occidentale all&#39;arrivo (Harlan Laboratories TD.88137) e fornire acqua ad libitum per una durata di 4-6 mesi. <ol><li> Preparazione Mouse: <ol><li> Feed the ApoE - / - topi con dieta occidentale con inizio alle 4 settimane di età. Manipolazione alimentare deve iniziare direttamente all&#39;arrivo. </li><li> Mantenere la dieta occidentale per tutta la durata dell&#39;esperimento. &lt;/ Li&gt; </ol></li></ol> 3. Analisi <ol><li> Modello iperplasia miointimale (A) La regione di interesse in questo modello è la &quot;zona lesione&quot;. Opzioni per analisi comprendono obliterazione del lume, rapporto I / M (rapporto intima / media), spessore myointima massimo, zona miointimale, Colorazione tricromica di fibrosi e colorazione H &amp; E per cellule mononucleari (come illustrato nelle Figure 1A-1C). Secondo la domanda scientifica numerose altre colorazioni come Picrosirius rosso per il collagene, immunoistochimica e metodi confocale possono essere applicati. </li><li> Modello di aterosclerosi (B) Le regioni di interesse in questo modello sono la valvola aortica, arco aortico e l&#39;aorta discendente. Opzioni per l&#39;analisi: il Sudan colorazione rosso per i lipidi e la colorazione tricromica di valvola aortica, arco aortico e aorta discendente (come illustrato in Fig. 1D). Questo modello produce anche una varietà di altre opzioni come colorazioneper esempio Oil Red O di lipidi neutri, Picrosirius rosso, immunoistochimica e metodi confocale. <ol><li> Sudan colorazione rosso: Eseguire la colorazione rossa Sudan sul tessuto aortico appena raccolte. <ol><li> Rimuovere la 6h dieta prima della raccolta. </li><li> Introdurre il mouse per uno stato anestetico con isoflurano (2%) utilizzando una camera di induzione. Garantire l&#39;assenza di riflessi pizzicando dell&#39;arto posteriore per controllare la profondità sufficiente dell&#39;anestesia. </li><li> Aprite la cassa; tagliare la punta del cuore e perfusione attraverso il ventricolo sinistro, prima con soluzione salina (3 ml), seguito da 4% PFA (3 ml) attraverso la radice aortica per fissare l&#39;aorta. </li><li> Liberare l&#39;aorta dalla radice alla sua biforcazione. Prova per rimuovere il tessuto adiposo, per quanto possibile. </li><li> Tagliare il cuore e la radice aortica, e memorizzarli nel 4% PFA per istologia. </li><li> Per l&#39;istologia della radice aortica deve essere attentamente collocato nel blocco di paraffina in posizione verticale per permettere il taglio della aortic diapositive di generazione radice che mostrano tutte e 3 lembi valvolari. </li><li> Tagliare un&#39;estremità dell&#39;albero aortica e fissarlo sullo strato di silicone con perni sottili. Aprire l&#39;aorta longitudinalmente e fissare il tessuto aortico. </li><li> Lavare il tessuto con il 50% di etanolo. Immergere il tessuto con la soluzione colorante Sudan III per circa 15 min. </li><li> Risciacquare leggermente con il 50% di etanolo per rimuovere l&#39;eccesso Sudan III. (Nota: il 50% di etanolo può lavare via il colorante.) Sciacquare con dH 2 O, e scattare foto per ulteriori analisi. </li></ol></li></ol></li></ol>

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M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Nitro-fatty+acids+reduce+atherosclerosis+in+apolipoprotein+E-deficient+mice.">Nitro-fatty acids reduce atherosclerosis in apolipoprotein E-deficient mice.</a> Arterioscler Thromb Vasc Biol. 30, 938-9345 (2010).</li><li>Rondelet, B., Kerbaul, F., Motte, 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=Bosentan+for+the+prevention+of+overcirculation-induced+experimental+pulmonary+arterial+hypertension.">Bosentan for the prevention of overcirculation-induced experimental pulmonary arterial hypertension.</a> Circulation. 107, 1329-1335 (2003).</li><li>Zeibig, S., Li, Z., Wagner, 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=Effect+of+the+oxLDL+binding+protein+Fc-CD68+on+plaque+extension+and+vulnerability+in+atherosclerosis.">Effect of the oxLDL binding protein Fc-CD68 on plaque extension and vulnerability in atherosclerosis.</a> Circ Res. 108, 695-703 (2011).</li><li>Painter, T. 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Sebbene entrambe le malattie provocano sintomi simili nel paziente, i meccanismi alla base dello sviluppo della placca e quindi i metodi di trattamento sono molto diversi 2. In diverse forme di stenosi arteriosa risultati clinici nei pazienti nonché i tempi di sviluppo placca dipendono dal meccanismo sottostante. A seconda dei risultati clinici nel paziente, diversi metodi di analisi dei campioni devono essere eseguite 5,15. È necessario adattare l&#39;analisi ai rili...

Nei paesi industrializzati la cardiopatia ischemica rimane una delle principali cause di mortalità ad 1. Le cause di stenosi dell&#39;arteria coronaria sono molteplici e includono iperplasia miointimale così come aterosclerosi, con rivascolarizzazione rimanendo la strategia di trattamento più comune 2. Modelli di ricerca che distinguono chiaramente tra le vie meccanicistiche di iperplasia intimale e aterosclerosi sono essenziali per indagare il patobiologico e processi fisiopatologici in arterioso sviluppo della placca. In questo video vengono introdotti due modelli murini diversi utilizzati per studiare sia lo sviluppo di iperplasia miointimale o aterosclerosi. Iperplasia miointimale è postulata per formare tramite il paradigma &quot;risposta a lesioni&quot;, originariamente descritta per l&#39;aterosclerosi 3. La distruzione meccanica dello strato endoteliale porta ad un rimodellamento intenso arricchito da effetti paracrini. Ci sono diversi dimodelli animali fferent comunemente utilizzati per studiare iperplasia miointimale. Alcuni gruppi usano dispositivi metallici in aorta ascendente di ratti 4. Il modello denudazione aortica eseguita con un catetere a palloncino viene anche comunemente usato nei ratti di laboratorio 5,6. Il modello di iperplasia intimale eseguita nei topi di laboratorio 7 implementa legatura della carotide e induce lesioni miointimale 8. Nel nostro laboratorio, il modello denudamento aortico addominale - per studiare lo sviluppo di iperplasia miointimale - così come un modello umanizzato stent restenosi 9 sono comunemente usati. Questo video sottolinea che la scelta del corretto modello sperimentale animale è di fondamentale importanza per gli studi meccanicistici o fisiopatologiche di stenosi arteriosa. Modelli iperplasia miointimale devono essere distinti dai modelli aterosclerosi. Per questi ultimi, apolipoproteina E-deficienti (ApoE - / -) topi in combinazione con dieta occidentale sono comunemente usati per indurre atheroscllesioni erotici 10,11,12,13. Esempi per l&#39;induzione di entrambi questi tipi di malattie miointimale in topi sono mostrati qui, insieme con differenti opzioni del modello adattato per analizzare stenosi 14.

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			<td height="17" style="height:17px;width:156px;">Name</td>
			<td style="width:183px;">Company</td>
			<td style="width:139px;">Catalog number</td>
			<td style="width:140px;">Comments</td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Thyoglycollate Broth 3%</td>
			<td>Fluka</td>
			<td align="right">70157</td>
			<td>powder</td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">PFA 4%</td>
			<td>Electron Microscopy Sciences</td>
			<td>#157135S</td>
			<td align="right">20%</td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Sudan III staining solution</td>
			<td>Sigma Aldrich</td>
			<td>S4131</td>
			<td>powder</td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">mouse C57BL/6J</td>
			<td>Jackson Laboratories</td>
			<td>&#160;Stock # 0006664</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">mouse ApoE-/-</td>
			<td>Jackson Laboratories</td>
			<td>&#160;Stock #002052</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Western Diet</td>
			<td>Harlan Laboratories</td>
			<td>TD.88137</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">hair clipper</td>
			<td>WAHL</td>
			<td>8786-451A ARCO SE</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Forene</td>
			<td>Abbott</td>
			<td></td>
			<td>Isoflurane</td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">microsurgical clamp</td>
			<td>Fine Science Tools</td>
			<td>18055-04</td>
			<td>Micro-Serrefine - 4mm</td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">clamp applicator</td>
			<td>Fine Science Tools</td>
			<td>18056-14</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">catheter</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">10-0 prolene suture</td>
			<td>Ethicon</td>
			<td>788G</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">6-0 prolene suture</td>
			<td>Ethicon</td>
			<td>8709H</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">5-0 prolene suture</td>
			<td>Ethicon</td>
			<td>EH7229H</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Rimadyl</td>
			<td>Pfizer&#160;</td>
			<td></td>
			<td>Carprofen</td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Metacam 1.5mg/ml</td>
			<td>Boehringer Ingelheim</td>
			<td></td>
			<td>Metamizol</td>
		</tr>
	</tbody>
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inducing myointimal hyperplasia versus atherosclerosis in mice: an introduction of two valid models

Vari vivo modelli di roditori di laboratorio in per l&#39;induzione di stenosi sono state stabilite per simulare malattie che comprendono la formazione di placca arteriosa e stenosi, come osservato ad esempio cardiopatia ischemica. Due modelli del mouse altamente riproducibili - sia conseguente stenosi ma ognuno alla base di un diverso percorso di sviluppo - vengono introdotti qui. I modelli rappresentano le due più comuni cause di stenosi dell&#39;arteria; cioè sono mostrati uno per ogni modello di topo iperplasia miointimale e aterosclerosi. Causare iperplasia miointimale, viene eseguita una lesione catetere a palloncino dell&#39;aorta addominale. Per lo sviluppo della placca aterosclerotica, l&#39;ApoE - è utilizzata modello del mouse in combinazione con dieta grassa occidentale - /. Diverse opzioni di modelli adattati per la misurazione e la valutazione dei risultati sono nominati e descritti in questo manoscritto. L&#39;introduzione e il confronto di questi due modelli fornisce information per gli scienziati di scegliere il modello di stenosi dell&#39;arteria adeguato in conformità alla questione scientifica posta.

La figura 1 mostra diverse opzioni di analisi e lo stato di malattia indotta per entrambi i modelli. Per il modello iperplasia intimale, viene mostrata l&#39;analisi di sezioni rappresentative trasversali in colorazione tricromica. Da queste sezioni trasversali, risultati come rapporti I / M, spessore massimo placca, spessore intimale massimo nel lume, obliterazione del lume in percentuale, così come l&#39;area della placca possono essere misurate e calcolate. Per valutare ...

Watch this Scientific Journal Video about Inducing Myointimal Hyperplasia Versus Atherosclerosis in Mice: An Introduction of Two Valid Models at JoVE.com

Inducing Myointimal Hyperplasia Versus Atherosclerosis in Mice: An Introduction of Two Valid Models

Questo video mostra due modelli di intimale sviluppo placca nelle arterie murini e sottolinea le differenze di iperplasia miointimale e aterosclerosi.

Indurre miointimale Iperplasia Versus aterosclerosi nei topi: Introduzione di due modelli validi

Various in vivo laboratory rodent models for the induction of artery stenosis have been established to mimic diseases that include arterial plaque ...

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12.0K Views.  University Hospital Hamburg.  Questo video mostra due modelli di intimale sviluppo placca nelle arterie murini e sottolinea le differenze di iperplasia miointimale e aterosclerosi. 

Video: Inducing Myointimal Hyperplasia Versus Atherosclerosis in Mice: An Introduction of Two Valid Models

Implantation of a Carotid Cuff for Triggering Shear-stress Induced Atherosclerosis in Mice

It is widely accepted that alterations in vascular shear stress trigger the expression of inflammatory genes in endothelial cells and thereby induce atherosclerosis (reviewed in 1 and 2). The role of shear stress has been extensively studied in vitro investigating the influence of flow dynamics on cultured endothelial cells 1,3,4 and in vivo in larger animals and humans 1,5,6,7,8. However, highly reproducible small animal models allowing systematic investigation of the influence of shear stress on plaque development are rare. Recently, Nam et al. 9 introduced a mouse model in which the ligation of branches of the carotid artery creates a region of low and oscillatory flow. Although this model causes endothelial dysfunction and rapid formation of atherosclerotic lesions in hyperlipidemic mice, it cannot be excluded that the observed inflammatory response is, at least in part, a consequence of endothelial and/or vessel damage due to ligation.
In order to avoid such limitations, a shear stress modifying cuff has been developed based upon calculated fluid dynamics, whose cone shaped inner lumen was selected to create defined regions of low, high and oscillatory shear stress within the common carotid artery 10. By applying this model in Apolipoprotein E (ApoE) knockout mice fed a high cholesterol western type diet, vascular lesions develop upstream and downstream from the cuff. Their phenotype is correlated with the regional flow dynamics 11 as confirmed by in vivo Magnetic Resonance Imaging (MRI) 12: Low and laminar shear stress upstream of the cuff causes the formation of extensive plaques of a more vulnerable phenotype, whereas oscillatory shear stress downstream of the cuff induces stable atherosclerotic lesions 11. In those regions of high shear stress and high laminar flow within the cuff, typically no atherosclerotic plaques are observed.
 
In conclusion, the shear stress-modifying cuff procedure is a reliable surgical approach to produce phenotypically different atherosclerotic lesions in ApoE-deficient mice.

The constricting cuff presented in this article is designed to induce atherosclerosis in the murine common carotid artery. Due to the conical shape of its inner lumen the implanted cuff generates well-defined regions of low, high and oscillatory shear stress triggering the development of atherosclerotic lesions of different inflammatory phenotypes.

Impianto di un bracciale carotidea per innescare Shear-stress aterosclerosi indotta in topi

It is widely accepted that alterations in vascular shear stress trigger the expression of inflammatory genes in endothelial cells and thereby induce ...

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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 ...

Minimal Invasive Surgical Procedure of Inducing Myocardial Infarction in Mice

Myocardial infarction still remains the main cause of death in western countries, despite considerable progress in the stent development area in the last decades. For clarification of the underlying mechanisms and the development of new therapeutic strategies, the availability of valid animal models are mandatory. Since we need new insights into pathomechanisms of cardiovascular diseases under in vivo conditions to combat myocardial infarction, the validity of the animal model is a crucial aspect. However, protection of animals are highly relevant in this context. Therefore, we establish a minimally invasive and simple model of myocardial infarction in mice, which assures a high reproducibility and survival rate of animals. Thus, this models fulfils the requirements of the 3R principle (Replacement, Refinement and Reduction) for animal experiments and assure the scientific information needed for further developing of therapeutical strategies for cardiovascular diseases.

A highly reproducible model for myocardial infarction in mice with minimal invasive manipulations is described. The model can be easily performed, resulting in a high reproducibility and survival rate. Thus, the described model will reduce the number of required animals as requested by the 3R principle (Replacement, Refinement and Reduction).

Minimal procedura chirurgica invasiva di indurre infarto miocardico nei topi

Myocardial infarction still remains the main cause of death in western countries, despite considerable progress in the stent development area in the last ...

Surgical Models of Gastroesophageal Reflux with Mice

Multiple surgical procedures have been reported to induce gastroesophageal reflux in animals. Herein, we report three surgical models with mice aiming to induce reflux of gastric contents, duodenal contents or mixed contents. Surgical procedures and general principles have been described in detail. A researcher with surgical experience should be able to grasp the technique after a short period of practice. After surgery, most mice can survive and develop reflux esophagitis similar to that in humans. However, it should be noted that histological differences between mouse and human esophagus are the inherent limitations of these surgical models. If used for research on Barrett&#8217;s esophagus and adenocarcinoma, these procedures may need to be combined with genetic modifications.

This article demonstrates surgical procedures of gastroesophageal reflux with mice. These models are useful tools for research on mechanisms and treatment of gastroesophageal reflux disease and potentially Barrett&#8217;s esophagus and esophageal adenocarcinoma.

I modelli chirurgici di reflusso gastroesofageo con Mice

Multiple surgical procedures have been reported to induce gastroesophageal reflux in animals. Herein, we report three surgical models with mice aiming to ...

An Improved Method for Rapid Intubation of the Trachea in Mice

Despite some anatomical and physiological differences, mouse models continue to be an essential tool for studying human lung disease. Bleomycin toxicity is a commonly used model to study both acute lung injury and fibrosis, and multiple methods have been developed for administering bleomycin (and other toxic agents) into the lungs. However, many of these approaches, such as transtracheal instillation, have inherent drawbacks, including the need for strong anesthetics and survival surgery. This paper reports a quick, reproducible method of intratracheal intubation that involves mild inhaled anesthesia, visualization of the trachea, and the use of a surrogate spirometer to confirm exposure. As a proof of concept, 8-12 week old C57BL/6 mice were administered either 2.0 U/kg of bleomycin or an equivalent volume of PBS, and both damage and fibrotic endpoints were measured post-exposure. This procedure allows researchers to treat a large cohort of mice in a relatively short period with little expense and minimal post-procedure care.

This article presents a rapid and simple method for administering bleomycin directly into the mouse trachea via intubation. Key advantages of this method are that it is highly reproducible, easy to master, and does not require specialized equipment or lengthy recovery times.

Un metodo migliore per Rapid intubazione della trachea nei topi

Despite some anatomical and physiological differences, mouse models continue to be an essential tool for studying human lung disease. Bleomycin toxicity ...

Induction of Accelerated Atherosclerosis in Mice: The "Wire-Injury" Model

Atherosclerosis is a proliferative fibro-inflammatory disease developing in the arterial wall, inducing a deficient blood flow or a lack of blood flow. Moreover, by rupture of the defective vascular wall, atherosclerosis induces occlusive thrombus formation, which represents the main cause of myocardial infarction or stroke and the most frequent cause of death. Despite the advances in the cardiovascular field, many questions remain unanswered, and additional basic research is essential to improve our understanding of the molecular mechanisms during atherosclerosis and its effects. Due to limited clinical studies, there is a need for representative animal models recreating atherosclerotic conditions such as neointima formation after stent implantation, balloon angioplasty, or endarterectomy. Since the mouse presents many advantages and is the most frequently used model for studying molecular processes, the current study proposes an invasive procedure of endothelial denudation, also known as the wire-injury model, which is representative of the human condition of neointima formation in arteries after revascularization procedures.

Induction of Accelerated Atherosclerosis in Mice: The &quot;Wire-Injury&quot; Model

This study describes an invasive procedure for the induction of accelerated atherosclerosis in mice. In comparison to other methods using electric- or cryo-induced injury, mechanical-induced injury mimics the human condition of restenosis after revascularization therapies and is ideal for the study of the molecular mechanisms involved.

Induzione di aterosclerosi accelerata nei topi: il modello "Wire-Injury"

Atherosclerosis is a proliferative fibro-inflammatory disease developing in the arterial wall, inducing a deficient blood flow or a lack of blood flow. ...

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 ...

Inducing Acute Lung Injury in Mice by Direct Intratracheal Lipopolysaccharide Instillation

Airway administration of lipopolysaccharide (LPS) is a common way to study pulmonary inflammation and acute lung injury (ALI) in small animal models. Various approaches have been described, such as the inhalation of aerosolized LPS as well as nasal or intratracheal instillation. The presented protocol describes a detailed step-by-step procedure to induce ALI in mice by direct intratracheal LPS instillation and perform FACS analysis of blood samples, bronchoalveolar lavage (BAL) fluid, and lung tissue. After intraperitoneal sedation, the trachea is exposed and LPS is administered via a 22 G venous catheter. A robust and reproducible inflammatory reaction with leukocyte invasion, upregulation of proinflammatory cytokines, and disruption of the alveolo-capillary barrier is induced within hours to days, depending on the LPS dosage used. Collection of blood samples, BAL fluid, and lung harvesting, as well as the processing for FACS analysis, are described in detail in the protocol. Although the use of the sterile LPS is not suitable to study pharmacologic interventions in infectious diseases, the described approach offers minimal invasiveness, simple handling, and good reproducibility to answer mechanistic immunological questions. Furthermore, dose titration as well as the use of alternative LPS preparations or mouse strains allow modulation of the clinical effects, which can exhibit different degrees of ALI severity or early vs. late onset of disease symptoms.

Presented here is a step-by-step procedure to induce acute lung injury in mice by direct intratracheal lipopolysaccharide instillation and to perform FACS analysis of blood samples, bronchoalveolar lavage fluid, and lung tissue. Minimal invasiveness, simple handling, good reproducibility, and titration of disease severity are advantages of this approach.

Indurre lesioni polmonari acute nei topi mediante instillazione diretta del lipopolioaccaride intracheale

Airway administration of lipopolysaccharide (LPS) is a common way to study pulmonary inflammation and acute lung injury (ALI) in small animal models. ...

A Revised Method for Inducing Secondary Lymphedema in the Hindlimb of Mice

Animal models are of paramount importance in the research of lymphedema in order to understand the pathophysiology of the disease but also to explore potential treatment options. This mouse model allows researchers to induce significant lymphedema lasting at least 8 weeks. Lymphedema is induced using a combination of fractioned radiotherapy and surgical ablation of lymphatics. This model requires that the mice get a dose of 10 Gray (Gy) radiation before and after surgery. The surgical part of the model involves ligation of three lymph vessels and extraction of two lymph nodes from the mouse hindlimb. Having access to microsurgical tools and a microscope is essential, due to the small anatomical structures of mice. The advantage of this model is that it results in statistically significant lymphedema, which provides a good basis for evaluating different treatment options. It is also a great and easily available option for microsurgical training. The limitation of this model is that the procedure can be time consuming, especially if not practiced in advance. The model results in objectively quantifiable lymphedema in mice, without causing severe morbidity and has been tested in three separate projects.

This animal model enables researchers to induce statistically significant secondary lymphedema in the hindlimb of mice, lasting at least 8 weeks. The model can be used to study the pathophysiology of lymphedema and to investigate novel treatment options.

Un metodo rivisto per indurre il linfomal secondario nell'Hindlimb dei topi

Animal models are of paramount importance in the research of lymphedema in order to understand the pathophysiology of the disease but also to explore ...

<meta charset="utf8"></head><body>Monitoraggio in tempo reale delle dinamiche cellulari nel trapianto di polmone di topo

Intravital Two-Photon Microscopy of the Transplanted Mouse Lung

Complications after lung transplantation are largely related to the host immune system responding to the graft. Such immune responses are regulated by crosstalk between donor and recipient cells. A better understanding of these processes relies on the use of preclinical animal models and is aided by an ability to study intra-graft immune cell trafficking in real-time. Intravital two-photon microscopy can be used to image tissues and organs for depths up to several hundred microns with minimal photodamage, which affords a great advantage over single-photon confocal microscopy. Selective use of transgenic mice with promoter-specific fluorescent protein expression and/or adoptive transfer of fluorescent dye-labeled cells during intravital two-photon microscopy allows for the dynamic study of single cells within their physiologic environment. Our group has developed a technique to stabilize mouse lungs, which has enabled us to image cellular dynamics in na&#239;ve lungs and orthotopically transplanted pulmonary grafts. This technique allows for detailed assessment of cellular behavior within the vasculature and in the interstitium, as well as for examination of interactions between various cell populations. This procedure can be readily learned and adapted to study immune mechanisms that regulate inflammatory and tolerogenic responses after lung transplantation. It can also be expanded to the study of other pathogenic pulmonary conditions.

<meta charset="utf8"></head><body>Autore in primo piano: Avanzamento dell'immunologia dei trapianti di polmone attraverso l'imaging intravitale

Here, we present a protocol to intravitally image the transplanted mouse left lung using two-photon microscopy. This represents a valuable tool for studying cellular dynamics and interactions in real-time following murine lung transplantation.

Microscopia intravitale a due fotoni del polmone di topo trapiantato

Complications after lung transplantation are largely related to the host immune system responding to the graft. Such immune responses are regulated by ...