Siamo grati per le risorse fornite dalla Harvard Medical School / Brigham and Women's Hospital Cardiovascular Physiology Core Laboratory. Questo lavoro è stato sostenuto in parte dai finanziamenti dei National Institutes of Health grants HL088533, HL071775, HL093148 e HL099073 (RL). MB ha ricevuto un premio di borsa di studio post-dottorato affiliato all'American Heart Association. KU è destinatario di un premio di borse di studio post-dottorato affiliato ai fondatori dell'American Heart Association. SC è stato supportato da un premio della Ellison Foundation.

1. Preparazione di immagini per le analisi
<ol><li>Ottenere immagini in modalità B ecocardiografica murina o umana nella vista parasternale ad asse lungo. Regola le impostazioni di compensazione del guadagno di tempo e il posizionamento della messa a fuoco di trasmissione per ottimizzare la visualizzazione dell'LV e di altre strutture cardiache nella vista parasternale, secondo la pratica abituale. Verificare che tutte le immagini vengano salvate in formato file DICOM. Le viste immagine standardizzate posizionano la parete ventricolare sinistra inferolaterale alla base del fotogramma. I telai devono mostrare l'intero miocardio ventricolare sinistro e il pericardio. La risoluzione deve essere abbastanza alta da delimitare il bordo pericardico, la parete del miocardio e il bordo endocardico del ventricolo sinistro. Scartare le immagini con un abbandono in eccesso o artefatti dell'immagine.</li>
	<li>Importare un file di immagine per l'analisi nella piattaforma software ImageJ v1.46 come file DICOM. Convertire il file in un file di immagine a 8 bit.</li>
	<li>Scorrere fotogrammi consecutivi del ciclo cardiaco fino a raggiungere un telaio end-diastolico di qualità appropriato. In alternativa, selezionare il frame end-diastolic in un programma di visualizzazione ecocardiografica e quindi esportare in un formato di file .jpg ad alta risoluzione da utilizzare in ImageJ. Identificare i fotogrammi più vicini alla diastole finale utilizzando l'onda R della traccia ECG, quindi identificare il singolo fotogramma migliore che cattura l'LV con la dimensione interna massima. Si consideri questo singolo fotogramma il telaio end-diastolic.</li>
	<li>Si suggerisce che gli utenti siano accecati dall'identità del soggetto quando selezionano le aree di interesse.</li>
</ol>2. Campionamento del ROI
<ol><li>Selezione dei riferimenti pericardiali. Quando si seleziona la regione di interesse pericardica (ROI), mirare a catturare l'eterogeneità del tessuto pericardico. Si noti che la luminosità e il contrasto dell'immagine possono essere regolati per la selezione del ROI, in base alle esigenze, senza alcun effetto sui risultati dell'analisi.<ol>
<li>Utilizzando lo strumento di disegno rettangolo di ImageJ, selezionate un rettangolo con lunghezza approssimante il terzo centrale della parete pericardico inferolaterale basale.</li>
		<li>Ridimensionare il ROI rettangolare per estenderlo sulla larghezza del pericardio utilizzando lo strumento di ridimensionamento del ROI.</li>
		<li>Ruotare il ROI in modo che si trova all'interno dell'area del pericardio utilizzando lo strumento di rotazione di ImageJ.</li>
		<li>Apportare le modifiche necessarie agli angoli del ROI pericardico. Cattura un'ultima regione di interesse pericardica che si trova all'interno del terzo centrale della parete pericardica, e include la larghezza della parete pericardica senza estendersi nelle regioni miocardiche o extra cardiache. Mirare a catturare la stessa posizione relativa e la percentuale dell'area totale del pericardio per tutte le misure adottate in un determinato studio.</li>
		<li>Applicare l'algoritmo alla selezione tramite gli strumenti di analisi ImageJ (vedere la sezione 3).</li>
	</ol>
</li>
	<li>Selezione del miocardio. Ancora una volta, mirare a catturare l'eterogeneità del tessuto miocardico all'interno del terzo centrale della parete miocardiale inferolaterale basale. Si noti che la luminosità e il contrasto dell'immagine possono essere regolati per la selezione del ROI, in base alle esigenze, senza alcun effetto sui risultati dell'analisi.<ol>
<li>Selezionare un rettangolo che si estende sulla larghezza della parete miocardico, escludendo l'endocardio e l'epicardio. Assicurarsi che la selezione del miocardio sia adiacente alla selezione del pericardio e allo stesso angolo theta. Non includere aree del muscolo papillare all'interno dell'area di selezione.</li>
		<li>Ruotare il ROI del miocardio in modo che si trovi parallelamente alla selezione del pericardio.</li>
		<li>Apportare le modifiche necessarie agli angoli del ROI del miocardio. Isolare un'ultima regione di interesse miocardica che si trova all'interno del terzo centrale della parete del miocardio e cattura la larghezza della parete senza estendersi nelle regioni pericardiche o intraluminali.</li>
		<li>Applicare l'algoritmo alla selezione tramite una macro ImageJ.</li>
	</ol>
</li>
</ol>3. Analisi ed elaborazione dei dati
<ol><li>Installare la macro ImageJ denominata "getHistogramValues.txt".</li>
	<li>Utilizzare lo strumento di analisi dell'istogramma ImageJ per visualizzare in anteprima la distribuzione dei valori di intensità del segnale all'interno del ROI (eseguire questo passaggio per la selezione del pericardico e per la selezione del miocardio).</li>
	<li>Utilizzare la macro ImageJ per registrare questi valori di densità del segnale per il ROI (eseguire questo passaggio per la selezione del pericardio e per la selezione del miocardio).<ol>
<li>Assegnare un valore di intensità da 0 (più scuro) a 255 unità (più luminose) a ogni pixel all'interno della selezione.</li>
		<li>Disporre i valori di intensità gerarchicamente, in ordine di intensità crescente, per produrre una distribuzione dell'intensità del segnale.</li>
		<li>Selezionare e segnalare i seguenti valori percentili per la distribuzione: 20° percentile, 50° percentile (mediana) e 80° percentile. </li>
	</ol>
</li>
	<li>Normalizzare le intensità miocardiali utilizzando il riferimento pericardico.<ol>
<li>Normalizzare dividendo i valori percentili miocardi di intensità per i corrispondenti valori percentili pericardiali di intensità12o sottraendo il valore percentile miocardico dell'intensità dal valore percentile pericardico di intensità13.</li>
		<li>Valori di report per i quattro metodi analitici: valori miocardio-pericardico normalizzati per i valori del 20° percentile, del 50° percentile (mediana) e dell'80° percentile. </li>
	</ol>
</li>
</ol>4. Quantificare la variabilità ciclica
<ol><li>Applicare l'algoritmo alle selezioni miocardico attraverso fotogrammi consecutivi del file DICOM, spostandosi attraverso il ciclo cardiaco. Confrontare le differenze nelle distribuzioni di intensità tra i fotogrammi, con attenzione ai fotogrammi end-sistolici e end-diastolic. Tutte le analisi delle immagini sopra descritte vengono eseguite offline su immagini ecocardiografiche nonive precedentemente acquisite e archiviate digitalmente in formato DICOM. Tutti i protocolli di studio sono stati approvati dal comitato di revisione istituzionale di Brigham and Women e dal Comitato istituzionale per la cura e l'uso degli animali dell'area medica di Harvard.</li>
</ol>

<ol>
	<li>Yamada, S., Komuro, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Integrated+backscatter+for+the+assessment+of+myocardial+viability.">Integrated backscatter for the assessment of myocardial viability.</a> Curr. Opin. Cardiol. 21, 433-437 (2006).</li><li>Mimbs, J. W., O'Donnell, M., Bauwens, D., Miller, J. W., Sobel, B. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+dependence+of+ultrasonic+attenuation+and+backscatter+on+collagen+content+in+dog+and+rabbit+hearts.">The dependence of ultrasonic attenuation and backscatter on collagen content in dog and rabbit hearts.</a> Circ. Res. 47, 49-58 (1980).</li><li>Picano, E., 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=In+vivo+quantitative+ultrasonic+evaluation+of+myocardial+fibrosis+in+humans.">In vivo quantitative ultrasonic evaluation of myocardial fibrosis in humans.</a> Circulation. 81, 58-64 (1990).</li><li>Mizuno, R., 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=Myocardial+ultrasonic+tissue+characterization+for+estimating+histological+abnormalities+in+hypertrophic+cardiomyopathy:+comparison+with+endomyocardial+biopsy+findings.">Myocardial ultrasonic tissue characterization for estimating histological abnormalities in hypertrophic cardiomyopathy: comparison with endomyocardial biopsy findings.</a> Cardiology. 96, 16-23 (2001).</li><li>Mizuno, R., Fujimoto, S., Saito, Y., Nakamura, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Non-invasive+quantitation+of+myocardial+fibrosis+using+combined+tissue+harmonic+imaging+and+integrated+backscatter+analysis+in+dilated+cardiomyopathy.">Non-invasive quantitation of myocardial fibrosis using combined tissue harmonic imaging and integrated backscatter analysis in dilated cardiomyopathy.</a> Cardiology. 108, 11-17 (2007).</li><li>Marini, 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=Cyclic+variation+in+myocardial+gray+level+as+a+marker+of+viability+in+man.+A+videodensitometric+study.">Cyclic variation in myocardial gray level as a marker of viability in man. A videodensitometric study.</a> Eur. Heart. J. 17, 472-479 (1996).</li><li>Komuro, K., 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=Sensitive+detection+of+myocardial+viability+in+chronic+coronary+artery+disease+by+ultrasonic+integrated+backscatter+analysis.">Sensitive detection of myocardial viability in chronic coronary artery disease by ultrasonic integrated backscatter analysis.</a> J. Am. Soc. Echocardiogr. 18, 26-31 (2005).</li><li>Ciulla, 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=Echocardiographic+patterns+of+myocardial+fibrosis+in+hypertensive+patients:+endomyocardial+biopsy+versus+ultrasonic+tissue+characterization.">Echocardiographic patterns of myocardial fibrosis in hypertensive patients: endomyocardial biopsy versus ultrasonic tissue characterization.</a> J. Am. Soc. Echocardiogr. 10, 657-664 (1997).</li><li>Maceira, A. M., Barba, J., Varo, N., Beloqui, O., Diez, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ultrasonic+backscatter+and+serum+marker+of+cardiac+fibrosis+in+hypertensives.">Ultrasonic backscatter and serum marker of cardiac fibrosis in hypertensives.</a> Hypertension. 39, 923-928 (2002).</li><li>D'Hooge, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=High+frame+rate+myocardial+integrated+backscatter.+Does+this+change+our+understanding+of+this+acoustic+parameter.">High frame rate myocardial integrated backscatter. Does this change our understanding of this acoustic parameter.</a> Eur. J. Echocardiogr. 1, 32-41 (2000).</li><li>Finch-Johnston, A. E., 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=Cyclic+variation+of+integrated+backscatter:+dependence+of+time+delay+on+the+echocardiographic+view+used+and+the+myocardial+segment+analyzed.">Cyclic variation of integrated backscatter: dependence of time delay on the echocardiographic view used and the myocardial segment analyzed.</a> J. Am. Soc. Echocardiogr. 13, 9-17 (2000).</li><li>Di Bello, V., 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=Increased+echodensity+of+myocardial+wall+in+the+diabetic+heart:+an+ultrasound+tissue+characterization+study.">Increased echodensity of myocardial wall in the diabetic heart: an ultrasound tissue characterization study.</a> J. Am. Coll. Cardiol. 25, 1408-1415 (1995).</li><li>Takiuchi, 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=Quantitative+ultrasonic+tissue+characterization+can+identify+high-risk+atherosclerotic+alteration+in+human+carotid+arteries.">Quantitative ultrasonic tissue characterization can identify high-risk atherosclerotic alteration in human carotid arteries.</a> Circulation. 102, 766-770 (2000).</li><li>Querejeta, R., 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=Serum+carboxy-terminal+propeptide+of+procollagen+type+I+is+a+marker+of+myocardial+fibrosis+in+hypertensive+heart+disease.">Serum carboxy-terminal propeptide of procollagen type I is a marker of myocardial fibrosis in hypertensive heart disease.</a> Circulation. 101, 1729-1735 (2000).</li></ol>

Nessun conflitto di interessi dichiarato.

Descriviamo il protocollo per un algoritmo di analisi delle immagini che quantifica la distribuzione dell'intensità del segnale sonografico e, a sua volta, offre una misura surrogata della microstruttura miocardica. Le funzionalità standardizzate del protocollo, tra cui la selezione, il dimensionamento e il posizionamento del ROI e dell'area di riferimento, servono a ridurre al minimo la variabilità basata su utenti e soggetti. Dimostriamo che se applicato alle immagini ecocardiografiche a fotogramma singolo end-diast...

L'ecocardiografia è una modalità di imaging ampiamente accessibile che è comunemente usata per caratterizzare e quantificare in modo non invasivo i cambiamenti nella struttura e nella funzione cardiaca. Le valutazioni ultrasoniche del tessuto cardiaco possono includere analisi dell'intensità del segnale di backscatter all'interno di una data regione di interesse in un singolo momento, nonché nel corso del ciclo cardiaco. Studi precedenti hanno suggerito che le misure di intensità del segnale sonografico possono identificare la presenza sottostante di disordine della fibra miocardica, tessuto miocardico vitale contro non vitale e fibrosi interstiziale1-3. Ci riferiamo alla 'microstruttura' miocardico come all'architettura tissutale che può essere caratterizzata, utilizzando l'analisi sonografica, al di là delle misurazioni lineari di dimensioni e morfologia lorde. Di conseguenza, le analisi dell'intensità del segnale sonografico sono state utilizzate per valutare le alterazioni microstrutturali del tessuto miocardico nell'impostazione della cardiomiopatia ipertrofica edilatata 4,5, della malattia coronarica cronica6,7e della cardiopatia ipertensiva8,9. Tuttavia, tecniche precedentemente stabilite si sono basate prevalentemente sul valore integrato o medio delle intensità del segnale backscatter, che possono essere suscettibili alla variabilità dal rumore casuale5, dati aliasati da bassi frame rates10e ritardi di tempo per gli algoritmi basati sulla variazione ciclica11.
Nel presente documento, descriviamo il metodo di utilizzo di un algoritmo di analisi delle immagini basato su ultrasuoni che si estende dai metodi precedenti; questo algoritmo si concentra su un singolo frame end-diastolic per l'analisi delle immagini e rappresenta la distribuzione completa dei valori di intensità del segnale derivati da un dato campione miocardico. Utilizzando il pericardio come riferimento in-frame12,13, l'algoritmo quantifica in modo riproducibile la variazione nelle distribuzioni dell'intensità del segnale sonografico e offre una misura surrogata migliorata della microstruttura miocardica. In un protocollo passo-passo, descriviamo i metodi per preparare le immagini per l'uso, campio campiodando le aree di interesse e elaborando i dati all'interno di aree di interesse selezionate. Mostriamo anche risultati rappresentativi dall'applicazione dell'algoritmo alle immagini ecocardiografiche acquisite da topi e umani con esposizione variabile allo stress post-carico sul ventricolo sinistro.

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		<tr height="21">
			<td height="21" style="height:21px;width:216px;">ImageJ v 1.46</td>
			<td style="width:268px;">NIH (Bethesda, MD)</td>
			<td style="width:119px;"></td>
			<td style="width:167px;">open access software</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Power ShowCase</td>
			<td style="width:268px;">Trillium Technology (Ann Arbor, MI)</td>
			<td style="width:119px;"></td>
			<td>commercial software</td>
		</tr>
	</tbody>
</table>

ultrasonic assessment of myocardial microstructure

L'ecocardiografia è una modalità di imaging ampiamente accessibile che è comunemente usata per caratterizzare e quantificare in modo non invasivo i cambiamenti nella struttura e nella funzione cardiaca. Le valutazioni ultrasoniche del tessuto cardiaco possono includere analisi dell'intensità del segnale di backscatter all'interno di una data regione di interesse. Le tecniche precedentemente stabilite si sono basate prevalentemente sul valore integrato o medio delle intensità del segnale backscatter, che possono essere suscettibili alla variabilità dai dati alias da bassi frame rates e ritardi di tempo per gli algoritmi basati sulla variazione ciclica. Nel presente documento, descriviamo un algoritmo di imaging basato su ultrasuoni che si estende dai metodi precedenti, può essere applicato a un singolo fotogramma dell'immagine e rappresenta la distribuzione completa dei valori di intensità del segnale derivati da un dato campione di miocardio. Se applicato a dati rappresentativi di imaging del mouse e dell'uomo, l'algoritmo distingue tra soggetti con e senza esposizione alla resistenza cronica al carico post-carico. L'algoritmo offre una misura surrogata migliorata della microstruttura miocardico e può essere eseguito utilizzando un software di analisi delle immagini ad accesso aperto.

L'analisi dell'intensità del segnale viene eseguita in 4 passaggi principali(Figura 1),tra cui: 1) selezione e formattazione dell'immagine, 2) ROI di campionamento e aree di riferimento, 3) applicazione dell'algoritmo e 4) elaborazione dei valori finali per ottenere rapporti di intensità miocardica-pericardica. La selezione e le dimensioni del ROI sono standardizzate per limitare la variabilità tra utenti e intrautente (Figura 2). Il posizionamento di ogni ROI è anche standardizzato ...

Watch this Scientific Journal Video about Ultrasonic Assessment of Myocardial Microstructure at JoVE.com

Ultrasonic Assessment of Myocardial Microstructure

L'ecocardiografia è comunemente usata per caratterizzare e quantificare in modo non invasivo i cambiamenti nella struttura e nella funzione cardiaca. Descriviamo un algoritmo di imaging basato su ultrasuoni che offre una misura surrogata migliorata della microstruttura miocardico e può essere eseguito utilizzando un software di analisi delle immagini ad accesso aperto.

Valutazione ad ultrasuoni della microstruttura miocardica

Echocardiography is a widely accessible imaging modality that is commonly used to noninvasively characterize and quantify changes in cardiac structure and ...

ultrasonic-assessment-of-myocardial-microstructure

Research

JoVE Journal

Medicine

5.4K Views.  Harvard Medical School. L'ecocardiografia è comunemente usata per caratterizzare e quantificare in modo non invasivo i cambiamenti nella struttura e nella funzione cardiaca. Descriviamo un algoritmo di imaging basato su ultrasuoni che offre una misura surrogata migliorata della microstruttura miocardico e può essere eseguito utilizzando un software di analisi delle immagini ad accesso aperto.

Video: Ultrasonic Assessment of Myocardial Microstructure

Magnetic Resonance Derived Myocardial Strain Assessment Using Feature Tracking

Purpose: An accurate and practical method to measure parameters like strain in myocardial tissue is of great clinical value, since it has been shown, that strain is a more sensitive and earlier marker for contractile dysfunction than the frequently used parameter EF. Current technologies for CMR are time consuming and difficult to implement in clinical practice. Feature tracking is a technology that can lead to more automization and robustness of quantitative analysis of medical images with less time consumption than comparable methods.
Methods: An automatic or manual input in a single phase serves as an initialization from which the system starts to track the displacement of individual patterns representing anatomical structures over time. The specialty of this method is that the images do not need to be manipulated in any way beforehand like e.g. tagging of CMR images.
Results: The method is very well suited for tracking muscular tissue and with this allowing quantitative elaboration of myocardium and also blood flow.
Conclusions: This new method offers a robust and time saving procedure to quantify myocardial tissue and blood with displacement, velocity and deformation parameters on regular sequences of CMR imaging. It therefore can be implemented in clinical practice.

An accurate and practical method to measure parameters like strain in myocardial tissue is of great clinical value, since it has been shown, that strain is a more sensitive and earlier marker for contractile dysfunction than the frequently used parameter EF.

Risonanza Magnetica Derivato valutazione di deformazione miocardica Utilizzo del rilevamento delle caratteristiche

Purpose: An accurate and practical method to measure parameters like strain in myocardial tissue is of great clinical value, since it has been shown, that ...

Ricerca

Medicina

Acute Myocardial Infarction in Rats

With heart failure leading the cause of death in the USA (Hunt), biomedical research is fundamental to advance medical treatments for cardiovascular diseases. Animal models that mimic human cardiac disease, such as myocardial infarction (MI) and ischemia-reperfusion (IR) that induces heart failure as well as pressure-overload (transverse aortic constriction) that induces cardiac hypertrophy and heart failure (Goldman and Tarnavski), are useful models to study cardiovascular disease. In particular, myocardial ischemia (MI) is a leading cause for cardiovascular morbidity and mortality despite controlling certain risk factors such as arteriosclerosis and treatments via surgical intervention (Thygesen). Furthermore, an acute loss of the myocardium following myocardial ischemia (MI) results in increased loading conditions that induces ventricular remodeling of the infarcted border zone and the remote non-infarcted myocardium. Myocyte apoptosis, necrosis and the resultant increased hemodynamic load activate multiple biochemical intracellular signaling that initiates LV dilatation, hypertrophy, ventricular shape distortion, and collagen scar formation. This pathological remodeling and failure to normalize the increased wall stresses results in progressive dilatation, recruitment of the border zone myocardium into the scar, and eventually deterioration in myocardial contractile function (i.e. heart failure). The progression of LV dysfunction and heart failure in rats is similar to that observed in patients who sustain a large myocardial infarction, survive and subsequently develops heart failure (Goldman). The acute myocardial infarction (AMI) model in rats has been used to mimic human cardiovascular disease; specifically used to study cardiac signaling mechanisms associated with heart failure as well as to assess the contribution of therapeutic strategies for the treatment of heart failure. The method described in this report is the rat model of acute myocardial infarction (AMI). This model is also referred to as an acute ischemic cardiomyopathy or ischemia followed by reperfusion (IR); which is induced by an acute 30-minute period of ischemia by ligation of the left anterior descending artery (LAD) followed by reperfusion of the tissue by releasing the LAD ligation (Vasilyev and McConnell). This protocol will focus on assessment of the infarct size and the area-at-risk (AAR) by Evan's blue dye and triphenyl tetrazolium chloride (TTC) following 4-hours of reperfusion; additional comments toward the evaluation of cardiac function and remodeling by modifying the duration of reperfusion, is also presented. Overall, this AMI rat animal model is useful for studying the consequence of a myocardial infarction on cardiac pathophysiological and physiological function.

The rat model of acute myocardial infarction (AMI) is useful to study the consequence of a MI on cardiac pathophysiological and physiological function.

Infarto acuto del miocardio in topi

With heart failure leading the cause of death in the USA (Hunt), biomedical research is fundamental to advance medical treatments for cardiovascular ...

Assessment of Cardiac Function and Myocardial Morphology Using Small Animal Look-locker Inversion Recovery (SALLI) MRI in Rats

Small animal magnetic resonance imaging is an important tool to study cardiac function and changes in myocardial tissue. The high heart rates of small animals (200 to 600 beats/min) have previously limited the role of CMR imaging. Small animal Look-Locker inversion recovery (SALLI) is a T1 mapping sequence for small animals to overcome this problem 1. T1 maps provide quantitative information about tissue alterations and contrast agent kinetics. It is also possible to detect diffuse myocardial processes such as interstitial fibrosis or edema 1-6. Furthermore, from a single set of image data, it is possible to examine heart function and myocardial scarring by generating cine and inversion recovery-prepared late gadolinium enhancement-type MR images 1.
	The presented video shows step-by-step the procedures to perform small animal CMR imaging. Here it is presented with a healthy Sprague-Dawley rat, however naturally it can be extended to different cardiac small animal models.

Assessment of Cardiac Function and Myocardial Morphology Using Small Animal Look-locker Inversion Recovery SALLI MRI in Rats

Contrast enhanced cardiac magnetic resonance (CMR) imaging allows comprehensive in vivo assessment of the heart in small animal models of cardiovascular disease. Here we detail the procedures to perform CMR imaging, reconstruction and analysis using Small Animal Lock-Locker Inversion Recovery (SALLI) in rats.

Valutazione della funzione cardiaca e Morfologia miocardica Utilizzando Small Animal Look-Locker Inversion Recovery (SALLI) MRI nei ratti

Small animal magnetic resonance  imaging is an important tool to study cardiac function and changes in myocardial  tissue. The high heart rates of small ...

Myocardial Infarction and Functional Outcome Assessment in Pigs

Introduction of newly discovered cardiovascular therapeutics into first-in-man trials depends on a strictly regulated ethical and legal roadmap. One important prerequisite is a good understanding of all safety and efficacy aspects obtained in a large animal model that validly reflect the human scenario of myocardial infarction (MI). Pigs are widely used in this regard since their cardiac size, hemodynamics, and coronary anatomy are close to that of humans. Here, we present an effective protocol for using the porcine MI model using a closed-chest coronary balloon occlusion of the left anterior descending artery (LAD), followed by reperfusion. This approach is based on 90 min of myocardial ischemia, inducing large left ventricle infarction of the anterior, septal and inferoseptal walls. Furthermore, we present protocols for various measures of outcome that provide a wide range of information on the heart, such as cardiac systolic and diastolic function, hemodynamics, coronary flow velocity, microvascular resistance, and infarct size. This protocol can be easily tailored to meet study specific requirements for the validation of novel cardioregenerative biologics at different stages (i.e. directly after the acute ischemic insult, in the subacute setting or even in the chronic MI once scar formation has been completed). This model therefore provides a useful translational tool to study MI, subsequent adverse remodeling, and the potential of novel cardioregenerative agents.

This protocol describes the porcine myocardial infarction (MI) model using a 90 min closed-chest coronary balloon occlusion of the left anterior descending artery (LAD), followed by reperfusion. Furthermore, the protocol for several outcome parameters, such as cardiac function, hemodynamics, microvascular resistance, and infarct size, are also presented.

Infarto miocardico e funzionale Outcome Assessment in Pigs

Introduction of newly discovered cardiovascular therapeutics into first-in-man trials depends on a strictly regulated ethical and legal roadmap. One ...

Primary Outcome Assessment in a Pig Model of Acute Myocardial Infarction

Mortality after acute myocardial infarction remains substantial and is associated with significant morbidity, like heart failure. Novel therapeutics are therefore required to confine cardiac damage, promote survival and reduce the disease burden of heart failure. Large animal experiments are an essential part in the translational process from experimental to clinical therapies. To optimize clinical translation, robust and representative outcome measures are mandatory. The present manuscript aims to address this need by describing the assessment of three clinically relevant outcome modalities in a pig acute myocardial infarction (AMI) model: infarct size in relation to area at risk (IS/AAR) staining, 3-dimensional transesophageal echocardiography (TEE) and admittance-based pressure-volume (PV) loops. Infarct size is the main determinant driving the transition from AMI to heart failure and can be quantified by IS/AAR staining. Echocardiography is a reliable and robust tool in the assessment of global and regional cardiac function in clinical cardiology. Here, a method for three-dimensional transesophageal echocardiography (3D-TEE) in pigs is provided. Extensive insight into cardiac performance can be obtained by admittance-based pressure-volume (PV) loops, including intrinsic parameters of myocardial function that are pre- and afterload independent. Combined with a clinically feasible experimental study protocol, these outcome measures provide researchers with essential information to determine whether novel therapeutic strategies could yield promising targets for future testing in clinical studies.

Reliable and accurate outcome assessment is the key for translation of preclinical therapies into clinical treatment. The current paper describes how to assess three clinically relevant primary outcome parameters of cardiac performance and damage in a pig acute myocardial infarction model.

La valutazione dell&#39;outcome primario in un modello suino di infarto miocardico acuto

Mortality after acute myocardial infarction remains substantial and is associated with significant morbidity, like heart failure. Novel therapeutics are ...

Transthoracic Speckle Tracking Echocardiography for the Quantitative Assessment of Left Ventricular Myocardial Deformation

The value of conventional echocardiography is limited by differences in inter-individual image interpretation and therefore largely dependent on the examiners&#39; expertise. Speckle tracking Echocardiography (STE) is a promising but technically challenging method that can be used to quantitatively assess regional and global systolic and diastolic myocardial performance. Myocardial strain and strain rate can be measured in all three dimensions&#160;&#8212; radial, circumferential, longitudinal &#8212; of myocardial deformation. Standard cross-sectional two-dimensional B-mode images are recorded and subsequently postprocessed by automated continuous frame-by-frame tracking and motion analysis of speckles within the myocardium. Images are recorded as digital loops and synchronized to a 3-lead EKG for timing purposes. Longitudinal deformation is assessed in the apical 4-, 3-, and 2-chamber views. Circumferential and radial deformation are measured in the parasternal short axis plane.
Optimal image quality and accurate tissue tracking are paramount for the correct determination of myocardial performance parameters. Utilizing transthoracic STE in a healthy volunteer, the present article is a detailed outline of the essential steps and potential pitfalls of quantitative echocardiographic myocardial deformation analysis.

Speckle tracking echocardiography is an emerging diagnostic imaging technique for the quantitative assessment of global and regional myocardial performance. Standard view echocardiographic motion images are recorded and deformation parameters are subsequently measured by automated continuous frame-by-frame tracking and motion analysis of speckles within the B-mode images of the myocardium.

Transtoracica Speckle monitoraggio ecocardiografia per la valutazione quantitativa del ventricolo sinistro del miocardio Deformazione

The value of conventional echocardiography is limited by differences in inter-individual image interpretation and therefore largely dependent on the ...

Histological Quantification of Chronic Myocardial Infarct in Rats

Myocardial infarction is defined as cardiomyocyte death due to prolonged ischemia; an inflammatory response and scar formation (fibrosis) follow the ischemic injury. Following the initial acute phase, chronic remodeling of the left ventricle (LV) modifies the structure and function of the heart. Permanent coronary ligation in small animals has been widely used as a reference model for a chronic model of MI. Thinning of the infarcted wall progressively develops to transmural fibrosis. Histological assessment of infarct size is commonly performed; nevertheless, a standardization of the methods for quantification is missing. Indeed, important methodological aspects, such as the number of sections analyzed and the sampling and quantification methods, are usually not described and therefore preclude comparison across investigations. Too often, quantification is performed on a single section obtained at the level of the papillary muscles. Because novel strategies aimed at reducing infarct expansion and remodeling are under investigation, there is an important need for the standardization of accurate heart sampling protocols. We describe an accurate method to quantify the infarct size using a systematic sampling of harvested rat heart and image analyses of trichromatic stained histological sections obtained from base to apex. We also provide evidence that calculating the expansion index (EI) allowed for infarct size assessment, taking into account changes of the left ventricle throughout the remodeling.

The post-mortem assessment of myocardial infarction (MI) in rodents is based on quantification of the infarct on stained heart sections. We describe an accurate method to quantify the infarct size using systematic sampling of harvested rat hearts from base to apex and image analyses of trichrome-stained histological sections.

Quantificazione istologica di cronica Myocardial Infarto in Rats

Myocardial infarction is defined as cardiomyocyte death due to prolonged ischemia; an inflammatory response and scar formation (fibrosis) follow the ...

3D Whole-heart Myocardial Tissue Analysis

Cardiac regenerative therapies aim to protect and repair the injured heart in patients with ischemic heart disease. By injecting stem cells or other biologicals that enhance angio- or vasculogenesis into the infarct border zone (IBZ), tissue perfusion is improved, and the myocardium can be protected from further damage. For maximum therapeutic effect, it is hypothesized that the regenerative substance is best delivered to the IBZ. This requires accurate injections and has led to the development of new injection techniques. To validate these new techniques, we have designed a validation protocol based on myocardial tissue analysis. This protocol includes whole-heart myocardial tissue processing that enables detailed two-dimensional (2D) and three-dimensional (3D) analysis of the cardiac anatomy and intramyocardial injections. In a pig, myocardial infarction was created by a 90-min occlusion of the left anterior descending coronary artery. Four weeks later, a mixture of a hydrogel with superparamagnetic iron oxide particles (SPIOs) and fluorescent beads was injected in the IBZ using a minimally-invasive endocardial approach. 1 h after the injection procedure, the pig was euthanized, and the heart was excised and embedded in agarose (agar). After the solidification of the agar, magnetic resonance imaging (MRI), slicing of the heart, and fluorescence imaging were performed. After image post-processing, 3D analysis was performed to assess the IBZ targeting accuracy. This protocol provides a structured and reproducible method for the assessment of the targeting accuracy of intramyocardial injections into the IBZ. The protocol can be easily used when the processing of scar tissue and/or validation of the injection accuracy of the whole heart is desired.

This protocol describes a novel method for the 3D comparison of whole-heart myocardial tissue with MRI. This is designed for the accurate assessment of intramyocardial injections in the infarct border zone of a chronic porcine model of myocardial infarction.

3D Whole-cuore Analisi tessuto miocardico

Cardiac regenerative therapies aim to protect and repair the injured heart in patients with ischemic heart disease. By injecting stem cells or other ...

Ultrasonic-augmented Primary Adult Fibroblast Isolation

Primary adult fibroblasts have become an important tool to study fibrosis, fibroblast interactions and inflammation in all body tissues. Since primary fibroblasts cannot divide indefinitely due to myofibroblast differentiation or senescence induction, new cultures must be established regularly. However, there are several obstacles to overcome during the processes of developing a reliable isolation protocol and primary fibroblast isolation itself: the method&#8217;s degree of difficulty (especially for beginners), the risk of bacterial contamination, the required time until primary fibroblasts can be used for experiments, and subsequent cell quality and viability. In this study, a fast, reliable and easy-to-learn protocol to isolate and culture primary adult fibroblasts from mouse heart, lung, liver and kidney combining enzymatic digestion and ultrasonic agitation is provided.

We present a protocol to isolate primary adult fibroblasts in an easy, fast and reliable way, performable by beginners (e.g., students). The procedure combines enzymatic tissue digestion and mechanical agitation with ultrasonic waves to obtain primary fibroblasts. The protocol can easily be adapted to specific experimental requirements (e.g., human tissue).

Isolamento fibroblasto adulto primario aumentata ad ultrasuoni

Primary adult fibroblasts have become an important tool to study fibrosis, fibroblast interactions and inflammation in all body tissues. Since primary ...

Semi-Minimal Invasive Method to Induce Myocardial Infarction in Rats and the Assessment of Cardiac Function by an Isolated Working Heart System

Myocardial infarction (MI) remains the main contributor to morbidity and mortality worldwide. Therefore, research on this topic is mandatory. An easily and highly reproducible MI induction procedure is required to obtain further insight and better understanding of the underlying pathological changes. This procedure can also be used to evaluate the effects or potency of new and promising treatments (as drugs or interventions) in acute MI, subsequent remodeling and heart failure (HF). After intubation and pre-operative preparation of the animal, an anesthetic protocol with isoflurane was performed, and the surgical procedure was conducted&#160;quickly. Using a minimally invasive approach, the left anterior descending artery (LAD) was located and occluded by a ligature. The occlusion can be performed acutely for subsequent reperfusion (ischemia/reperfusion injury). Alternatively, the vessel can be ligated permanently to investigate the development of chronic MI, remodeling or HF. Despite common pitfalls, the drop-out rates are minimal. Various treatments such as remote ischemic conditioning can be examined for their cardioprotective potential pre-, peri- and post-operatively. The post-operative recovery was quick as the anesthesia was precisely controlled and the duration of the operation was short. Post-operative analgesia was administered for three days. The minimally invasive procedure reduces the risk of infection and inflammation. Furthermore, it facilitates rapid recovery. The &#8220;working heart&#8221; measurements were performed ex vivo and enabled precise control of preload, afterload and flow. This procedure requires specific equipment and training for adequate performance. This manuscript provides a detailed step-by-step introduction for conducting these measurements.

This article presents an efficient method to perform myocardial ischemia and subsequent chronic reperfusion in rats using a minimally invasive approach. In addition, left ventricular hemodynamic function of rats is assessed by echocardiography and isolated working heart methods.&#160;

Metodo semi-minimo invasivo per indurre l'infarto miocardico nei ratti e la valutazione della funzione cardiaca da parte di un sistema cardiaco funzionante isolato

Myocardial infarction (MI) remains the main contributor to morbidity and mortality worldwide. Therefore, research on this topic is mandatory. An easily ...