Projeto nº. NVKP_16-1-2016-0017 («Programa Nacional do Coração») foi implementado com o apoio do Fundo Nacional de Investigação, Desenvolvimento e Inovação da Hungria, financiado ao abrigo do regime de financiamento NVKP_16. A investigação foi financiada pelo Programa Temático de Excelência (2020-4.1.1.-TKP2020) do Ministério da Inovação e Tecnologia da Hungria, no âmbito dos programas temáticos de Desenvolvimento Terapêutico e Bioimagem da Universidade Semmelweis.

O protocolo segue as diretrizes do comitê de ética em pesquisa com seres humanos da instituição e os pacientes dos casos clínicos assinaram o termo de consentimento livre e esclarecido para o estudo.
1. Requisitos técnicos
<ol><li>Para aquisição e análise 3D, use software e hardware apropriados. Use cabos de ECG do aparelho de ecocardiografia; além disso, é obrigatório para o protocolo completo de aquisição 3D descrito abaixo.</li>
	<li>Para aquisição 3D, use uma sonda ecocardiográfica 3D e uma máquina de ultrassom compatível com 3D. Para análise volumétrica de RV 3D, use um software dedicado.</li>
</ol>2. Aquisição
<ol><li>Na grande maioria dos casos, realizar a aquisição 3D do VD utilizando vistas apicais. Em oposição às visões focadas no VE, recomenda-se um posicionamento diferente do paciente. Se uma qualidade de imagem substancialmente melhor puder ser alcançada alternando para um espaço intercostal sobre a visão apical correta, essa visão encurtada pode permitir uma melhor qualidade de imagem 3D. O encurtamento pode ser corrigido durante a análise 3D.<ol><li>Em comparação com a aquisição ecocardiográfica apical padrão, onde a posição de decúbito lateral esquerdo (paciente deitado do lado esquerdo com o braço esquerdo esticado acima da cabeça) é recomendada, faça com que o paciente se incline um pouco mais para trás para permitir uma posição mais lateral do transdutor.</li>
		<li>Escolha uma profundidade de imagem que inclua apenas o RV. A profundidade desnecessariamente grande pode diminuir a taxa de quadros de aquisição com a falta de efeitos benéficos em relação à análise volumétrica do RV.</li>
	</ol></li>
	<li>Confirme a visão correta focada no RV a partir de imagens de ecocardiografia 2D. Se a parede livre do RV for mal visualizada, mesmo a partir dessa visão, a qualidade de imagem 3D esperada não será ideal para análises posteriores.</li>
	<li>Mude para a imagem 3D ao vivo usando o botão 4D , onde a correção adicional da visão RV pode ser realizada.</li>
	<li>Embora o modo ao vivo 3D possa ser bastante agradável esteticamente, use o modo 12 Slice para a visualização 3D, que mostra uma imagem triplana da região de interesse, bem como 9 planos transversais que podem ser modificados livremente. Por rotação e posicionamento correto dos planos de corte, confirme a visibilidade de toda a parede livre de RV (incluindo a faixa de saída e os segmentos apicais).</li>
	<li>Ajuste ainda mais a imagem usando a inclinação esquerda do setor (segunda página na tela sensível ao toque) para melhorar a visualização do RV.</li>
	<li>Use dois modos de aquisição 3D para análise volumétrica RV: o modo multi beat e o modo single beat . Use ambas as abordagens em todos os pacientes, no entanto, em alguns casos (por exemplo, certas arritmias, dispneia grave do paciente), apenas a última pode ser viável.</li>
	<li>Usando o modo de batida única , obtenha uma troca entre a qualidade da imagem e a taxa de quadros. Escolha uma profundidade, largura e taxa de quadros de imagem ideais (painel inferior da tela sensível ao toque) e obtenha loops 3D do RV sem qualquer ação adicional. Este método é viável na maioria dos pacientes; no entanto, ele produz geralmente menor qualidade de imagem e taxa de quadros em comparação com a abordagem multi beat .<ol><li>No caso de uma frequência cardíaca média (60-70/min), mantenha um limite de frequência de quadros mais baixo de 16 quadros/s para uma análise adequada do VD; no entanto, se a taquicardia estiver presente, recomendam-se taxas de quadros ainda mais altas.</li>
	</ol></li>
	<li>Usando o modo multi beat , reconstrua o loop 3D adquirido a partir de um determinado número de ciclos cardíacos que podem ser selecionados na tela sensível ao toque (os modos de 2,3,4 e 6 batimentos podem ser usados). Em contraste com a aquisição de batida única, geralmente espera-se melhor qualidade de imagem e taxa de quadros; no entanto, requer comprimentos de ciclo cardíaco relativamente constantes e também a adesão do paciente devido à manobra obrigatória de apneia. A manobra é essencial para evitar os chamados artefatos de costura: quando o volume 3D adquirido é costurado, comprimentos de ciclo cardíaco e/ou movimento desiguais devido à respiração podem resultar nesse fenômeno.<ol><li>Após o posicionamento correto da sonda e a configuração da máquina (semelhante ao modo de "batida única"), peça ao paciente para respirar fundo e segurá-la. Neste caso, os pulmões em expansão geralmente cobrem toda a imagem.</li>
		<li>Peça ao paciente para expirar lentamente, estritamente com orientação. Em paralelo com a deflação dos pulmões, o VD torna-se visível novamente.</li>
		<li>Quando todo o VD (parede livre e septo) reaparecer, peça ao paciente para prender a respiração neste estado.</li>
		<li>Ao clicar em multi batida na tela, inicie a aquisição e o loop 3D se acumula durante a quantidade determinada de ciclos cardíacos.</li>
		<li>Quando a aquisição estiver pronta (todo o VD é visualizado), peça ao paciente para respirar livremente novamente.</li>
		<li>Verifique o loop obtido para garantir se não há artefatos de costura ou desistência.</li>
	</ol></li>
</ol>3. Análise 4D RV
<ol><li>Usando um software dedicado, realize análises volumétricas 3D do RV. Depois de escolher o loop 3D focado em RV na biblioteca do paciente, abra o software na janela Medição encontrada na pasta Volume .</li>
	<li>Depois de abrir o software, oriente o RV em quatro planos de corte predefinidos.<ol><li>Coloque dois marcadores (TV Center) no centro da válvula tricúspide nos planos superior e inferior esquerdo de eixo longo. Ajuste o eixo longo da imagem para o eixo longo real do RV usando a ferramenta de rotação. As imagens de referência nas bordas superiores direitas mostram como a orientação correta deve aparecer.</li>
		<li>Nos painéis superior e inferior direito, alinhe as imagens de eixo curto na posição correta por rotação. Da mesma forma que a etapa anterior, as imagens de referência também ajudam nesse processo.</li>
	</ol></li>
	<li>Depois de concluir, clique em Definir pontos de referência para a próxima etapa da análise. Defina pontos de referência em duas imagens.<ol><li>No lado esquerdo, marque o anel tricúspide na parede livre (parede livre de TV) e septo (septo de TV) e o ápice do VD na visão apical de quatro câmaras previamente orientada.</li>
		<li>No lado direito, coloque-se os pontos de inserção posterior do VD (VE/VD posterior) e anterior (VE/VD anterior) e a parede livre do VD (parede livre do VD). Da mesma forma que a janela anterior, as imagens de referência no canto superior direito ajudam a verificar a configuração correta. Depois de definir todos os pontos de referência, o software salta automaticamente para a próxima janela (Revisão).</li>
	</ol></li>
	<li>Nesta janela (Revisão), revise e corrija manualmente a detecção automática da borda endocárdica ao longo de todo o ciclo cardíaco, se necessário. Por padrão, 9 painéis podem ser vistos: no lado esquerdo, 3 loops móveis (1 eixo longo e 2 eixo curto), no meio os quadros diastólicos finais das mesmas imagens e no lado direito os sistólicos finais.<ol><li>No caso de rastreamento falso, corrija livremente as bordas endocárdicas (linhas verdes), a borda rastreada clicando nelas. Usando a ferramenta de rotação no eixo curto, revise o rastreamento de imagens ao longo de toda a circunferência do RV. Ajuste a magnitude da correção escolhendo o tamanho da caneta no painel do lado direito. Se o rastreamento for considerado correto, clique em Resultados no mesmo painel.</li>
	</ol></li>
	<li>Na última seção, revise os dados volumétricos 3D finais e outros parâmetros calculados no lado superior direito (painel Planilha ). Além dos volumes RV e da fração de ejeção, o software também exibe parâmetros 2D, como diâmetros lineares (médios, basais e de eixo longo), bem como valores FAC e TAPSE derivados da visualização apical de quatro câmaras predefinida. O software também mostra um eixo longo e curto do RV (lado esquerdo), um modelo 3D ao vivo do RV (meio superior) e uma curva volume-tempo da câmara (inferior direita).<ol><li>Em caso de necessidade de mais ajustes no rastreamento, todas as etapas anteriores estão disponíveis para correção, clicando nelas no painel direito. Se o rastreamento e os parâmetros 3D forem considerados válidos, salve os resultados clicando em "Aprovar e sair" no mesmo painel.</li>
	</ol></li>
</ol>

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A análise 3D do VD representa um passo importante na prática cardiológica cotidiana. Paralelamente ao crescente interesse da morfologia e função da câmara cardíaca anteriormente negligenciada, essas novas soluções fornecem informações clinicamente significativas sobre o lado direito do coração. Embora a aquisição 3D tenha vários aspectos que diferem marcadamente da imagem ecocardiográfica 2D, mantendo atenção especial aos pontos críticos e usando um protocolo completo, a análise 3D do VD pode progred...

A ecocardiografia percorreu um longo caminho desde suas primeiras aplicações clínicas na década de 19501. As primeiras sondas de ultrassom unidimensionais foram projetadas para fornecer diâmetros lineares simples das paredes da câmara e lúmens; no entanto, eles representam, sem dúvida, um marco na imagem cardiovascular. O desenvolvimento da ultrassonografia bidimensional (2D) foi outro passo importante, fornecendo uma quantificação muito mais precisa da morfologia e da função, e ainda é considerado o método padrão na prática clínica cotidiana. No entanto, a avaliação baseada em ecocardiografia 2D ainda carrega uma grande limitação da técnica: a imagem de uma determinada câmara a partir de alguns planos tomográficos não caracteriza adequadamente a morfologia e a função de uma estrutura tridimensional (3D). Esse problema é ainda mais pronunciado no caso do ventrículo direito (VD): em comparação com o ventrículo esquerdo (VE) relativamente simples, em forma de bala, o VD possui uma geometria complexa2 que não pode ser adequadamente quantificada usando diâmetros lineares ou áreas3. Apesar desses fatos amplamente conhecidos, a morfologia e a função do VD geralmente são medidas por parâmetros tão simples na prática clínica.
Por muitas décadas, o RV foi considerado como tendo um papel muito menos importante na circulação em comparação com sua contraparte esquerda. Vários artigos de referência derrotaram esse ponto de vista, mostrando o forte papel prognóstico da geometria e função do VD em uma ampla variedade de doenças 4,5,6,7. Numerosos estudos demonstraram o valor incremental da medida do VD, mesmo usando parâmetros convencionais relativamente simples, o que destaca a importância e a necessidade de quantificação mais precisa da câmara com valor clínico potencialmente significativo.
A ecocardiografia 3D supera várias limitações da avaliação 2D das câmaras cardíacas. Embora a mensuração de volumes e também parâmetros funcionais livres de pressupostos geométricos possa ser de grande interesse também no caso do VE, ela pode ganhar particular importância na avaliação do VD8. Os volumes de VD derivados de 3D e a fração de ejeção (FE) apresentam valor prognóstico significativo em diversas condições cardiovasculares 9,10.
Atualmente, diversos fornecedores fornecem soluções semi-automatizadas para avaliação de RV 3D com resultados validados em relação às medidas de ressonância magnética cardíaca (RM) padrão-ouro11,12. Os requisitos técnicos da avaliação 3D são partes essenciais de um departamento de imagem cardiovascular de última geração hoje em dia, e espera-se que em breve faça parte do equipamento geral em todos os laboratórios de ecocardiografia. Com experiência adequada em aquisição 3D e pós-processamento, a análise 3D RV pode ser facilmente implementada no protocolo de exame padrão.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>3V-D/4V-D/4Vc-D</td>
			<td>General Electric</td>
			<td>n.a.</td>
			<td>ultrasound probe</td>
		</tr>
		<tr>
			<td>4D Auto RVQ</td>
			<td>General Electric</td>
			<td>n.a.</td>
			<td>software for analysis</td>
		</tr>
		<tr>
			<td>E9/E95</td>
			<td>General Electric</td>
			<td>n.a.</td>
			<td>ultrasound machine</td>
		</tr>
		<tr>
			<td>EchoPac v203</td>
			<td>General Electric</td>
			<td>n.a.</td>
			<td>software for analysis</td>
		</tr>
	</tbody>
</table>

morphological and functional assessment of the right ventricle using 3d echocardiography

Tradicionalmente, acreditava-se que o lado direito do coração tem um papel menor na circulação; no entanto, mais e mais dados sugerem que a função do ventrículo direito (VD) tem forte poder diagnóstico e prognóstico em vários distúrbios cardiovasculares. Devido à sua complexa morfologia e função, a avaliação do VD por ecocardiografia bidimensional convencional é limitada: a prática clínica cotidiana geralmente depende de dimensões lineares simples e medidas funcionais. A ecocardiografia tridimensional (3D) superou essas limitações, fornecendo quantificação volumétrica do VD livre de pressupostos geométricos. Aqui, oferecemos um guia passo-a-passo para obter e analisar dados ecocardiográficos 3D do VD usando o principal software comercialmente disponível. Vamos quantificar os volumes de RV 3D e a fração de ejeção. Vários aspectos técnicos podem ajudar a melhorar a qualidade da aquisição e análise de RV também, que apresentamos de forma prática. Revisamos as oportunidades atuais e os fatores limitantes desse método e também destacamos as potenciais aplicações da avaliação de RV 3D na prática clínica atual.

A análise 3D do VD é viável em uma ampla variedade de doenças cardiovasculares. O caso 1 é um voluntário saudável, com volumes e função ventricular normais (Figura 1). O caso 2 é um paciente pós-reparo da valva mitral que é um exemplo típico para os resultados conflitantes da avaliação 2D convencional: enquanto o TAPSE é acentuadamente reduzido, o paciente não apresenta sinais de disfunção do VD e uma função sistólica global do VD mantid...

Watch this Scientific Journal Video about Morphological and Functional Assessment of the Right Ventricle Using 3D Echocardiography at JoVE.com

Morphological and Functional Assessment of the Right Ventricle Using 3D Echocardiography

Aqui, fornecemos um protocolo passo-a-passo de aquisição e análise para a avaliação volumétrica 3D do ventrículo direito, focando principalmente nos aspectos práticos que maximizam a viabilidade dessa técnica.

Avaliação Morfológica e Funcional do Ventrículo Direito Utilizando Ecocardiografia 3D

Traditionally, it was believed that the right side of the heart has a minor role in circulation; however, more and more data suggest that right ...

morphological-functional-assessment-right-ventricle-using-3d

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JoVE Journal

Medicine

2.7K visualizações.  Semmelweis University Heart and Vascular Center. Aqui, fornecemos um protocolo passo-a-passo de aquisição e análise para a avaliação volumétrica 3D do ventrículo direito, focando principalmente nos aspectos práticos que maximizam a viabilidade dessa técnica.

Vídeo: Morphological and Functional Assessment of the Right Ventricle Using 3D Echocardiography

Transthoracic Echocardiography in Mice

In recent years, murine models have become the primary avenue for studying the molecular mechanisms of cardiac dysfunction resulting from changes in gene expression. Transgenic and gene targeting methods can be used to generate mice with altered cardiac size and function,1-3 and as a result, in vivo techniques are needed to evaluate their cardiac phenotype. Transthoracic echocardiography, pulse wave Doppler (PWD), and tissue Doppler imaging (TDI) can be used to provide dimensional measurements of the mouse heart and to quantify the degree of cardiac systolic and diastolic performance. Two-dimensional imaging is used to detect abnormal anatomy or movements of the left ventricle, whereas M-mode echo is used for quantification of cardiac dimensions and contractility.4,5 In addition, PWD is used to quantify localized velocity of turbulent flow,6 whereas TDI is used to measure the velocity of myocardial motion.7 Thus, transthoracic echocardiography offers a comprehensive method for the noninvasive evaluation of cardiac function in mice.

Transthoracic echocardiography offers a noninvasive method for the evaluation of cardiac function in mice. A combination of ultrasound and Doppler imaging modalities can be used to obtain dimensional measurements of the heart and intracardiac blood flow, which together provide an assessment of cardiac systolic and diastolic performance.

Ecocardiografia transtorácica em Ratos

In recent years, murine models have become the primary avenue for studying the molecular mechanisms of cardiac dysfunction resulting from changes in gene ...

Medicina

Echocardiographic Assessment of the Right Heart in Mice

Transgenic and toxic models of pulmonary arterial hypertension (PAH) are widely used to study the pathophysiology of PAH and to investigate potential therapies. Given the expense and time involved in creating animal models of disease, it is critical that researchers have tools to accurately assess phenotypic expression of disease. Right ventricular dysfunction is the major manifestation of pulmonary hypertension. Echocardiography is the mainstay of the noninvasive assessment of right ventricular function in rodent models and has the advantage of clear translation to humans in whom the same tool is used. Published echocardiography protocols in murine models of PAH are lacking.
In this article, we describe a protocol for assessing RV and pulmonary vascular function in a mouse model of PAH with a dominant negative BMPRII mutation; however, this protocol is applicable to any diseases affecting the pulmonary vasculature or right heart. We provide a detailed description of animal preparation, image acquisition and hemodynamic calculation of stroke volume, cardiac output and an estimate of pulmonary artery pressure.

This article provides a protocol for the echocardiographic assessment of right ventricular size and pulmonary hypertension in mice. Applications include phenotype determination and serial assessment in transgenic and toxin-induced mouse models of cardiomyopathy and pulmonary vascular disease.

Avaliação ecocardiográfica do coração direito em Ratos

Transgenic and toxic models of pulmonary arterial hypertension (PAH) are widely used to study the pathophysiology of PAH and to investigate potential ...

Assessment of Right Ventricular Structure and Function in Mouse Model of Pulmonary Artery Constriction by Transthoracic Echocardiography

Emerging clinical data support the notion that RV dysfunction is critical to the pathogenesis of cardiovascular disease and heart failure1-3. Moreover, the RV is significantly affected in pulmonary diseases such as pulmonary artery hypertension (PAH). In addition, the RV is remarkably sensitive to cardiac pathologies, including left ventricular (LV) dysfunction, valvular disease or RV infarction4. To understand the role of RV in the pathogenesis of cardiac diseases, a reliable and noninvasive method to access the RV structurally and functionally is essential.
A noninvasive trans-thoracic echocardiography (TTE) based methodology was established and validated for monitoring dynamic changes in RV structure and function in adult mice. To impose RV stress, we employed a surgical model of pulmonary artery constriction (PAC) and measured the RV response over a 7-day period using a high-frequency ultrasound microimaging system. Sham operated mice were used as controls. Images were acquired in lightly anesthetized mice at baseline (before surgery), day 0 (immediately post-surgery), day 3, and day 7 (post-surgery). Data was analyzed offline using software.
Several acoustic windows (B, M, and Color Doppler modes), which can be consistently obtained in mice, allowed for reliable and reproducible measurement of RV structure (including RV wall thickness, end-diastolic&#160;and end-systolic dimensions), and function (fractional area change, fractional shortening, PA peak velocity, and peak pressure gradient) in normal mice and following PAC.
Using this method, the pressure-gradient resulting from PAC was accurately measured in real-time using Color Doppler mode and was comparable to direct pressure measurements performed with a Millar high-fidelity microtip catheter. Taken together, these data demonstrate that RV measurements obtained from various complimentary views using echocardiography are reliable, reproducible and can provide insights regarding RV structure and function. This method will enable a better understanding of the role of RV cardiac dysfunction.

Right ventricle (RV) dysfunction is critical to the pathogenesis of cardiovascular disease, yet limited methodologies are available for its evaluation. Recent advances in ultrasound imaging provide a noninvasive and accurate option for longitudinal RV study. Herein, we detail a step-by-step echocardiographic method using a murine model of RV pressure overload.

Avaliação da Estrutura e Função do Ventrículo Direito em modelo de camundongo Pulmonary Artery constrição por Ecocardiografia transtorácica

Emerging clinical data support the notion that RV dysfunction is critical to the pathogenesis of cardiovascular disease and heart failure1-3. Moreover, ...

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 do Miocárdio e Avaliação de Resultados Funcional em porcos

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

Chronic Thromboembolic Pulmonary Hypertension and Assessment of Right Ventricular Function in the Piglet

An original piglet model of Chronic Thromboembolic Pulmonary Hypertension (CTEPH) associated with chronic Right Ventricular (RV) dysfunction is described. Pulmonary Hypertension (PH) was induced in 3-week-old piglets by a progressive obstruction of the pulmonary vascular bed. A ligation of the left Pulmonary Artery (PA) was performed first through a mini-thoracotomy. Second, weekly embolizations of the right lower pulmonary lobe were done under fluoroscopic guidance with n-butyl-2-cyanoacrylate during 5 weeks. Mean Pulmonary Arterial Pressure (mPAP) measured by ritght heart catheterism, increased progressively, as well as Right Atrial pressure and Pulmonary Vascular Resistances (PVR) after 5 weeks compared to sham animals. Right Ventricular (RV) structural and functional remodeling were assessed by transthoracic echocardiography (RV diameters, RV wall thickness, RV systolic function). RV elastance and RV-pulmonary coupling were assessed by Pressure-Volume Loops (PVL) analysis with conductance method. Histologic study of the lung and the right ventricle were also performed. Molecular analyses on RV fresh tissues could be performed through repeated transcutaneous endomyocardial biopsies. Pulmonary microvascular disease in obstructed and unobstructed territories was studied from lung biopsies using molecular analyses and pathology. Furthermore, the reliability and the reproducibility was associated with a range of PH severity in animals. Most aspects of the human CTEPH disease were reproduced in this model, which allows new perspectives for the understanding of the underlying mechanisms (mitochondria, inflammation) and new therapeutic approaches (targeted, cellular or gene therapies) of the overloaded right ventricle but also pulmonary microvascular disease.

Chronic Thromboembolic Pulmonary Hypertension (CTEPH) and Right Ventricular (RV) dysfunction were induced in piglets by progressive obstruction of the pulmonary arteries. Consequences were remarkably similar to those observed in CTEPH patients. This animal model would be a very useful tool for pathophysiology and therapeutic experiments on CTEPH and RV failure.

Tromboembólica crônica Hipertensão Pulmonar e Avaliação da Função Ventricular Direita no Piglet

An original piglet model of Chronic Thromboembolic Pulmonary Hypertension (CTEPH) associated with chronic Right Ventricular (RV) dysfunction is described. ...

Assessment of Cardiac Morphological and Functional Changes in Mouse Model of Transverse Aortic Constriction by Echocardiographic Imaging

Transverse aortic constriction (TAC) in mice has been used as a valuable model to study mechanisms of cardiac hypertrophy and heart failure1. A reliable noninvasive method is essential to assess real-time cardiac morphological and functional changes in animal models of heart disease. Transthoracic echocardiography represents an important tool for noninvasive assessment of cardiac structure and function2. Here we used a high-resolution ultrasound imaging system to monitor myocardial remodeling and heart failure progression over time in a mouse model of TAC. B-mode, M-mode, and Doppler imaging were used to precisely assess cardiac hypertrophy, ventricular dilatation, and functional deterioration in mice following TAC. Color and pulse wave (PW) Doppler imaging was used to noninvasively measure pressure gradient across the aortic constriction created by TAC and to assess transmitral blood flow in mice. Thus transthoracic echocardiographic imaging provides comprehensive noninvasive measurements of cardiac dimensions and function in mouse models of heart disease.

The goal of this protocol is to noninvasively assess cardiac structural and functional changes in a mouse model of heart disease created by transverse aortic constriction, using B- and M-mode echocardiography and color/pulse wave Doppler imaging.

Avaliação da morfológicas e funcionais cardíacas Mudanças no Modelo rato de Transverse aórtica constrição por ecocardiográfica de imagem

Transverse aortic constriction (TAC) in mice has been used as a valuable model to study mechanisms of cardiac hypertrophy and heart failure1. A reliable ...

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.

Transtorácica salpico de rastreamento A ecocardiografia para a avaliação quantitativa do Ventrículo Esquerdo do miocárdio Deformação

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

Assessment of the Anticoagulant and Anti-inflammatory Properties of Endothelial Cells Using 3D Cell Culture and Non-anticoagulated Whole Blood

In vivo, endothelial cells are crucial for the natural anticoagulation of circulating blood. Consequently, endothelial cell activation leads to blood coagulation. This phenomenon is observed in many clinical situations, like organ transplantation in the presence of pre-formed anti-donor antibodies, including xenotransplantation, as well as in ischemia/reperfusion injury. In order to reduce animal experimentation according to the 3R standards (reduction, replacement and refinement), in vitro models to study the effect of endothelial cell activation on blood coagulation would be highly desirable. However, common flatbed systems of endothelial cell culture provide a surface-to-volume ratio of 1 - 5 cm2 of endothelium per mL of blood, which is not sufficient for natural, endothelial-mediated anticoagulation. Culturing endothelial cells on microcarrier beads may increase the surface-to-volume ratio to 40 - 160 cm2/mL. This increased ratio is sufficient to ensure the &#34;natural&#34; anticoagulation of whole blood, so that the use of anticoagulants can be avoided. Here an in vitro microcarrier-based system is described to study the effects of genetic modification of porcine endothelial cells on coagulation of whole, non-anticoagulated human blood. In the described assay, primary porcine aortic endothelial cells, either wild type (WT) or transgenic for human CD46 and thrombomodulin, were grown on microcarrier beads and then exposed to freshly drawn non-anticoagulated human blood. This model allows for the measurement and quantification of cytokine release as well as activation markers of complement and coagulation in the blood plasma. In addition, imaging of activated endothelial cell and deposition of immunoglobulins, complement- and coagulation proteins on the endothelialized beads were performed by confocal microscopy. This assay can also be used to test drugs which are supposed to prevent endothelial cell activation and, thus, coagulation. On top of its potential to reduce the number of animals used for such investigations, the described assay is easy to perform and consistently reproducible.

We present an in vitro model which allows the study and analysis of coagulation in whole, non-anticoagulated blood. Anticoagulation in the system depends on the natural anticoagulation effect of healthy endothelial cells and endothelial cell activation will result in clotting.

Avaliação das propriedades das células endoteliais utilizando cultura de células 3D e sangue não anticoagulado anticoagulantes e anti-inflamatório

In vivo, endothelial cells are crucial for the natural anticoagulation of circulating blood. Consequently, endothelial cell activation leads to blood ...

2D and 3D Echocardiography in the Axolotl (Ambystoma Mexicanum)

Cardiac malfunction as a result of ischemic heart disease is a major challenge, and regenerative therapies to the heart are in high demand. A few model species such as zebrafish and salamanders that are capable of intrinsic heart regeneration hold promise for future regenerative therapies for human patients. To evaluate the outcome of cardioregenerative experiments it is imperative that heart function can be monitored. The axolotl salamander (A. mexicanum) represents a well-established model species in regenerative biology attaining sizes that allows for evaluation of cardiac function. The purpose of this protocol is to establish methods to reproducibly measure cardiac function in the axolotl using echocardiography. The application of different anesthetics (benzocaine, MS-222, and propofol) is demonstrated, and the acquisition of two-dimensional (2D) echocardiographic data in both anesthetized and unanesthetized axolotls is described. 2D echocardiography of the three-dimensional (3D) heart can suffer from imprecision and subjectivity of measurements, and to alleviate this phenomenon a solid method, namely intra/inter-operator/observer analysis, to measure and minimize this bias is demonstrated. Finally, a method to acquire 3D echocardiographic data of the beating axolotl heart at a very high spatiotemporal resolution and with pronounced blood-to-tissue contrast is described. Overall, this protocol should provide the necessary methods to evaluate cardiac function and model anatomy, and flow dynamics in the axolotl using ultrasound imaging with applications in both regenerative biology and general physiological experiments.

2D and 3D Echocardiography in the Axolotl Ambystoma Mexicanum

Here we present echocardiography protocols for two-dimensional and three-dimensional image acquisition of the beating heart of the axolotl salamander (Ambystoma mexicanum), a model species in heart regeneration. These methods allow for longitudinal evaluation of cardiac function at a high spatiotemporal resolution.

Ecocardiografia 2D e 3D no Axolotl (Ambystoma Mexicanum)

Cardiac malfunction as a result of ischemic heart disease is a major challenge, and regenerative therapies to the heart are in high demand. A few model ...

Invasive Hemodynamic Assessment for the Right Ventricular System and Hypoxia-Induced Pulmonary Arterial Hypertension in Mice

Pulmonary arterial hypertension (PAH) is a chronic and severe cardiopulmonary disorder. Mice are a popular animal model used to mimic this disease. However, the evaluation of right ventricular pressure (RVP) and pulmonary artery pressure (PAP) remains technically challenging in mice. RVP and PAP are more difficult to measure than left ventricular pressure because of the anatomical differences between the left and right heart systems. In this paper, we describe a stable right heart hemodynamic measurement method and its validation using healthy and PAH mice. This method is based on open-chest surgery and mechanical ventilation support. It is a complicated procedure compared to closed chest procedures. While a well-trained surgeon is required for this surgery, the advantage of this procedure is that it can generate both RVP and PAP parameters at the same time, so it is a preferable procedure for the evaluation of PAH models.

Here, we present a protocol to perform an invasive hemodynamic assessment of the right ventricle and pulmonary artery in mice using an open-chest surgery approach.

Avaliação Hemodinâmica Invasiva para o Sistema Ventricular Direito e hipertensão arterial pulmonar induzida por hipóxia em camundongos

Pulmonary arterial hypertension (PAH) is a chronic and severe cardiopulmonary disorder. Mice are a popular animal model used to mimic this disease. ...