저자는 동물 작업에 도움을 준 국립 실험실 동물 센터의 Minna Törrönen, Riikka Venäläinen, Heikki Karhunen 및 Inkeri Niemi에게 감사를 표합니다. 이 연구는 Finnish Academy, ERC 및 CardioReGenix EU Horizon 보조금의 지원을 받습니다.

모든 실험은 핀란드 동부 대학의 동물 실험위원회의 승인을 받았습니다. 이 프로토콜은 심장 질환에 대한 새로운 치료법의 안전성과 효능을 평가하기 위해 돼지를 위한 임상적으로 관련된 트레드밀 운동 테스트를 설명합니다. 체중 25-80kg의 암컷 국내 돼지를 본 연구에 사용하였다. 동물들은 상업적 공급원으로부터 입수하였다 ( 재료의 표 참조).
1. 러닝 트랙 설정
<ol><li>동물이 한 방향으로만 움직일 수 있도록 달리기 트랙을 설정합니다. 동물이 뒤로 물러나지 않도록 게이트와 해치를 사용하십시오. 달리기 트랙의 평면도는 그림 1에 나와 있으며 달리기 트랙의 예는 그림 2에 나와 있습니다.</li>
	<li>트레드밀( 재료 표 참조)에 경사 변경을 허용할 수 있는 충분한 공간이 있는지 확인하십시오.</li>
	<li>달리는 동안 동물이 회전하는 것을 방지하기 위해 트레드밀의 너비를 조절할 수 있는지 확인하십시오.</li>
	<li>투명 플라스틱을 사용하여 런닝 머신의 전면 벽을 만드십시오. 이것은 동물이 러닝 머신에서 도망가는 것을 방지하지만 여전히 동물이 벽을 통해 볼 수 있도록합니다. 알림: 우리의 경험에 따르면 돼지가 벽 반대편에 있는 동료 돼지를 보면 달리고 싶은 동기가 더 강하기 때문에 동물이 앞 벽을 통해 볼 수 있어야 합니다.</li>
	<li>ECG 모니터와 제세동기( 재료 표 참조)를 런닝머신 옆에 놓습니다. 참고 : 치명적인 부정맥은 특히 돼지가 심근 허혈14,15,16을 앓고있는 경우 스트레스 테스트 중에 발생할 수 있습니다.</li>
	<li>달리기 트랙에 동물들이 달리기 후에 물을 마시고 더위를 식힐 수 있는 물 지점이 있는지 확인하십시오.</li>
</ol>2. 시험 전 돼지의 순응 기간
<ol><li>실험을 시작하기 전에 2주 동안 동물을 수용합니다.</li>
	<li>적응 1 주 동안 동물들이 달리기 트랙을 제외한 조련사와 새로운 주거 환경에 익숙해 지도록하십시오.</li>
	<li>적응 기간의 2 주 동안 동물들이 달리기 트랙에 익숙해 지도록하십시오.</li>
	<li>동물들이 달리기 트랙에 익숙해 지도록 익숙해지십시오. 첫째, 동물들이 트랙을 자유롭게 걷고 환경을 탐험 할 수 있도록 모든 문을 열어 두십시오.</li>
	<li>동물들이 트랙에 더 익숙해지면 러닝머신을 켜고 동물이 한 번에 짧은 시간(예: 7분) 동안 달리게 하십시오. 실행 시간의 길이는 매일 연장되어야 합니다. 참고: 적응 기간 동안 동물에게 보상하는 것을 잊지 마십시오. 예를 들어, 본 연구에서 돼지는 무염 팝콘으로 보상을 받았습니다.</li>
</ol>3. 운동 테스트
알림: 돼지는 운동 테스트 최소 2시간 전에 금식하거나 달리기 전에 소량의 음식만 제공해야 합니다.
<ol><li>트레드밀을 켜고 경사를 5%-10%로 설정합니다.</li>
	<li>동물이 러닝머신에 오르자마자 시작 속도 2km/h로 런닝머신을 시작합니다.</li>
	<li>0.5km/h에 도달할 때까지 60초마다 5km/h씩 속도를 높입니다. 총 실행 시간은 15분입니다.</li>
	<li>동물이 최대 속도로 전체 시간을 달릴 수없는 경우 아래 단계를 수행하십시오.<ol><li>돼지가 선택한 속도만큼 빠르게 달리지 않으면 동물에게 속도를 늦추지 않고 더 빨리 달려야 한다는 느낌을 줄 수 있으므로 뒤에서 부드럽게 밉니다.</li>
		<li>동물을 최대 세 번 부드럽게 밀어보십시오. 그런 다음 돼지가 속도를 감당할 수 있을 때까지 한 번에 0.5km/h씩 속도를 늦춥니다. 2km/h 이하로 감속하지 마십시오.</li>
		<li>동물이 느린 속도로도 달리기를 거부하면 러닝 머신을 끄고 테스트를 중단하십시오.</li>
	</ol></li>
</ol>4. 운동 테스트 중 ECG 모니터링
<ol><li>ECG 전극( 재료 표 참조)을 견갑이나 가슴과 같이 달리는 동안 움직임이 최소화되는 해부학적 위치에 배치합니다. 알림: 운동 테스트용으로 설계된 ECG 전극을 사용하여 피부에 더 잘 접착되도록 하십시오. ECG 전극이 배치될 부위의 머리카락을 면도하는 것을 잊지 마십시오.</li>
	<li>달리는 동안 심박수 변화를 기록합니다. 참고: 우리의 경험에 따르면 ST 세그먼트 분석은 종종 움직임 및 기타 아티팩트로 인해 복잡합니다. 리듬 모니터링은 이식형 루프 레코더 또는 심박 조율기를 사용하여 수행할 수도 있습니다.</li>
</ol>5. 데이터 수집
<ol><li>속도가 바뀔 때마다 주행 거리, 총 시간 및 속도를 기록합니다. 알림: 최신 트레드밀은 다른 많은 데이터를 수집할 수 있으므로 장비의 잠재력을 최대한 활용하려면 트레드밀 설명서를 숙지하는 것이 중요합니다.</li>
	<li>절뚝거림과 같은 동물 행동의 가능한 변화에 주목하십시오. 알림: 필요한 경우 수의사에게 연락하여 동물이 필요한 진통제를 받는지 확인하십시오. 완전히 회복 될 때까지 향후 운동에서 동물을 제거하십시오.</li>
</ol>6. 시술 후 관리
<ol><li>동물이 물 지점에 접근 할 수 있는지 확인하십시오.</li>
	<li>예를 들어 간식이나 장난감으로 동물에게 보상하십시오.</li>
	<li>가능한 부작용에 대해 실행 후 30 분 동안 동물을 모니터링하십시오.</li>
</ol>

운동을 한 돼지의 운동 내성 평가

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저자는 이해 상충을 선언하지 않습니다.

이 대규모 동물 운동 테스트는 클리닉에서 사용되는 테스트를 모방하여 전임상 연구와 임상 시험 간의 종점 격차를 줄입니다. 폐쇄성 동맥경화증, 심부전, 허혈성 심장질환 등 중증 심혈관질환에 대한 새로운 치료법의 효능을 평가하는 데 적용할 수 있다. 이 프로토콜에 적용되는 시점은 테스트된 치료에 따라 다를 수 있습니다. 이 프로토콜은 대형 동물에 대한 오랜 경험을 바탕으로 표준화되었?...

만성 심혈관 질환은 전 세계적으로 사망률과 이환율의 중요한 원인입니다 1,2. 현재의 치료법이 대다수의 환자에게 효과적이지만, 예를 들어 확산성 만성 질환이나 동반 질환으로 인해 많은 사람들이 여전히 현재 치료법의 혜택을 받을 수 없습니다. 또한, 일부 환자의 경우 이용 가능한 치료법으로 심장 증상이 완화되지 않고 최적의 의학적 치료에도 불구하고 심혈관 질환이 진행된다3. 따라서 중증 심혈관 질환에 대한 새로운 치료 옵션을 개발할 필요성이 분명합니다.
지난 몇 년 동안 이러한 표적을 조작하는 새로운 분자 경로와 방법이 발견되어 유전자 치료, 세포 치료 및 기타 새로운 치료법이 중증 심혈관 질환 치료를 위한 현실적인 옵션이 되었습니다4. 그러나 유망한 전임상 결과 이후 많은 심혈관 응용 프로그램이 임상 시험에서 기대치를 충족시키지 못했습니다. 임상 시험에서 낮은 효능에도 불구하고, 몇몇 시험은 새로운 치료법 5,6,7,8,9의 우수한 안전성 프로파일을 확립했다. 따라서 환자에게 새로운 심혈관 요법을 제공하려면 임상적 효능을 예측할 수 있는 전임상 연구에서 개선된 접근 방식과 더 나은 전임상 모델, 연구 환경 및 종점이 필요합니다.
동물 모델에서 강조점은 일반적으로 조직학적 절편에서 계산된 모세혈관의 수와 면적 또는 휴식 시 및 약리학적 스트레스 하에서 좌심실 영상의 매개변수와 같이 쉽게 정량화할 수 있는 종점에 있었습니다. 임상 시험에서 운동 내성 또는 증상 완화와 같은 많은 평가변수가 보다 주관적이었다4. 따라서 전임상 연구 및 임상 시험의 종점은 응용 요법의 다양한 측면을 측정할 가능성이 높습니다. 예를 들어, 혈관 양의 증가가 항상 더 나은 관류, 심장 기능 또는 운동 내성과 관련이 있는 것은 아닙니다. 그럼에도 불구하고, 성공적인 치료적 접근법을 개발하기 위해서는 두 가지 유형의 종점모두 필요하다 10. 그러나 주요 목표는 항상 증상을 완화하고 환자의 예후와 삶의 질을 향상시키는 것입니다. 이를 위해서는 전임상 연구와 임상 연구 간에 종말점 측정이 더 잘 일치해야 한다4.
심폐 건강은 지속적인 신체 활동 중에 산소를 공급하는 순환계 및 호흡계의 능력을 반영하므로 개인의 기능적 능력을 정량화합니다. 기능적 능력은 심혈관 및 모든 원인으로 인한 사망 위험에 대한 강력하고 독립적인 예측 인자이기 때문에 주요 예후 지표이다11. 심폐 기능의 개선은 사망 위험 감소와 관련이 있다12. 운동 검사는 심혈관 질환의 유산소 성능 및 치료 반응을 평가하는 데 적합합니다. 가용성에 따라 테스트는 자전거 에르고 미터 또는 러닝 머신에서 수행됩니다. 분당 작업량의 점진적인 증가가 일반적으로 사용되며 급격한 증가는 피할 수 있습니다. 이것은 선형 생리적 반응으로 이어집니다. 운동 테스트에서 가장 중요한 변수에는 총 운동 시간, 달성한 대사 등가물(MET), 심박수, QRS 복합체(Q, R 및 S파)와 T파(ST 세그먼트) 사이의 심전도(ECG) 라인의 변화가 포함됩니다. 임상적 스트레스 테스트는 비용이 저렴하고 쉽게 접근할 수 있다13. 이러한 이유로 6분 걷기 테스트와 같은 스트레스 테스트는 클리닉에서 널리 사용되어 왔으며 새로운 치료법의 전임상 평가에도 사용해야 합니다.
우리가 아는 한, 유전자 요법 또는 다른 새로운 치료법의 기능적 효능을 평가하기 위한 잘 설명된 대형 동물 모델은 없습니다. 따라서 임상적으로 관련된 운동 테스트는 전임상 환경에서 이러한 새로운 치료법의 효율성을 평가하기 위한 훌륭한 관점을 제공합니다.

<table><tbody><tr><td>Defibrillator</td><td>Zoll M series</td><td>TO9K116790</td><td>All portable defribrillators will work</td></tr><tr><td>Defibrillator pads</td><td>Philips</td><td>M3713A</td><td>All pads work, as long as the pads are compatible with the defibrillator</td></tr><tr><td>ECG electrodes</td><td>Several providers</td><td></td><td>Prefer ECG electrodes designed for exercise tests</td></tr><tr><td>Loop recorder</td><td>Abbott Oy</td><td>DM3500</td><td>Optional for rhythm monitoring</td></tr><tr><td>Patient monitor</td><td>Schiller Argus LCM Plus</td><td>7,80,05,935</td><td>All portable ecg monitors will work</td></tr><tr><td>Pigs</td><td>Emolandia Oy</td><td></td><td></td></tr><tr><td>Treadmill</td><td>NordicTrack</td><td></td><td>All treadmills with adjustable incline and speed are suitable for the exercise test.&amp;nbsp; The treadmill should be as long and wide as possible.</td></tr><tr><td>Ultrasound system</td><td>Philips EPIQ 7 ultrasound</td><td></td><td></td></tr><tr><td>Various building materials</td><td>Several providers</td><td></td><td>For building fences, ramps and gates according to the Figure 1 and Figure 2</td></tr><tr><td>Various treats for the animals</td><td></td><td></td><td></td></tr></tbody></table>

exercise test for evaluation of the functional efficacy of the pig cardiovascular system

치료의 발전에도 불구하고 심혈관 질환은 여전히 전 세계적으로 사망률과 이환율의 가장 큰 원인 중 하나입니다. 유전자 치료 기반 치료 혈관신생은 최적의 약물 요법과 침습적 절차에도 불구하고 심각한 증상이 있는 환자를 치료하기 위한 유망한 접근 방식입니다. 그러나 많은 유망한 심혈관 유전자 치료 기술은 임상 시험에서 기대치를 달성하지 못했습니다. 한 가지 설명은 효능을 측정하는 데 사용되는 전임상 및 임상 종점 간의 불일치입니다. 동물 모델에서는 일반적으로 조직학적 절편에서 계산된 모세혈관의 수 및 면적과 같이 쉽게 정량화할 수 있는 종말점에 중점을 두었습니다. 사망률과 이환율 외에도 임상 시험의 종점은 운동 내성 및 삶의 질과 같은 주관적입니다. 그러나 전임상 및 임상 종점은 응용 요법의 다른 측면을 측정할 가능성이 높습니다. 그럼에도 불구하고 성공적인 치료 접근법을 개발하려면 두 가지 유형의 종점이 모두 필요합니다. 클리닉에서 주요 목표는 항상 환자의 증상을 완화하고 예후와 삶의 질을 향상시키는 것입니다. 전임상 연구에서 더 나은 예측 데이터를 얻으려면 종점 측정이 임상 연구의 측정과 더 잘 일치해야 합니다. 여기에서는 돼지의 임상적으로 관련된 트레드밀 운동 테스트를 위한 프로토콜을 소개합니다. 이 연구의 목표는 (1) 유전자 요법 및 기타 새로운 치료법의 안전성과 기능적 효능을 평가하는 데 사용할 수 있는 돼지에 대한 신뢰할 수 있는 운동 테스트를 제공하고 (2) 전임상 연구와 임상 연구 간의 종점을 더 잘 일치시키는 것입니다.

이 프로토콜로 성공하려면 큰 동물과 함께 일한 경험이 있어야 합니다. 연구자들은 동물이 피로 또는 동기 부여 부족으로 인해 달리기를 멈추는지 평가할 수 있어야 합니다. 속도와 거리를 기록하면 이를 평가하는 데 도움이 될 수 있는데, 일반적으로 동기 부여가 부족한 동물은 달리기를 완전히 멈추는 반면 피곤한 동물은 속도를 늦춘 후에도 계속 달리기를 하기 때문입니다(그림...

Watch this Scientific Journal Video about Exercise Test for Evaluation of the Functional Efficacy of the Pig Cardiovascular System at JoVE.com

저자 스포트라이트: Pig Cardiovascular System의 기능적 효능 평가를 위한 운동 테스트  

본 프로토콜은 전임상 환경에서 새로운 치료법의 효율성을 평가하기 위한 심혈관계의 기능적 능력을 평가하기 위한 대규모 동물 운동 테스트 모델을 설명합니다. 임상 운동 테스트와 비슷합니다.

돼지 심혈관계 기능적 효능 평가를 위한 운동시험

Despite the progress in treatments, cardiovascular diseases are still one of the biggest causes of mortality and morbidity worldwide. Gene therapy-based ...

이 연구의 목적은 유전자 요법 및 기타 새로운 치료법의 안전성과 기능적 효능을 평가하는 데 사용할 수 있는 신뢰할 수 있는 운동 테스트를 돼지에게 제공하는 것입니다. 일반적으로 심혈관 동물 연구는 조직학적 섹션에서 계산된 모세혈관 면적과 같이 쉽게 정량화할 수 있는 종점을 강조합니다. 대조적으로, 임상 연구는 과도한 내성과 같은 보다 주관적인 종점을 사용합니다.이 프로토콜은 전임상 연구와 임상 연구 간의 종점을 더 잘 일치시킵니다. 이전에 발표된 장기 실행 세션의 방법은 만성 운동 적응을 평가하는 것을 목표로 하는 반면, 제시된 방법은 심폐 건강을 평가하는 데 더 적합합니다.

exercise-test-for-evaluation-functional-efficacy-pig-cardiovascular

Universidad de Cartagena UDEC

Research

JoVE Journal

Genetics

Exercise Test for Evaluation of the Functional Efficacy of the Pig Cardiovascular System

1.2K 조회수.  University of Eastern Finland. 이 연구의 목적은 유전자 요법 및 기타 새로운 치료법의 안전성과 기능적 효능을 평가하는 데 사용할 수 있는 신뢰할 수 있는 운동 테스트를 돼지에게 제공하는 것입니다. 일반적으로 심혈관 동물 연구는 조직학적 섹션에서 계산된 모세혈관 면적과 같이 쉽게 정량화할 수 있는 종점을 강조합니다. 대조적으로, 임상 연구는 과도한 내성과 같은 보다 주관적인 종점을 사용합니...

비디오: 저자 스포트라이트: Pig Cardiovascular System의 기능적 효능 평가를 위한 운동 테스트  

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.

돼지의 심근 경색 및 기능적 결과의 평가

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

연구

의학

An Isolated Working Heart System for Large Animal Models

Since its introduction in the late 19th century, the Langendorff isolated heart perfusion apparatus, and the subsequent development of the working heart model, have been invaluable tools for studying cardiovascular function and disease1-15. Although the Langendorff heart preparation can be used for any mammalian heart, most studies involving this apparatus use small animal models (e.g., mouse, rat, and rabbit) due to the increased complexity of systems for larger mammals1,3,11. One major difficulty is ensuring a constant coronary perfusion pressure over a range of different heart sizes &#8211; a key component of any experiment utilizing this device1,11. By replacing the classic hydrostatic afterload column with a centrifugal pump, the Langendorff working heart apparatus described below allows for easy adjustment and tight regulation of perfusion pressures, meaning the same set-up can be used for various species or heart sizes. Furthermore, this configuration can also seamlessly switch between constant pressure or constant flow during reperfusion, depending on the user&#8217;s preferences. The open nature of this setup, despite making temperature regulation more difficult than other designs, allows for easy collection of effluent and ventricular pressure-volume data.

Most studies involving the Langendorff apparatus use small animal models due to the increased complexity of systems for larger mammals. We describe a Langendorff system for large animal models that allows for use across a range of species, including humans, and relatively easy data acquisition.

큰 동물 모델에 대한 격리 된 작업 심장 시스템

Since its introduction in the late 19th century, the Langendorff isolated heart perfusion apparatus, and the subsequent development of the working heart ...

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.

만성 혈전 색 전성 폐 고혈압과 돼지에서 마우스 오른쪽 심실 기능의 평가

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

Surgical Placement of Catheters for Long-term Cardiovascular Exercise Testing in Swine

This protocol describes the surgical procedure to chronically instrument swine and the procedure to exercise swine on a motor-driven treadmill. Early cardiopulmonary dysfunction is difficult to diagnose, particularly in animal models, as cardiopulmonary function is often measured invasively, requiring anesthesia. As many anesthetic agents are cardiodepressive, subtle changes in cardiovascular function may be masked. In contrast, chronic instrumentation allows for measurement of cardiopulmonary function in the awake state, so that measurements can be obtained under quiet resting conditions, without the effects of anesthesia and acute surgical trauma. Furthermore, when animals are properly trained, measurements can also be obtained during graded treadmill exercise.
Flow probes are placed around the aorta or pulmonary artery for measurement of cardiac output and around the left anterior descending coronary artery for measurement of coronary blood flow. Fluid-filled catheters are implanted in the aorta, pulmonary artery, left atrium, left ventricle and right ventricle for pressure measurement and blood sampling. In addition, a 20 G&#160;catheter is positioned in the anterior interventricular vein to allow coronary venous blood sampling.
After a week of recovery, swine are placed on a motor-driven treadmill, the catheters are connected to pressure and flow meters, and swine are subjected to a five-stage progressive exercise protocol, with each stage lasting 3 min. Hemodynamic signals are continuously recorded and blood samples are taken during the last 30 sec of each exercise stage.
The major advantage of studying chronically instrumented animals is that it allows serial assessment of cardiopulmonary function, not only at rest but also during physical stress such as exercise. Moreover, cardiopulmonary function can be assessed repeatedly during disease development and during chronic treatment, thereby increasing statistical power and hence limiting the number of animals required for a study.

Here we present a protocol to assess cardiopulmonary function in awake swine, at rest and during graded treadmill exercise. Chronic instrumentation allows for repeated hemodynamic measurements uninfluenced by cardiodepressive anesthetic agents.

돼지의 장기 심장 혈관 운동 테스트를위한 카테터의 외과 배치

This protocol describes the surgical procedure to chronically instrument swine and the procedure to exercise swine on a motor-driven treadmill. Early ...

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.

급성 심근 경색의 돼지 모델의 주요 결과 평가

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

Tachycardia-Induced Cardiomyopathy As a Chronic Heart Failure Model in Swine

A stable and reliable model of chronic heart failure is required for many experiments to understand hemodynamics or to test effects of new treatment methods. Here, we present such a model by tachycardia-induced cardiomyopathy, which can be produced by rapid cardiac pacing in swine.
A single pacing lead is introduced transvenously into fully anaesthetized healthy swine, to the apex of the right ventricle, and fixated. Its other end is then tunneled dorsally to the paravertebral region. There, it is connected to an in-house modified heart pacemaker unit that is then implanted in a subcutaneous pocket. 
After 4 - 8 weeks of rapid ventricular pacing at rates of 200 - 240 beats/min, physical examination revealed signs of severe heart failure - tachypnea, spontaneous sinus tachycardia, and fatigue. Echocardiography and X-ray showed dilation of all heart chambers, effusions, and severe systolic dysfunction. These findings correspond well to decompensated dilated cardiomyopathy and are also preserved after the cessation of pacing.
This model of tachycardia-induced cardiomyopathy can be used for studying the pathophysiology of progressive chronic heart failure, especially hemodynamic changes caused by new treatment modalities like mechanical circulatory supports. This methodology is easy to perform and the results are robust and reproducible.

Here, we present a protocol to produce tachycardia-induced cardiomyopathy in swine. This model represents a potent way to study the hemodynamics of progressive chronic heart failure and the effects of applied treatment.

돼지에서 만성 심장 마비 모델로 tachycardia 유발 심근

A stable and reliable model of chronic heart failure is required for many experiments to understand hemodynamics or to test effects of new treatment ...

Optocardiography and Electrophysiology Studies of Ex Vivo Langendorff-perfused Hearts

Small animal models are most commonly used in cardiovascular research due to the availability of genetically modified species and lower cost compared to larger animals. Yet, larger mammals are better suited for translational research questions related to normal cardiac physiology, pathophysiology, and preclinical testing of therapeutic agents. To overcome the technical barriers associated with employing a larger animal model in cardiac research, we describe an approach to measure physiological parameters in an isolated, Langendorff-perfused piglet heart. This approach combines two powerful experimental tools to evaluate the state of the heart: electrophysiology (EP) study and simultaneous optical mapping of transmembrane voltage and intracellular calcium using parameter sensitive dyes (RH237, Rhod2-AM). The described methodologies are well suited for translational studies investigating the cardiac conduction system, alterations in action potential morphology, calcium handling, excitation-contraction coupling and the incidence of cardiac alternans or arrhythmias.

The objective of this study was to establish a method for investigating cardiac dynamics using a translational animal model. The described experimental approach incorporates dual-emission optocardiography in conjunction with an electrophysiological study to assess electrical activity in an isolated, intact porcine heart model.

전 비보 랑엔도르프-퍼블릭 하트의 검문 및 전기생리학 연구

Small animal models are most commonly used in cardiovascular research due to the availability of genetically modified species and lower cost compared to ...

Post-Myocardial Infarction Heart Failure in Closed-chest Coronary Occlusion/Reperfusion Model in G&#246;ttingen Minipigs and Landrace Pigs

The development of heart failure is the most powerful predictor of long-term mortality in patients surviving acute myocardial infarction (MI). There is an unmet clinical need for prevention and therapy of post-myocardial infarction heart failure (post-MI HF). Clinically relevant pig models of post-MI HF are prerequisites for final proof-of-concept studies before entering into clinical trials in drug and medical device development.
Here we aimed to characterize a closed-chest porcine model of post-MI HF in adult G&#246;ttingen minipigs with long-term follow-up including serial cardiac magnetic resonance imaging (CMRI) and to compare it with the commonly used Landrace pig model.
MI was induced by intraluminal balloon occlusion of the left anterior descending coronary artery for 120 min in G&#246;ttingen minipigs and for 90 min in Landrace pigs, followed by reperfusion. CMRI was performed to assess cardiac morphology and function at baseline in both breeds and at 3 and 6 months in G&#246;ttingen minipigs and at 2 months in Landrace pigs, respectively.
Scar sizes were comparable in the two breeds, but MI resulted in a significant decrease of left ventricular ejection fraction (LVEF) only in G&#246;ttingen minipigs, while Landrace pigs did not show a reduction of LVEF. Right ventricular (RV) ejection fraction increased in both breeds despite the negligible RV scar sizes. In contrast to the significant increase of left ventricular end-diastolic (LVED) mass in Landrace pigs at 2 months, G&#246;ttingen minipigs showed a slight increase in LVED mass only at 6 months.
In summary, this is the first characterization of post-MI HF in G&#246;ttingen minipigs in comparison to Landrace pigs, showing that the G&#246;ttingen minipig model reflects post-MI HF parameters comparable to the human pathology. We conclude that the G&#246;ttingen minipig model is superior to the Landrace pig model to study the development of post-MI HF.

The overall goal of the current study is to present the techniques of induction of myocardial infarction (MI) and post-myocardial infarction heart failure (post-MI HF) in closed-chest, adult G&#246;ttingen minipigs and the characterization of post-MI HF model in G&#246;ttingen minipigs as compared to Landrace pigs.

괴팅겐 미니피그와 랜드레이스 돼지의 폐쇄 가슴 관상 동맥 폐색/재관류 모델의 심근 경색 심부전

The development of heart failure is the most powerful predictor of long-term mortality in patients surviving acute myocardial infarction (MI). There is an ...

Measuring Cardiac Output in a Swine Model

Swine are frequently used in medical research given their similar cardiac physiology to that of humans. Measuring cardiac parameters such as stroke volume and cardiac output are essential in this type of research. Contrast ventriculography, thermodilution, and pressure-volume loop (PV-loop) catheters can be used to accurately obtain cardiac performance data depending on which resources and expertise are available. For this study,five Yorkshire swine were anesthetized and intubated. Central venous and arterial access was obtained to place the necessary measurement instruments.A temperature probe was placed in the aortic root. A cold saline bolus was delivered to the right atrium and temperature deflection curve was recorded. Integration of the area under the curve allowed for the calculation of the current cardiac output.A pigtail catheter was percutaneously placed in the left ventricle and 30 mL of iodinated contrast was power injected over 2 seconds. Digital subtraction angiography images were uploaded to volumetric analysis software to calculate the stroke volume and cardiac output. A pressure volume-loop catheter was placed into the left ventricle (LV) and provided continuous pressure and volume data of the LV, which allowed the calculation of both stroke volume and cardiac output.All three methods demonstrated good correlation with each other. The PV-loop catheter and thermodilution exhibited the best correlation with a 3% error and a Pearson coefficient of 0.99, with 95% CI=0.97 to 1.1, (p=0.002). The PV-loop catheter against ventriculography also showed good correlation with a 6% error and a Pearson coefficient of 0.95, 95% CI=0.96 to 1.1 (p=0.01). Finally, thermodilution against ventriculography had a 2% error with r=0.95, 95% CI=0.93 to 1.11, (p=0.01). In conclusion, we state that the PV-loop catheter, contrast ventriculography, and thermodilution each offer certain advantages depending on the researcher's requirements. Each method is reliable and accurate for measuring various cardiac parameters in swine such as the stroke volume and cardiac output.

Thermodilution, pressure-volume loop catheters, and contrast ventriculography are reliable and accurate methods for determining cardiac physiology such as stroke volume and cardiac output in a laboratory setting in swine.

Swine are frequently used in medical research given their similar cardiac physiology to that of humans. Measuring cardiac parameters such as stroke volume ...

Echocardiography Recording in Awake Miniature Pigs

Echocardiography uses ultrasonic waves to non-invasively assess cardiac structure and function and is the standard of care for cardiac assessment and monitoring. The miniature pig, or minipig, is increasingly being used as a model of cardiac disease in medical research. Pigs are notoriously difficult to restrain and handle safely, and, therefore, research echocardiography in this species is almost always performed under anesthesia or heavy sedation. Anesthetics and sedatives universally affect cardiovascular function and may cause the depression of cardiac output and blood pressure, increases or decreases in heart rate and systemic vascular resistance, changes in the electrical rhythm, and altered coronary blood flow. Therefore, sedated or anesthetized echocardiography may not accurately depict the progression of cardiac disease in large animal models, thereby limiting the translational value of these important studies. This paper describes a novel device that allows for standing awake echocardiography in minipigs. In addition, training techniques used to teach pigs to tolerate this painless and non-invasive procedure without the need for hemodynamic-altering anesthetics are described. Standing awake echocardiography represents a safe and feasible way to perform the most common cardiac monitoring test in minipigs for cardiovascular research.

A simple cart construct, built to perform research echocardiography in standing awake minipigs, is described, along with building considerations, training techniques, and representative ultrasound images.

Awake Miniature Pigs의 심 초음파 녹음

Echocardiography uses ultrasonic waves to non-invasively assess cardiac structure and function and is the standard of care for cardiac assessment and ...