이 작품은 NIH R01 HL093183, HL088434, HL071763, HL080498, HL083156 및 P20HL100396 (RJH)에 의해, Caerus 네트워크 (RJH)를 통해 Leducq 재단에서 지원됩니다. DL는 독일 연구 재단에 의해 지원되었다. 우리는이 프로젝트에 그녀의 기술 지원 로렌 Leonardson 감사하고 싶습니다.

1. 심근 경색 생성 <ol><li> 동물 생명 공학 산업 (도일 스타운, PA)에서 여성 요크셔 돼지 (20kg)는 Telazol (tiletamine / zolazepam) (8.0 밀리그램 / kg)과 Buprenorphine를 (0.6 MG)를 사용하여 미리 치료하고 있습니다. </li><li> 동물 100 % 산소와 intubated 후 통풍이 있습니다. 전신 마취는 수술을 통해 Propofol (5-8mg/kg/hr)로 유지됩니다. </li><li> 대퇴 사이트는 povidone 요오드 다음 70% 이소 프로필 알코올로 준비가되어 있습니다. percutaneous의 바늘이 동맥에 접근하고 칼집 배치에 대한을 제공합니다. 칼집 삽입 후, 헤파린의 3000-5000 IU는 2백50~3백초의 활성 응고 시간을 유지하기 위해 IV를 관리합니다. </li><li> Amiodarone (75 MG)의 볼러스 선량은 intramuscularly 관리합니다. Amiodarone (75 MG), 아트로 핀 (2 MG) 및 칼륨 아세테이트 (20 mEq)와 1000 ML 호수 가방 정맥 주사가 시작 및 절차 전반에 걸쳐 150 ML / HR의 속도를 사용하여 유지됩니다. </li><li> 7 천 하키 스틱 카테터는 (Cordis, 마이애미, FL) 왼쪽 관상 동맥 고급이며, 관상 동맥 혈관 조영술이 수행됩니다. 0.014 인치 가이드 와이어는 (애보트, 공원, IL) 관상 동맥을 내림차순 왼쪽 앞쪽에 (내사)에 고급입니다. 8 - mm 길이, 4.0 mm 보이저 이상 - 더 - 와이어 풍선이 (애보트, 공원, IL) 고급 위치 근위 내사과에서, 그리고 관상 동맥 풍선은 3 ATM에 부풀입니다. 그 소년의 폐색은 혈관 촬영으로 확인됩니다. </li><li> 지속적인 심전도 모니터링 및 hemodynamic로 폐색 90 분 동안 유지됩니다. 악성 심실 arrhythmias는 종종 시행하는 경우에는 200 J에서 300 J로 직류 충격 즉시 배달을에, 처음 40 분 내에 발생할 수 있습니다. 보통 부정맥의 두 피크 시간 프레임 하나 둘 사이에 10 분, 풍선 인플레이션 이후 25 35분 사이에 다른있다. 프레임이 표시된 시간 내에 심실 세동을 입력 이상의 95 % 발생률이 있기 때문에 열심히 모니터링 및 즉각적인 defibrillation은 성공적인 심근 경색 생성하는 열쇠입니다. 즉각적이고 적절한 defibrillation이 수행되는 경우, 생존율은 매우 높습니다. 혈압 80 mmHg 이하로 떨어지면, 볼륨 부하가 빠른 정맥 주입 호수로 수행됩니다. </li><li> 풍선 deflated하고 모든 장치가 제거됩니다. </li><li> 칼집은 대퇴 동맥에서 제거, 마취가 원천 징수되고 동물은 복구하는 동안 모니터입니다. Amiodarone, Atropin 및 칼륨 아세테이트 주입과 정맥 호수 50 ML / 시간에서 감소하고 하룻밤 주어집니다. Nitroglycerline 및 Furoscemide의 근육내 주사가 관리합니다. 그러면 동물은 복구할 수 있으며,주의 ECG 모니터링은 최소한 3 시간 동안 계속됩니다. </li><li> 아스피린 (200mg) PO는 5 일 동안 주어집니다. </li></ol> 심근 경색 생성 절차 도중 후 일반적인 합병증은 관상 동맥 절개, 펑크 사이트 혈종, 부정맥 및 심장 마비입니다. 기술을 보완하고 철저하게 모니터링은 이러한 합병증을 예방하고 관리해야합니다. 2. 바이러스 사출 <ol><li> 위의 심근 경색 생성 steps1 - 3에 표시된 같은 동물은 준비가되어 있습니다. 헤파린의 동일한 복용량은 바이러스 주입 중에 혈전증을 방지하기 위해 제공해야합니다. </li><li> 5Fr 하키 스틱 카테터는 (Cordis, 마이애미, FL) 왼쪽 관상 동맥 고급입니다. 혈관 촬영 후, 0.014 인치 가이드 와이어의 두 소년에 하나 카테터의 위치를​​ 해결하기 위해 왼쪽의 곡절 악센트 관상 동맥 (누설 동축 케이블)을 한 고급 있습니다. </li><li> 동맥혈 (-20 ML)은 카테터에서 얻을 수 있습니다. 주입하는 동안 국소 빈혈을 방지하기 위해, 절반은 희석 혈액이 솔루션에 사용됩니다. 바이러스 10 ML 혈액과 혼합과 호수가있는 20 ML로 희석합니다. 나머지 10 ML도 20 ML로 희석하여 카테 테르 루멘 내에 잔류 바이러스를 전달하는 데 사용됩니다. </li><li> 정맥 주사 니트로 글리세린 (20 μg / 분)은 귀 정맥을 통해 시작됩니다. </li><li> 카테 테르 팁은 근위 왼쪽 주요 트랙트에 배치됩니다. 카테터를 통해 압력은 카테 테르가 wedging되지 않도록 감시하고 있습니다. </li><li> 바이러스 혼합 혈액 (15 ML)은 왼쪽 관상 동맥에 주입 펌프를 사용하여 십오분 통해 카테터를 통해 주입합니다. 십오분 후, 주사기가 빠질때로 교환하고, 주입은 10 분 동안 계속됩니다. </li><li> 철사와 카테터는 바로 관상 동맥 왼쪽 관상 동맥과 같은 방법으로 해결됩니다. </li><li> 바이러스와 혼합 혈액의 나머지 5 ML은 물을 10 분 다음 5 분 오른쪽 관상 동맥에 주입합니다. </li></ol> 3. 대표 사례 그림 1은 심근 경색 작성 후 일개월에서 왼쪽 ventriculography를 보여줍니다. 왼쪽 심실 벽의 운동이 크게의 배출 분율 (EF)와 장애인입니다31%. 그림 2는 일개월에서 좌심실의 교차 부분을 보여줍니다. 기지에서 앞쪽에 벽의 꼭대기에 큰 흉터를 볼 수 있습니다. 이 모델에서 심근 경색 후 일개월에서 평균 EF 36 %입니다 그리고 심근 경색없이 같은 크기의 동물 이들의 평균은 각각 68% 42 ML, 반면 평균 끝 diastolic 볼륨 83 ML이다. 그림 3의 왼쪽 심실 국경 지대 지역에서 베타 - 여자의 표현 adeno - 관련 바이러스 (CMV rAAV1 락 - Z 2 X 10 12 DNAse resitant 입자)의 주사 후 1 개월 보여줍니다. 강화된 표현은 바이러스 주입 지역에서 발견되었다. <img src="/files/ftp_upload/2778/2778fig1.jpg" alt="그림 1" /> 그림 1. 분출 분율은 심근 경색 생성 후 일개월 왼쪽 ventriculography에서 측정. 좌심실은 92.5 ML로 확대되고 앞쪽에 벽 움직임이 크게 손상됩니다. <img src="/files/ftp_upload/2778/2778fig2.jpg" alt="그림 2" /> 그림 2. triphenyl tetrazolium 염화 물들어 심근 경색 작성 후 일개월에서 좌심실의 교차 부분은 흉터 영역을 윤곽을 그리다 수 있습니다. 기지에서 앞쪽에 벽의 꼭대기에 큰 흉터를 볼 수 있습니다. <img src="/files/ftp_upload/2778/2778fig3.jpg" alt="그림 3" /> 그림 3. adeno - 관련 바이러스 (CMV rAAV1 락 - Z 2 X 10 12 DNAse 강한 입자)에 의해 유도된 베타 여자의 표현. 강화된 표현은 intracoronary 바이러스 주사 후 경색과 정상 조직, 일개월 사이의 국경 지대 지역에서 발견된다. 오른쪽 하단 사진은 정상적인 생리가 동일한 프로토콜을 사용하여 관상 동맥에 주입했던 부정적인 컨트롤을 보여줍니다.

<ol>
	<li>Perez de Prado, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Closed-chest+experimental+porcine+model+of+acute+myocardial+infarction-reperfusion.">Closed-chest experimental porcine model of acute myocardial infarction-reperfusion.</a> J Pharmacol Toxicol Methods. 60, 301-306 (2009).</li><li>Angeli, F. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Left+ventricular+remodeling+after+myocardial+infarction:+characterization+of+a+swine+model+on+beta-blocker+therapy.">Left ventricular remodeling after myocardial infarction: characterization of a swine model on beta-blocker therapy.</a> Comp Med. 59, 272-279 (2009).</li><li>Urdaneta, F., Lobato, E. B., Kirby, D. S. &. a. m. p. ;. a. m. p., Sidi, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Treating+myocardial+stunning+randomly,+with+either+propofol+or+isoflurane+following+transient+coronary+occlusion+and+reperfusion+in+pigs.">Treating myocardial stunning randomly, with either propofol or isoflurane following transient coronary occlusion and reperfusion in pigs.</a> Ann Card Anaesth. 12, 113-121 (2009).</li><li>Jaski, 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=Calcium+upregulation+by+percutaneous+administration+of+gene+therapy+in+cardiac+disease+(CUPID+Trial),+a+first-in-human+phase+1/2+clinical+trial.">Calcium upregulation by percutaneous administration of gene therapy in cardiac disease (CUPID Trial), a first-in-human phase 1/2 clinical trial.</a> J Card Fail. 15, 171-181 (2009).</li></ol>

모든 저자는 원고와 관련하여 관심의 충돌로 포즈 수도 상업적인 연관이 없습니다.

심장 마비의 다양한 원인 중 국소 빈혈은 멀리 가장 자주 병인에하는 것입니다. 이 심각한 질병을 극복하기 위해 다양한 치료 방법이 개발되고있다. 이러한 유망 치료법이 클리닉에 적용할 수있는 전에 요법의주의 평가가 필요합니다. 따라서, 환자의 임상 상태를 나타냅니다 허혈성 심부전 모델의 확립은 심장 마비의 요법의 발전에 중요합니다. 우리 돼지 심근 경색 모델이 translational 요구 사항?...

<!DOCTYPE html PUBLIC "-//W3C//DTD XHTML 1.0 Transitional//EN" "http://www.w3.org/TR/xhtml1/DTD/xhtml1-transitional.dtd">
<html xmlns="http://www.w3.org/1999/xhtml" xml:lang="en" lang="en">	
		
		<div>
		<table border="1">
		<thead>
		<tr>
		<th>Material Name</th>
		<th>Type</th>
		<th>Company</th>
		<th>Catalogue Number</th>
		<th>Comment</th>
		</tr>
		</thead>
		<tbody>
		
<tr>
<td>Telazol</td>
<td>&nbsp;</td>
<td>Fort Dodge Animal Health</td>
<td>0856-9050-49</td>
<td>&nbsp;</td>
<tr>
<td>Propofol</td>
<td>&nbsp;</td>
<td>APP Pharmaceuticals</td>
<td>63323-270-20</td>
<td>&nbsp;</td>
<tr>
<td>Heparin</td>
<td>&nbsp;</td>
<td>APP Pharmaceuticals</td>
<td>63323-540-11</td>
<td>&nbsp;</td>
<tr>
<td>Amiodarone</td>
<td>&nbsp;</td>
<td>APP Pharmaceuticals</td>
<td>63323-616-03</td>
<td>&nbsp;</td>
<tr>
<td>Atropin Sulfate</td>
<td>&nbsp;</td>
<td>Med-Pharmex</td>
<td>54925-063-10</td>
<td>&nbsp;</td>
<tr>
<td>Potassium acetate</td>
<td>&nbsp;</td>
<td>American Regent, Inc.</td>
<td>2053-25</td>
<td>&nbsp;</td>
<tr>
<td>Nitroglycerine</td>
<td>&nbsp;</td>
<td>American Regent, Inc.</td>
<td>4810-25 </td>
<td>&nbsp;</td>
<tr>
<td>Salix (Furosemide)</td>
<td>&nbsp;</td>
<td>Intervet Inc.</td>
<td>710461</td>
<td>&nbsp;</td>
<tr>
<td>5 Fr Hockey stick catheter</td>
<td>&nbsp;</td>
<td>Cordis</td>
<td>556-278-0L</td>
<td>&nbsp;</td>
<tr>
<td>7 Fr Hockey stick catheter</td>
<td>&nbsp;</td>
<td>Cordis</td>
<td>778-278-00</td>
<td>&nbsp;</td>
<tr>
<td>Coronary wire</td>
<td>&nbsp;</td>
<td>Abbott Laboratories</td>
<td>1044588</td>
<td>&nbsp;</td>
<tr>
<td>Coronary balloon</td>
<td>&nbsp;</td>
<td>Abbott Laboratories</td>
<td>1009447-08</td>
<td>&nbsp;</td>
<tr>
<td>8Fr sheath</td>
<td>&nbsp;</td>
<td>TERUMO</td>
<td>RSS801</td>
<td>&nbsp;</td>		</tbody>
		</table>
		</div>

gene transfer for ischemic heart failure in a preclinical model

다양한 새로운 기술은 심장 마비 환자에 대한 개발되고있다. 잘 설립 잠복기 평가는 효능과 안전성을 확인하기 위해 필요합니다. 바이러스성 벡터를 사용하여 유전자 치료는 심장 질환 치료를위한 가장 유망한 방법 중 하나입니다. 캐리어 유전자를 변경하여 여러 가지 유전자의 바이러스 배달 헤아릴 치료 가능성이있다. 이 비디오에서는 유전자 송금으로 다음 심장 마비 생성의 동물 모델의 전체 과정은 돼지 모델을 사용하여 제공됩니다. 첫째, 심근 경색은 관상 동맥을 내림차순 근위 왼쪽 앞쪽에을 occluding에 의해 생성됩니다. 만성 심장 마비에 심장 리모델링 결과. 우리의 모델 고유 정말 긍정적인 결과에 대한 공격적인 치료를 필요로 심각한 심장 마비 환자를 반영하는 상당히 큰 상처입니다. 후 심근 경색의 생성 및 반흔 조직의 개발, 바이러스의 intracoronary 주사는 동시 니트로 글리세린 주입과 증명합니다. 우리의 주입 방법은 향상된 유전자 발현과 함께 간단하고 효율적인 유전자 전달을 제공합니다. intracoronary 바이러스 전달과 함께 심근 경색 돼지 모델의 조합이 개별 유전자의 효과를 테스트하기 위해뿐만 아니라 도움이 일관성과 재현성 방법론,적일뿐만 아니라, 치료 후보로 많은 유전자의 효능을 비교했다.

Watch this Scientific Journal Video about Gene Transfer for Ischemic Heart Failure in a Preclinical Model at JoVE.com

Gene Transfer for Ischemic Heart Failure in a Preclinical Model

허혈성 심장 마비의 치료를위한 유전자 전달의 방법은 심근 경색의 돼지 모델을 사용하여 설명합니다. 우리 간단하고 재현성 방법은 우리가 쉽게 매우 간단하고 재현성 방법으로 다양한 유전자 전송의 효능을 평가할 수 있습니다.

잠복기 모델에서 허혈성 심장 마비에 대한 유전자 전송

Various emerging technologies are being developed for patients with heart failure. Well-established preclinical evaluations are necessary to determine ...

gene-transfer-for-ischemic-heart-failure-in-a-preclinical-model

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12.8K 조회수.  Mount Sinai School of Medicine. 허혈성 심장 마비의 치료를위한 유전자 전달의 방법은 심근 경색의 돼지 모델을 사용하여 설명합니다. 우리 간단하고 재현성 방법은 우리가 쉽게 매우 간단하고 재현성 방법으로 다양한 유전자 전송의 효능을 평가할 수 있습니다.

비디오: Gene Transfer for Ischemic Heart Failure in a Preclinical Model

A Simplified Technique for Producing an Ischemic Wound Model

One major obstacle in current diabetic wound research is a lack of an ischemic wound model that can be safely used in diabetic animals. Drugs that work well in non-ischemic wounds may not work in human diabetic wounds because vasculopathy is one major factor that hinders healing of these wounds. We published an article in 2007 describing a rabbit ear ischemic wound model created by a minimally invasive surgical technique. Since then, we have further simplified the procedure for easier operation. On one ear, three small skin incisions were made on the vascular pedicles, 1-2 cm from the ear base. The central artery was ligated and cut along with the nerve. The whole cranial bundle was cut and ligated, leaving only the caudal branch intact. A circumferential subcutaneous tunnel was made through the incisions, to cut subcutaneous tissues, muscles, nerves, and small vessels. The other ear was used as a non-ischemic control. Four wounds were made on the ventral side of each ear. This technique produces 4 ischemic wounds and 4 non-ischemic wounds in one animal for paired comparisons. After surgery, the ischemic ear was cool and cyanotic, and showed reduced movement and a lack of pulse in the ear artery. Skin temperature of the ischemic ear was 1-10 &deg;C lower than that on the normal ear and this difference was maintained for more than one month. Ear tissue high-energy phosphate contents were lower in the ischemic ear than the control ear. Wound healing times were longer in the ischemic ear than in the non-ischemic ear when the same treatment was used. The technique has now been used on more than 80 rabbits in which 23 were diabetic (diabetes time ranging from 2 weeks to 2 years). No single rabbit has developed any surgical complications such as bleeding, infection, or rupture in the skin incisions. The model has many advantages, such as little skin disruption, longer ischemic time, and higher success rate, when compared to many other models. It can be safely used in animals with reduced resistance, and can also be modified to meet different testing requirements.

We have developed a minimally invasive technique to create a rabbit ischemic ear wound model by dividing the central artery and nerve and the cranial neurovascular bundle. A subcutaneous tunnel then cuts all subcutaneous tissues. This procedure causes minimal skin disruption and can be safely used in diabetic animals.

허혈성 상처 모델 생산을위한 단순화된 테크닉

One major obstacle in current diabetic wound research is a lack of an ischemic wound model that can be safely used in diabetic animals. Drugs that work ...

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Permanent Ligation of the Left Anterior Descending Coronary Artery in Mice: A Model of Post-myocardial Infarction Remodelling and Heart Failure

Heart failure is a syndrome in which the heart fails to pump blood at a rate commensurate with cellular oxygen requirements at rest or during stress. It is characterized by fluid retention, shortness of breath, and fatigue, in particular on exertion. Heart failure is a growing public health problem, the leading cause of hospitalization, and a major cause of mortality. Ischemic heart disease is the main cause of heart failure.
Ventricular remodelling refers to changes in structure, size, and shape of the left ventricle. This architectural remodelling of the left ventricle is induced by injury (e.g., myocardial infarction), by pressure overload (e.g., systemic arterial hypertension or aortic stenosis), or by volume overload. Since ventricular remodelling affects wall stress, it has a profound impact on cardiac function and on the development of heart failure. A model of permanent ligation of the left anterior descending coronary artery in mice is used to investigate ventricular remodelling and cardiac function post-myocardial infarction. This model is fundamentally different in terms of objectives and pathophysiological relevance compared to the model of transient ligation of the left anterior descending coronary artery. In this latter model of ischemia/reperfusion injury, the initial extent of the infarct may be modulated by factors that affect myocardial salvage following reperfusion. In contrast, the infarct area at 24 hr after permanent ligation of the left anterior descending coronary artery is fixed. Cardiac function in this model will be affected by 1) the process of infarct expansion, infarct healing, and scar formation; and 2) the concomitant development of left ventricular dilatation, cardiac hypertrophy, and ventricular remodelling.
Besides the model of permanent ligation of the left anterior descending coronary artery, the technique of invasive hemodynamic measurements in mice is presented in detail.

Heart failure is the leading cause of hospitalization and a major cause of mortality. A model of permanent ligation of the left anterior descending coronary artery in mice is applied to investigate ventricular remodelling and cardiac dysfunction post-myocardial infarction. The technique of invasive hemodynamic measurements in mice is presented.

마우스의 왼쪽 전방 내림차순 관상 동맥의 영구 결찰 : 심근 경색 리모델링과 심장 마비의 모델

Heart failure is a syndrome in which the heart fails to pump blood at a rate commensurate with cellular oxygen requirements at rest or during stress. It ...

In Vivo Gene Transfer to the Rabbit Common Carotid Artery Endothelium

The goal of this method is to introduce a transgene into the endothelium of isolated segments of both rabbit common carotid arteries. The method achieves focal endothelial-selective transgenesis, thereby allowing an investigator to determine the biological roles of endothelial-expressed transgenes and to quantify the in vivo transcriptional activity of DNA sequences in large artery endothelial cells. The method uses surgical isolation of rabbit common carotid arteries and an arteriotomy to deliver a transgene-expressing viral vector into the arterial lumen. A short incubation period of the vector in the lumen, with subsequent aspiration of the lumen contents, is sufficient to achieve efficient and durable expression of the transgene in the endothelium, with no detectable transduction or expression outside of the isolated arterial segment. The method allows assessment of the biological activities of transgene products both in normal arteries and in models of human vascular disease, while avoiding systemic effects that could be caused either by targeting gene delivery to other sites (e.g. the liver) or by the alternative approach of delivering genetic constructs to the endothelium by germ line transgenesis. Application of the method is limited by the need for a skilled surgeon and anesthetist, a well-equipped operating room, the costs of purchasing and housing rabbits, and the need for expertise in gene-transfer vector construction and use. Results obtained with this method include: transgene-related alterations in arterial structure, cellularity, extracellular matrix, or vasomotor function; increases or reductions in arterial inflammation; alterations in vascular cell apoptosis; and progression, retardation, or regression of diseases such as intimal hyperplasia or atherosclerosis. The method also allows measurement of the ability of native and synthetic DNA regulatory sequences to alter transgene expression in endothelial cells, providing results that include: levels of transgene mRNA, levels of transgene protein, and levels of transgene enzymatic activity.

In Vivo Gene Transfer to the Rabbit Common Carotid Artery Endothelium

This method is to introduce a transgene into the endothelium of rabbit carotid arteries. Introduction of the transgene allows the assessment of the biological role of the transgene product either in normal arteries or disease models. The method is also useful for measuring activity of DNA regulatory sequences.

Vivo에서 토끼 일반적인 경 동맥 내 피에 유전자 이동

The goal of this method is to introduce a transgene into the endothelium of isolated segments of both rabbit common carotid arteries. The method achieves ...

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 ...

Isolation of Atrial Cardiomyocytes from a Rat Model of Metabolic Syndrome-related Heart Failure with Preserved Ejection Fraction

In this article, we describe an optimized, Langendorff-based procedure for the isolation of single-cell atrial cardiomyocytes (ACMs) from a rat model of metabolic syndrome (MetS)-related heart failure with preserved ejection fraction (HFpEF). The prevalence of MetS-related HFpEF is rising, and atrial cardiomyopathies associated with atrial remodeling and atrial fibrillation are clinically highly relevant as atrial remodeling is an independent predictor of mortality. Studies with isolated single-cell cardiomyocytes are frequently used to corroborate and complement in vivo findings. Circulatory vessel rarefication and interstitial tissue fibrosis pose a potentially limiting factor for the successful single-cell isolation of ACMs from animal models of this disease.
We have addressed this issue by employing a device capable of manually regulating the intraluminal pressure of cardiac cavities during the isolation procedure, substantially increasing the yield of morphologically and functionally intact ACMs. The acquired cells can be used in a variety of different experiments, such as cell culture and functional Calcium imaging (i.e., excitation-contraction-coupling).
We provide the researcher with a step-by-step protocol, a list of optimized solutions, thorough instructions to prepare the necessary equipment, and a comprehensive troubleshooting guide. While the initial implementation of the procedure might be rather difficult, a successful adaptation will allow the reader to perform state-of-the-art ACM isolations in a rat model of MetS-related HFpEF for a broad spectrum of experiments.

Here, we describe an optimized, Langendorff-based procedure for the isolation of single-cell atrial cardiomyocytes from a rat model of metabolic syndrome-related heart failure with preserved ejection fraction. A manual regulation of intraluminal pressure of cardiac cavities is implemented to yield functionally intact myocytes suitable for excitation-contraction-coupling studies.

심 방 Cardiomyocytes 보존된 방출 분수와 함께 대사 증후군 관련 심장 마비의 쥐 모델에서의 격리

In this article, we describe an optimized, Langendorff-based procedure for the isolation of single-cell atrial cardiomyocytes (ACMs) from a rat model of ...

Induction and Phenotyping of Acute Right Heart Failure in a Large Animal Model of Chronic Thromboembolic Pulmonary Hypertension

The development of acute right heart failure (ARHF) in the context of chronic pulmonary hypertension (PH) is associated with poor short-term outcomes. The morphological and functional phenotyping of the right ventricle is of particular importance in the context of hemodynamic compromise in patients with ARHF. Here, we describe a method to induce ARHF in a previously described large animal model of chronic PH, and to phenotype, dynamically, right ventricular function using the gold standard method (i.e., pressure-volume PV loops) and with a non-invasive clinically available method (i.e., echocardiography). Chronic PH is first induced in pigs by left pulmonary artery ligation and right lower lobe embolism with biological glue once a week for 5 weeks. After 16 weeks, ARHF is induced by successive volume loading using saline followed by iterative pulmonary embolism until the ratio of the systolic pulmonary pressure over systemic pressure reaches 0.9 or until the systolic systemic pressure decreases below 90 mmHg. Hemodynamics are restored with dobutamine infusion (from 2.5 &#181;g/kg/min to 7.5 &#181;g/kg/min). PV-loops and echocardiography are performed during each condition. Each condition requires around 40 minutes for induction, hemodynamic stabilization and data acquisition. Out of 9 animals, 2 died immediately after pulmonary embolism and 7 completed the protocol, which illustrates the learning curve of the model. The model induced a 3-fold increase in mean pulmonary artery pressure. The PV-loop analysis showed that ventriculo-arterial coupling was preserved after volume loading, decreased after acute pulmonary embolism and was restored with dobutamine. Echocardiographic acquisitions allowed to quantify right ventricular parameters of morphology and function with good quality. We identified right ventricular ischemic lesions in the model. The model can be used to compare different treatments or to validate non-invasive parameters of right ventricular morphology and function in the context of ARHF.

We present a protocol to induce and phenotype an acute right heart failure in a large animal model with chronic pulmonary hypertension. This model can be used to test therapeutic interventions, to develop right heart metrics&#160;or to improve the understanding of acute right heart failure pathophysiology.

만성 혈전색전성 폐 고혈압의 큰 동물 모델에서 급성 오른쪽 심부전의 유도 및 표고

The development of acute right heart failure (ARHF) in the context of chronic pulmonary hypertension (PH) is associated with poor short-term outcomes. The ...

Preclinical Model of Hind Limb Ischemia in Diabetic Rabbits

Peripheral vascular disease is a widespread clinical problem that affects millions of patients worldwide. A major consequence of peripheral vascular disease is the development of ischemia. In severe cases, patients can develop critical limb ischemia in which they experience constant pain and an increased risk of limb amputation. Current therapies for peripheral ischemia include bypass surgery or percutaneous interventions such as angioplasty with stenting or atherectomy to restore blood flow. However, these treatments often fail to the continued progression of vascular disease or restenosis or are contraindicated due to the overall poor health of the patient. A promising potential approach to treat peripheral ischemia involves the induction of therapeutic neovascularization to allow the patient to develop collateral vasculature. This newly formed network alleviates peripheral ischemia by restoring perfusion to the affected area. The most frequently employed preclinical model for peripheral ischemia utilizes the creation of hind limb ischemia in healthy rabbits through femoral artery ligation. In the past, however, there has been a strong disconnect between the success of preclinical studies and the failure of clinical trials regarding treatments for peripheral ischemia. Healthy animals typically have robust vascular regeneration in response to surgically induced ischemia, in contrast to the reduced vascularity and regeneration in patients with chronic peripheral ischemia. Here, we describe an optimized animal model for peripheral ischemia in rabbits that includes hyperlipidemia and diabetes. This model has reduced collateral formation and blood pressure recovery in comparison to a model with a higher cholesterol diet. Thus, the model may provide better correlation with human patients with compromised angiogenesis from the common co-morbidities that accompany peripheral vascular disease.

We describe a surgical procedure used to induce peripheral ischemia in rabbits with hyperlipidemia and diabetes. This surgery acts as a preclinical model for conditions experienced in peripheral artery disease in patients. Angiography is also described as a means to measure the extent of introduced ischemia and recovery of perfusion.

당뇨병 토끼에서의 뒷 사지의 전 임상 모델

Peripheral vascular disease is a widespread clinical problem that affects millions of patients worldwide. A major consequence of peripheral vascular ...

Establishing a Swine Model of Post-myocardial Infarction Heart Failure for Stem Cell Treatment

Although advances have been achieved in the treatment of heart failure (HF) following myocardial infarction (MI), HF following MI remains one of the major causes of mortality and morbidity around the world. Cell-based therapies for cardiac repair and improvement of left ventricular function after MI have attracted considerable attention. Accordingly, the safety and efficacy of these cell transplantations should be tested in a preclinical large animal model of HF prior to clinical use. Pigs are widely used for cardiovascular disease research due to their similarity to humans in terms of heart size and coronary anatomy. Therefore, we sought to present an effective protocol for the establishment of a porcine chronic HF model using closed-chest coronary balloon occlusion of the left circumflex artery (LCX), followed by rapid ventricular pacing induced with pacemaker implantation. Eight weeks later, the stem cells were administered by intramyocardial injection in the peri-infarct area. Then the infarct size, cell survival, and left ventricular function (including echocardiography, hemodynamic parameters, and electrophysiology) were evaluated. This study helps establish a stable preclinical large animal HF model for stem cell treatment.

We sought to establish a swine model of heart failure induced by left circumflex artery blockage and rapid pacing to test the effect and safety of intramyocardial administration of stem cells for cell-based therapies.

줄기 세포 치료를위한 심근 경색 심부전의 돼지 모델 설립

Although advances have been achieved in the treatment of heart failure (HF) following myocardial infarction (MI), HF following MI remains one of the major ...

A Preclinical Model of Exertional Heat Stroke in Mice

Heat stroke is the most severe manifestation of heat-related illnesses. Classic heat stroke (CHS), also known as passive heat stroke, occurs at rest, whereas exertional heat stroke (EHS) occurs during physical activity. EHS differs from CHS in etiology, clinical presentation, and sequelae of multi-organ dysfunction. Until recently, only models of CHS have been well established. This protocol aims to provide guidelines for a refined preclinical mouse model of EHS that is free from major limiting factors such as the use of anesthesia, restraint, rectal probes, or electric shock. Male and female C57Bl/6 mice, instrumented with core temperature (Tc) telemetric probes were utilized in this model. For familiarization with the running mode, mice undergo 3 weeks of training using both voluntary and forced running wheels. Thereafter, mice run on a forced wheel inside a climatic chamber set at 37.5 &#176;C and 40%-50% relative humidity (RH) until displaying symptom limitation (e.g., loss of consciousness) at Tc of 42.1-42.5 &#176;C, although suitable results can be obtained at chamber temperatures between 34.5-39.5 &#176;C and humidity between 30%-90%. Depending on the desired severity, mice are removed from the chamber immediately for recovery in ambient temperature or remain in the heated chamber for a longer duration, inducing a more severe exposure and a higher incidence of mortality. Results are compared with sham-matched exercise controls (EXC) and/or na&#239;ve controls (NC). The model mirrors many of the pathophysiological outcomes observed in human EHS, including loss of consciousness, severe hyperthermia, multi-organ damage as well as inflammatory cytokine release, and acute phase responses of the immune system. This model is ideal for hypothesis-driven research to test preventative and therapeutic strategies that may delay the onset of EHS or reduce the multi-organ damage that characterizes this manifestation.

The protocol describes the development of a standardized, repeatable, preclinical model of exertional heat stroke (EHS) in mice free from adverse external stimuli such as electric shock. The model provides a platform for mechanistic, preventative, and therapeutic studies.

마우스에서 운동 열사병의 전임상 모델

Heat stroke is the most severe manifestation of heat-related illnesses. Classic heat stroke (CHS), also known as passive heat stroke, occurs at rest, ...

Large Animal Model for Evaluating the Efficacy of the Gene Therapy in Ischemic Heart

Coronary artery disease is one of the significant causes of mortality and morbidity worldwide. Despite the progression of current therapeutics, a considerable proportion of coronary artery disease patients remain symptomatic. Gene therapy-mediated therapeutic angiogenesis offers a novel therapeutic method for improving myocardial perfusion and relieving symptoms. Gene therapy with different angiogenic factors has been studied in few clinical trials. Due to the novelty of the method, the progress of myocardial gene therapy is a continuous path from bench to bedside. Therefore, large animal models are needed for evaluating the safety and efficacy. The more the large animal model identifies the original disease and the endpoints used in clinics, the more predictable outcomes are from clinical trials. Here, we introduce a large animal model for evaluating the efficacy of the gene therapy in the ischemic porcine heart. We use clinically relevant imaging methods such as ultrasound imaging and 15H2O-PET. For targeting the gene transfers into the desired area, electroanatomical mapping is used. The aim of this method is: (1) to mimic chronic coronary artery disease, (2) to induce therapeutic angiogenesis at hypoxic areas of the heart, and (3) to evaluate the safety and efficacy of the gene therapy by using relevant endpoints.

Myocardial gene therapy for ischemic heart disease holds great promise for future therapeutics. Here, we introduce a large animal model for evaluating the efficacy of gene therapy in the ischemic heart.