Name: 3d whole-heart myocardial tissue analysis
Uploaded: 2017-04-12T13:00:00
Description: Watch this Scientific Journal Video about 3D Whole-heart Myocardial Tissue Analysis at JoVE.com

笔者想感谢马利贾恩·詹森，霍伊塞·葳瑟和马亭面包车Nieuwburg他们与动物实验的援助。我们非常感谢的Martijn Froeling和安克·沃辛克他们与MRI成像援助。

体内实验按照指南的护理和使用的实验动物研究所的准备实验动物进行。实验批准由当地动物实验委员会。 1.可注射的和嵌入溶液的制备<ol><li>准备注射的凝胶。 <ol><li>按照制备1毫升脲基-嘧啶酮（UPy）凝胶的到先前描述的方案12,13。 </li><li>添加超顺磁性氧化铁颗粒（SPIOs）到该溶液中以获得15微克/ mL的浓度，并搅拌5分钟?...

<ol>
	<li>Nowbar, A. N., Howard, J. P., Ja Finegold, ., Asaria, P., Francis, D. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=2014+global+geographic+analysis+of+mortality+from+ischaemic+heart+disease+by+country,+age+and+income:+statistics+from+World+Health+Organisation+and+United+Nations.">2014 global geographic analysis of mortality from ischaemic heart disease by country, age and income: statistics from World Health Organisation and United Nations.</a> Int J Cardiol. 174 (2), 293-298 (2014).</li><li>Kannel, W. B., Belanger, A. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Epidemiology+of+heart+failure.">Epidemiology of heart failure.</a> Am Heart J. 121 (3), 951-957 (1991).</li><li>Ib&#225;&#241;ez, B., Heusch, G., Ovize, M., Van De Werf, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evolving+therapies+for+myocardial+ischemia/reperfusion+injury.">Evolving therapies for myocardial ischemia/reperfusion injury.</a> J Am Coll Cardiol. 65 (14), 1454-1471 (2015).</li><li>Bartunek, J., Vanderheyden, M., Hill, J., Terzic, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cells+as+biologics+for+cardiac+repair+in+ischaemic+heart+failure.">Cells as biologics for cardiac repair in ischaemic heart failure.</a> Heart. 96 (10), 792-800 (2010).</li><li>Orlic, D., Kajstura, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bone+marrow+cells+regenerate+infarcted+myocardium.">Bone marrow cells regenerate infarcted myocardium.</a> Nature. 410 (6829), 701-705 (2001).</li><li>Nguyen, P. K., Lan, F., Wang, Y., Wu, J. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Imaging:+Guiding+the+Clinical+Translation+of+Cardiac+Stem+Cell+Therapy.">Imaging: Guiding the Clinical Translation of Cardiac Stem Cell Therapy.</a> Circ Res. 109 (8), 962-979 (2011).</li><li>Psaltis, P. J., Worthley, S. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Endoventricular+electromechanical+mapping-the+diagnostic+and+therapeutic+utility+of+the+NOGA+XP+Cardiac+Navigation+System.">Endoventricular electromechanical mapping-the diagnostic and therapeutic utility of the NOGA XP Cardiac Navigation System.</a> J Cardiovasc Transl Res. 2 (1), 48-62 (2009).</li><li>Tomkowiak, M. T., Klein, A. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Targeted+transendocardial+therapeutic+delivery+guided+by+MRI-x-ray+image+fusion.">Targeted transendocardial therapeutic delivery guided by MRI-x-ray image fusion.</a> Catheter Cardiovasc Interv. 78 (3), 468-478 (2011).</li><li>Dauwe, D. F., Nuyens, D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Three-dimensional+rotational+angiography+fused+with+multimodal+imaging+modalities+for+targeted+endomyocardial+injections+in+the+ischaemic+heart.">Three-dimensional rotational angiography fused with multimodal imaging modalities for targeted endomyocardial injections in the ischaemic heart.</a> Eur Heart J Cardiovasc Imaging. 15 (8), 900-907 (2014).</li><li>van Slochteren, F. J., van Es, R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Multimodality+infarct+identification+for+optimal+image-guided+intramyocardial+cell+injections.">Multimodality infarct identification for optimal image-guided intramyocardial cell injections.</a> Neth Heart J. 22 (11), 493-500 (2014).</li><li>van Slochteren, F. J., van Es, R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Three+dimensional+fusion+of+electromechanical+mapping+and+magnetic+resonance+imaging+for+real-time+navigation+of+intramyocardial+cell+injections+in+a+porcine+model+of+chronic+myocardial+infarction.">Three dimensional fusion of electromechanical mapping and magnetic resonance imaging for real-time navigation of intramyocardial cell injections in a porcine model of chronic myocardial infarction.</a> Int J Cardiovasc Imaging. 32 (5), 833-843 (2016).</li><li>Pape, a. C. H., Bakker, M. H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+Injectable+and+Drug-loaded+Supramolecular+Hydrogel+for+Local+Catheter+Injection+into+the+Pig+Heart.">An Injectable and Drug-loaded Supramolecular Hydrogel for Local Catheter Injection into the Pig Heart.</a> J Vis Exp. (100), (2015).</li><li>Bastings, M. M. C., Koudstaal, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+fast+pH-switchable+and+self-healing+supramolecular+hydrogel+carrier+for+guided,+local+catheter+injection+in+the+infarcted+myocardium.">A fast pH-switchable and self-healing supramolecular hydrogel carrier for guided, local catheter injection in the infarcted myocardium.</a> Adv Healthc Mater. 3 (1), 70-78 (2014).</li><li>Koudstaal, S., Jansen of Lorkeers, S. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Myocardial+infarction+and+functional+outcome+assessment+in+pigs.">Myocardial infarction and functional outcome assessment in pigs.</a> J. Vis. Exp. (86), (2014).</li></ol>

全心脏根据该协议3D心肌组织处理提供能够使梗塞，所述IBZ，并相对于心脏解剖结构的执行的注射的三维分析的结构化方法。心脏的灌装量取决于所需的分析。在这项研究中，以评估注射的准确性，我们的目的是填补心脏，以尽可能地类似于舒张末期的几何形状。为了执行此，左心室尖被固定到所述容器的底部和LV充满琼脂而肺静脉被夹紧。当LV被填充，主动脉被夹紧以及，防止琼脂流出LV和作为?...

缺血性心脏疾病一直是世界领先的死亡原因为过去几十年中1。心肌梗死后急性治疗的目的是通过经皮冠状动脉介入或冠状动脉旁路移植术恢复血流到心肌。在严重的脑梗塞，心肌大面积是伤痕累累，而这些情况往往导致缺血性心脏衰竭（HF）2。为HF注重预防和心脏功能的保存对于心衰患者目前的治疗方案，但不是再生。 在过去的十年中，心脏再生疗法已被研究作为HF 3的一种治疗选择。此治疗的目的是提供生物制剂，例如干细胞或生长因子，直接将损伤心肌，以诱导血管再生，心肌保护，分化和生长4。为了获得最佳的治疗效果，据推测，生物必须在梗死边缘区（IBZ）被喷射到促进良好的组织灌注的生物的生存和用于向所述目标区5，6最佳的效果。多种技术已被开发来执行IBZ的识别和可视化，以引导心肌内注射7，8，9，10，11。除了IBZ的识别和可视化，所述输送也依赖于所使用的生物材料和注射导管。为了验证的递送技术的喷射精度，需要精确的和可重现的定量方法。 我们已经开发了能够提供二维（2D）的全心脏心肌组织处理和三dimensio协议纳尔（3D）成像，其可用于定性和定量研究的目的。该协议覆盖的嵌入过程和数字图像分析。在本文中，我们证明了在心肌内IBZ注射的定位的准确性慢性心肌梗死的大猪模型评估的协议。

<table border="0" cellpadding="0" cellspacing="0" width="610">
	<tbody>
		<tr height="20">
			<td height="20" style="height:20px;width:216px;">0.9% Saline</td>
			<td style="width:108px;">Braun</td>
			<td style="width:119px;"></td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="38">
			<td height="38" style="height:38px;width:216px;">Agarose</td>
			<td style="width:108px;">Roche Diagnostics</td>
			<td style="width:119px;"></td>
			<td style="width:167px;">Scientific grade multipurpose agar</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Biomolecular fluorescence scanner Typhoon 9410&#160;</td>
			<td>GE Healthcare</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:216px;">Embedding container</td>
			<td style="width:108px;"></td>
			<td style="width:119px;"></td>
			<td>Plastic, dimensions 17 x 14,5 x 14 cm</td>
		</tr>
		<tr height="39">
			<td height="39" style="height:39px;width:216px;">FluoSpheres Polystyrene Microspheres</td>
			<td style="width:108px;">Invitrogen</td>
			<td>F8834</td>
			<td>red, 10 &#181;m</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:216px;">Gadolinium</td>
			<td style="width:108px;">Gadovist</td>
			<td style="width:119px;"></td>
			<td>1,0 mmol/mL</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:216px;">dS 32 channel head coil</td>
			<td style="width:108px;">Philips</td>
			<td style="width:119px;"></td>
			<td>Or similar</td>
		</tr>
		<tr height="38">
			<td height="38" style="height:38px;width:216px;">Matlab</td>
			<td style="width:108px;">Mathworks</td>
			<td style="width:119px;"></td>
			<td style="width:167px;">To insure compatability 2015a or newer</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:216px;">Meat slicer</td>
			<td style="width:108px;">Berkel</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="39">
			<td height="39" style="height:39px;width:216px;">Myostar injection catheter</td>
			<td style="width:108px;">Biosense Webster</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="39">
			<td height="39" style="height:39px;width:216px;">Super paramagnetic iron oxide particles</td>
			<td style="width:108px;">Sinerem</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:216px;">Triphenyl-tetrazolium chloride</td>
			<td style="width:108px;">Merck</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">UPy-PEG10k</td>
			<td style="width:108px;"></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:216px;">Vicryl 2-0</td>
			<td style="width:108px;">Ethicon</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
	</tbody>
</table>

3d whole-heart myocardial tissue analysis

心脏再生疗法的目的是保护和治疗缺血性心脏疾病修复受伤的心脏。通过注入的干细胞或增强血管造影或血管入梗死边缘区（IBZ）其他生物，组织灌注得到改善，并且心肌可以免受进一步损伤的保护。为了获得最大的治疗效果，这是假设，再生剂最好交付给IBZ。这需要精确的注射，并导致新的注射技术的发展。为了验证这些新的技术，我们已经基于心肌组织分析设计验证协议。这个协议包括全心脏心肌组织处理，使详细的二维（2D）和心脏解剖和心肌内注射的三维（3D）分析。在猪，心肌梗死是由左前降冠状动脉的90分钟闭塞创建。 4周后，一个mixt超顺磁性氧化铁颗粒（SPIOs）和荧光珠的水凝胶的URE用微创心内膜的方法在IBZ注入。在注射过程后1个小时，猪安乐死，和心脏切下并包埋在琼脂糖（琼脂）。琼脂凝固后，进行磁共振成像（MRI），心脏的切片，和荧光成像。图像后处理后，进行三维分析，以评估IBZ定位精度。这个协议提供了心肌内注射的靶向精度进IBZ的评估结构化和可再现的方法。该协议可以期望瘢痕组织和/或整个心脏的喷射精度的验证的处理时，可以容易地使用。

组织包埋通过嵌入过程，舒张末期般的几何成立。成功地附着于心脏组织，使组织中的琼脂上以在与等于切片厚度（ 图2A和2C）的所需角度进行切片。 Scar-和注射部位评估对于每个成像模态，梗塞和喷射位置的评估...

Watch this Scientific Journal Video about 3D Whole-heart Myocardial Tissue Analysis at JoVE.com

3D Whole-heart Myocardial Tissue Analysis

这个协议描述了用于与MRI全心脏心肌组织的3D比较的新方法。这是专为心肌内注射的心肌梗死的慢性猪模型的梗死边缘区的准确评估。

3D全心脏心肌组织分析

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

Research

JoVE Journal

Medicine

Acute Myocardial Infarction in Rats

大鼠急性心肌梗死

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

科研

医学

A Research Method For Detecting Transient Myocardial Ischemia In Patients With Suspected Acute Coronary Syndrome Using Continuous ST-segment Analysis

研究方法检测疑似急性冠脉综合征患者短暂心肌缺血的连续ST段分析

Each year, an estimated 785,000 Americans will have a new coronary attack, or acute coronary syndrome (ACS). The pathophysiology of ACS involves rupture ...

Ultrasonic Assessment of Myocardial Microstructure

心肌微观结构超声波评估

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

Myocardial Infarction and Functional Outcome Assessment in Pigs

心肌梗死和功能预后评估的猪

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

Histological Quantification of Chronic Myocardial Infarct in Rats

大鼠慢性心肌梗塞的组织学定量分析

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

3D Ultrasound Imaging: Fast and Cost-effective Morphometry of Musculoskeletal Tissue

3D 超声成像: 快速、经济高效的肌骨骼组织形态分析

The developmental goal of 3D ultrasound imaging (3DUS) is to engineer a modality to perform 3D morphological ultrasound analysis of human muscles. 3DUS ...

Improvement of a Closed Chest Porcine Myocardial Infarction Model by Standardization of Tissue and Blood Sampling Procedures

组织和采血程序标准化改善闭合性胸猪心肌梗死模型的研究

Myocardial ischemia reperfusion (I/R) injury contributes to almost half of the necrotic area after myocardial infarction. To date there is no approved ...

Real-time Pressure-volume Analysis of Acute Myocardial Infarction in Mice

小鼠急性心肌梗死的实时压力容积分析

Acute myocardial infarction can lead to acute heart failure and cardiogenic shock. The evaluation of hemodynamics is critical for the evaluation of any ...

Generation of Human 3D Lung Tissue Cultures (3D-LTCs) for Disease Modeling

Generation of Human 3D Lung Tissue Cultures 3D-LTCs for Disease Modeling

用于疾病建模的人三维肺组织培养 (3D-LTCs) 的生成

Translation of novel discoveries to human disease is limited by the availability of human tissue-based models of disease. Precision-cut lung slices (PCLS) ...

Use of 3D Robotic Ultrasound for In Vivo Analysis of Mouse Kidneys

Use of 3D Robotic Ultrasound for In Vivo Analysis of Mouse Kidneys

使用3D机器人超 声进行小鼠 肾脏体内分析

Common modalities for in vivo imaging of rodents include positron emission tomography (PET), computed tomography (CT), magnetic resonance imaging (MRI), ...