作者感谢克里斯蒂安Pahrmann她的技术援助。这项研究是由德国妇儿基金会资助Herzforschung（F / 28/14）。 DW是由来自国际社会旅游奖心脏和肺移植的支持。 TD从否则-克朗 - 费森尤斯基金会（2012_EKES.04）收到否则克朗卓越助学金。 SS收到研究经费来自德意志研究联合会（DFG; DE2133 / 2-1，TD和SCHR992 / 3- 1，SCHR992 / 4-1，SS）。

动物遵照指南实验动物的原则，通过实验动物资源研究所编制和美国国立卫生研究院公布收到的人文关怀。所有动物方案是由主管地方当局（"献给金额UND GESUNDHEIT Verbraucherschutz，Hansestadt（办公室健康与消费者保护）汉堡"）的批准。 1.动物护理<ol><li>获取Lewis大鼠（LEW / CRL）大鼠和ROSA /荧光素酶LEW转基因大鼠实验动物研究所体重300-350克</li><li>保持常规条件下的老鼠在通风柜和养活他们的标准大鼠饲料和蒸压水随意。 </li><li>执行使用ROSA /荧光素酶LEW转基因大鼠作为捐助者和同系LEW / CRL大鼠作为收件人移植术。 </li></ol> 2.捐助者大鼠的制备<ol><li>使用吸纳离子腔麻醉异氟烷（2.5-3％）的老鼠。 </li><li>放置在它的后面的大鼠和保持与覆盖嘴和鼻子的面罩的麻醉。按捏后脚和验证的情况下反射的检查麻醉足够的深度。涂一些药膏兽医的眼睛，以防止干燥，而在麻醉下。 </li><li>传播后腿和用胶布固定自己的位置。 </li><li>剃腹股沟头发与头发剪，并使用聚维酮碘，随后通过80％乙醇消毒的整个区域。重复消毒一步的两倍。 注:手术区，纱布，及手术器械应灭菌。维持无菌区整个过程和穿一次性使用的，无菌手术手套，口罩，和帽。 </li><li>在显微镜下，进行沿LINEA inguinalis垂直切口。用两个镊子皮下组织轻轻分开，并从其ORI暴露腹壁浅静脉杜松子酒的股静脉。小心地从周围组织中分离出腹壁浅静脉。 </li><li>停止使用两个微夹子固定腹壁浅静脉血流。 </li><li>通过仔细地抬起与镊子分离静脉，并通过容器与microscissors切割收获静脉的大致0.5到1厘米的段。留在容器残端微夹具防止血液流失。放置在无菌纱布取下的一块静脉。小心地将一地下30针收获静脉的一端内侧和冲洗用肝素容器（50单位/ ml）。 注意:小心使用静脉，避免吊装，切割，和冲洗过程中的损坏。确保肝素适量的冲洗移植。 </li><li>保持在冰上在1％利多卡因的血管区段，直到移植到受体鼠，以防止一个容器痉挛。 </li><li>通过增加麻醉5％异氟醚安乐死供体鼠。后2-3分钟，运恩沿着白线腹部，划破膜片，并取出心脏停止流通。 </li></ol> 3.收件人鼠的制备<ol><li>麻醉并固定收件人大鼠以相同的方式作为供体鼠。 </li><li>剃腿的内侧与毛发修剪器，并使用聚维酮碘和80％乙醇消毒三次。 </li><li>监测麻醉深度，并确保它是按捏后脚时验证的情况下的反射的足够了。 </li><li>从膝盖到腹股沟褶执行中值股骨切口。在显微镜下，用2镊子股动脉从周围环境中分离出来。 </li><li>使用微夹钳阻止血液的流动。放置近侧夹紧第一，其次远侧夹子。 </li><li>剪出夹紧股动脉microscissors短段和丢弃。缩短剩下的树桩动脉与microscissors，创造一个差距是1-2毫米比静脉移植大。利用地下30针冲洗动脉残端与肝素。 注:如果外膜伸出略微超出血管断端，使用镊子小幅拉过来的船只将其取出一块。 </li><li>从步骤2.8动脉树桩之间放置收集静脉和调整长度，使其合适地配合到间隙。注静脉的方向。 </li><li>首先用一个10-0普理灵缝合执行近端吻合。在所示（ 图1D）的顺序进行单针。开始与腹侧增加三个缝合线之前的每个侧面缝合。之后，放置在容器的背侧三针来完成吻合术。 </li><li>与使用相同的技术，作为在步骤3.8中描述的近端吻合接枝连接远侧血管。再次，开始在每个横向侧的缝合线，然后将三道缝线上的腹侧的side和背侧。 </li><li>负载两个1毫升注射器用纤维蛋白胶成分1和2小心提起用镊子接枝拖放大约100μl的纤维蛋白胶部件1的接枝下，接着部件2。 注:确保组件1和2在1应用:1的比例。 </li><li>放置接枝背部在其位置，并在移植物的顶部下降一个额外的100μl的组分1和2的。可以肯定的胶水涵盖接枝并且为了防止吻合不全和静脉移植物的过度膨胀的吻合术。 </li><li>小心地打开远端夹紧，随后近端。 </li><li>确认通过检查在移植静脉和移植物的远端动脉可见光脉冲一个成功的手术。 </li><li>除去过多的胶水，阻碍皮肤缝合。使用镊子解除固化胶水和microscissors去除多余的。关闭皮肤层用5-0普理灵缝线。 </li><li>注射4-5米克/公斤卡洛芬皮下注射使鼠醒来了。不要离开无人看管的动物，直到它已经恢复了足够的意识，以保持胸骨斜卧。保持动物在一个单一的笼子里，直到它完全恢复。 </li><li>添加安乃近到饮用水（每100毫升50毫克安乃近）作为止痛药为以下3天，每日监测动物。 </li></ol> 4.多普勒超声注:使用超声duxplex可视化血流非侵入性大鼠14。 <ol><li>麻醉中的感应腔（异氟烷2％）大鼠。将在它的后面大鼠和与覆盖鼻面罩维持麻醉。 </li><li>使用电推剪及脱毛霜去除周围的大腿区域的头发。 </li><li>应用超声波凝胶大腿。确保有没有气泡。使用MS 400传感器获取多普勒超声图像（中心频率:30男赫兹）与230-400帧/秒的帧速率。 </li></ol> 5.组织病理学注:嘉实和染色与马松染色的容器形态分析15。 <ol><li>过夜固定在4％低聚甲醛的收获容器和在增加的乙醇浓度脱水它。嵌入石蜡的样品，并将其切成用切片机5微米厚的切片。 注:多聚甲醛是有毒的，应特别小心。 </li><li>用三色染色液染色之前Deparaffinize的幻灯片。脱水染色切片，用二甲苯清除它们，在安装中安装它们。干燥载玻片后，用显微镜观看样本。 </li></ol> 6.生物发光成像（BLI） 注:移植术后通过测定生物发光信号1跟踪随着时间的推移在体内6。 <ol><li>溶解1g D-荧光素钾盐在22ml的PBS中，并将其注入腹膜内到大鼠（375毫克/千克体重）。等待15分钟的荧光素在动物中循环。 </li><li>将老鼠变成一个实时定量生物发光系统并访问生物发光信号。 </li></ol>

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The authors have nothing to disclose.

本视频演示了大鼠静脉插入模型，探讨静脉移植的损失，并允许潜在的病理过程的探索和新的药物和治疗方案的测试。 麻醉的手术过程中的一个关键方面。建议连续吸入性麻醉系统，因为这是一种安全，简便的方法，尤其是在长时间的操作。这可以在训练阶段期间，当操作的时间超过1小时，是非常重要的。 相对于手术过程，重要的是不向收获及植入17时损坏静脉移植物。夹持容器的外膜可以防止对静脉移植损失，避免随后的血栓形成和移植物衰竭。移植通畅可以立即在静脉移植物或通过DISTA血液流动的观察与脉动移植静脉建设后确定升动脉，以及通过双功能超声。 该方法的另一个重要方面是，以防止静脉移植物的过度扩张。静脉移植到动脉压系统突然暴露导致增加壁张力，随后的过度扩张，并在流动格局的变化。这些都是血栓形成，吻合口机能不全，早期移植失败的源。支持与纤维蛋白胶移植静脉可以防止不受控制的气球和防止机械破坏内膜和媒体。吸收的胶原封面是纤维蛋白胶的替代和可以用作静脉周围覆盖18。 该协议中最关键的一步，无疑是静脉移植物和动脉之间的吻合。必须小心不要刺穿容器的两个壁，因为这会导致吻合的缩小，这可导致移植物的早期失效。另外，特别注意要n必须支付给缝线的本地化。在动脉和静脉缝合位置的一致性，确保移植通畅，防止不足。为方便起见，缝线可以在图1D中所示的顺序来执行。 技术的难易程度可以被看作是该技术的限制，因为新手必须首先熟悉显微设备和所述解剖结构的小尺寸。然而，使用其他车型表现出相同的困难，我们相信，该视频将帮助外科医生新手掌握在短时间内这种技术。 无数小动物模型静脉移植失败已经在文献中进行了描述。然而，大多数车型只分析13提供了非常少量组织。这种方法的一个优点是，可以得到相当大量的组织。一种接枝可以被划分成多个部分的次用于不同测定法，从而减少了所需的实验动物的数量。 在锌指核酸酶技术的最新进展使敲除鼠19的产生。通过选择合适的基因敲除大鼠移植通畅的损失可以在不同的病情进行研究。例如，肾素敲除鼠可用于研究高血压20。这些遗传背景可以用此动物模型相结合，以收集关于静脉移植失败的在不同的设置或对某些基因在myointima增生的发展的影响的机制的信息。 总之，在此视频描述的模型是可重复的，价格低廉，且容易执行，并且它可以快速而可靠地建立。 Myointima增生，这是静脉移植失败的主要原因，迅速发展四个星期，造成渐进腔闭塞。试验成功治疗在这个模型选项可以在大动物模型中进一步证实。

冠状动脉疾病及其并发症是世界范围内死亡的主要原因之一。当前的治疗策略集中在重新建立血流，或者通过扩张变窄容器或通过创建旁路。使用自体静脉冠状动脉旁路移植术（CABG）于1968年首次描述，并不断得到完善，多年来。除了左前的血运重建降支动脉，隐静脉导管是最常用的1。然而，移植通畅仍然隐静脉移植（SVG）的致命弱点。手术一年后，通畅率85％，下降幅度为61％，十年后2,3。因此揭幕SVG通畅损失的病理生理机制和原因的一项重要任务。 本视频演示了大鼠静脉插入模型，探讨静脉移植物损失。这种方法的总体目标是发掘潜在的病理生物学并在疾病进展和-physiological流程，制定药物或治疗方法测试一个合适的模型。通过移植的腹壁浅静脉进入动脉系统，这种模式密切模仿冠状动脉旁路移植术的临床设置。手术创伤，局部缺血，和壁应力是病理血管变化重要触发器和在所述的模型模仿。 不同型号和种类都可以探讨静脉移植通畅的损失。大动物模型，如猪4，羊5，狗6和猴7，类似于人类血管和解剖结构，从而使复杂的治疗策略，如旁路支架或新的外科技术，待测试8。然而，需要特殊的房屋，设备，人员和工作人员。此外，高的成本和手术过程中需要额外的麻醉师阻碍其更广泛的应用。钐所有的动物，包括老鼠，很容易处理，不需要特殊的住房，并有管理的成本。相比于小鼠模型9,10，大鼠模型具有较好的可操作性的成果优势，从而减少可变性。老鼠是生理和基因更类似于人类比老鼠11,12。此外，大多数的野生型小鼠仅开发有限myointima 13，其使小鼠模型容易发生II型误差。主小鼠静脉，如下腔静脉的组织学，只由几个细胞层的并呈现早期评估困难13。另一个缺点是可用于接枝恢复之后的后续分析的组织的量小。 在此视频描述的模型是可重复的，价格低廉，且容易执行，并且它可以快速而可靠地建立。它特别适合用于评估昂贵的实验治疗剂，如病毒载体用于基因疗法，以经济的方式。

<table border="0" cellpadding="0" cellspacing="0" width="720">
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			<td height="51" style="height:51px;width:236px;">Rat LEW-Tg(Gt(ROSA)26Sor- 1 
			luc)11Jmsk</td>
			<td style="width:183px;">Institute of laboratory animals, Kyoto University, Japan</td>
			<td style="width:145px;">NBPR rat number 0299</td>
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			<td height="17" style="height:17px;">Novaminsulfon</td>
			<td>Ratiopharm</td>
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			<td height="17" style="height:17px;">Heparin</td>
			<td>Rotexmedica</td>
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			<td height="17" style="height:17px;">Xylocain 1%</td>
			<td>AstraZeneca</td>
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			<td height="38" style="height:38px;">EVICEL</td>
			<td>J&#38;J Med.Ethicon Biosur</td>
			<td style="width:145px;">PZN 7349697 Art. Nr.:EVK01DE</td>
			<td>fibrin glue</td>
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			<td height="37" style="height:37px;">NaCl 0,9%</td>
			<td>B.Braun</td>
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			<td height="34" style="height:34px;width:236px;">Vevo 770 high-resolution in vivo micro-imaging system</td>
			<td>VisualSonics</td>
			<td></td>
			<td>duplex sonography</td>
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			<td height="17" style="height:17px;">Ecogel 100 ultrasound gel</td>
			<td>Eco-med</td>
			<td>30GB</td>
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			<td>Perkin Elmer</td>
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			<td>bioluminescence imaging</td>
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			<td height="17" style="height:17px;">Weigerts iron hematoxylin Kit</td>
			<td>Merck</td>
			<td>1.15973.0002</td>
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			<td height="17" style="height:17px;">Acid Fuchsin</td>
			<td>Sigma-Aldrich</td>
			<td>F8129-25G</td>
			<td>Trichrome staining</td>
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			<td height="17" style="height:17px;">Ponceau S solution</td>
			<td>Serva Electrophoresis</td>
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			<td height="17" style="height:17px;">Paraffin</td>
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			<td height="17" style="height:17px;">Ethanol 70%</td>
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</table>

vein interposition model: a suitable model to study bypass graft patency

Bypass grafting is an established treatment method for coronary artery disease. Graft patency continues to be the Achilles heel of saphenous vein grafts. Research models for bypass graft failure are essential for a better understanding of pathobiological and pathophysiological processes during graft patency loss. Large animal models, such as pigs or sheep, resemble human anatomical structures but require special facilities and equipment. This video describes a rat vein interposition model to investigate vein graft patency loss. Rats are inexpensive and easy to handle. Compared to mouse models, the convenient size of rats permits better operability and enables a sufficient amount of material to be obtained for further diverse analysis. In brief, the inferior epigastric vein of a donor rat is harvested and used to replace a segment of the femoral artery. Anastomosis is conducted via single stitches and sealed with fibrin glue. Graft patency can be monitored non-invasively using duplex sonography. Myointimal hyperplasia, which is the main cause for graft patency loss, develops progressively over time and can be calculated from histological cross sections.

大鼠静脉插入模型适用于研究myointima增生静脉移植失败的发展。动物从手术中恢复良好，显示出出色的身体条件后的操作。 图1示出了关键的手术步骤。沿着LINEA inguinalis切开皮肤后，上腹部浅静脉和股动脉被确定（ 图1A）。应仔细进行接枝的收获，而不会损坏接枝（ 图1B），因为这可能导致早期移植物衰竭和通畅损失。在受体动物（ 图1C）定位接枝后，吻合针迹在图1D中所示的顺序执行。完成后的静脉移植介入出现面色苍白（ 图1E），并应成为红色，并显示灌注（ 图1F）后的脉动。 成功的集成公司<html>在静脉进入股动脉和移植物通畅的移植后可以确认非侵入性使用双面超声（ 图2A）。通过移植一个吕克阳性大鼠的静脉进入一个同基因吕克负鼠，生物发光成像可以被用于监测在一段时间（ 图2B）接枝的存在。 Myointima增生移植随着时间的推移逐步发展起来的。用Masson三色组织染色表明内弹性膜（ 图2C）内myointima形成。管腔闭塞的计算，（分割管腔的截面积由区域内弹性膜内）显示移植物通畅的从第7天至28天手术后逐渐损失，从而确认myointima增生的再生动态在此大鼠模型中（ 图2D）。 重1"SRC ="/文件/ ftp_upload / 54839 / 54839fig1.jpg"/&gt; 图1:详细的手术过程的计划。 （一）腹股沟区解剖。 （B）收获上腹浅静脉移植。 （三）配售受体动物的动脉树桩之间的静脉移植。 （D）吻合针顺序。静脉移植后施工（E）站点。静脉移植干预的再灌注后（F）站点。黑色箭头标记胃脘浅静脉。的比例尺= 5毫米。 <a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/54839/54839fig1large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <img alt="图2" src="/files/ftp_upload/54839/54839fig2.jpg" /> 图2:动物模型的表征。静脉插入的（A）双功能超声嫁接IMMED<html> iately移植（0D）和28天术后（28D）之后。从0天及手术后的28日内移植到吕克负鼠吕克阳性的移植物（B）BLI。 21天7，14，后（C）的代表性的接枝截面收获，和28以及用Masson三色染色。 （上图:在比例尺= 400微米;下图:在比例尺= 100微米）（D）的百分比管腔闭塞通过从前者管腔除以新内腔计算。间差异通过方差分析（ANOVA）和Bonferroni的post-hoc检验的单向分析评估。 P &lt;0.01。误差棒为标准偏差（SD）。 <a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/54839/54839fig2large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> _upload / 54839 / 54839Sup1.jpg"/&gt; 补充文件1:手术过程的示意图。沿着LINEA inguinalis的垂直切口捐助大鼠进行，露出腹壁下静脉。血流停止用两个微钳，和静脉段长0.5-1.0厘米收获。在收件人大鼠，中位数股骨切口进行，暴露股静脉，动脉，神经。夹紧股动脉之后，血管段被除去，并通过将收获的静脉移植替代。 <a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/54839/Vein_interpostition_3.pptx" target="_blank">请点击此处查看补充文件1（或右键点击下载）。</a>

Watch this Scientific Journal Video about Vein Interposition Model: A Suitable Model to Study Bypass Graft Patency at JoVE.com

Vein Interposition Model: A Suitable Model to Study Bypass Graft Patency

This video demonstrates a model to study the development of myointimal hyperplasia after venous interposition surgery in rats.

静脉介入型号:研究旁路移植通畅一个合适的模型

Bypass grafting is an established treatment method for coronary artery disease. Graft patency continues to be the Achilles heel of saphenous vein grafts. ...

vein-interposition-model-suitable-model-to-study-bypass-graft

Nanyang Technological University

Research

JoVE Journal

Medicine

8.7K 次观看.  University Heart Center Hamburg. This video demonstrates a model to study the development of myointimal hyperplasia after venous interposition surgery in rats. 

视频: Vein Interposition Model: A Suitable Model to Study Bypass Graft Patency

Training a Sophisticated Microsurgical Technique: Interposition of External Jugular Vein Graft in the Common Carotid Artery in Rats

Neointimal hyperplasia is one the primary causes of stenosis in arterialized veins that are of great importance in arterial coronary bypass surgery, in peripheral arterial bypass surgery as well as in arteriovenous fistulas.1-5 The experimental procedure of vein graft interposition in the common carotid artery by using the cuff-technique has been applied in several research projects to examine the aetiology of neointimal hyperplasia and therapeutic options to address it. 6-8 The cuff prevents vessel anastomotic remodeling and induces turbulence within the graft and thereby the development of neointimal hyperplasia.
Using the superior caval vein graft is an established small-animal model for venous arterialization experiment.9-11 This current protocol refers to an established jugular vein graft interposition technique first described by Zou et al., 9 as well as others.12-14 Nevertheless, these cited small animal protocols are complicated.
To simplify the procedure and to minimize the number of experimental animals needed, a detailed operation protocol by video training is presented. This video should help the novice surgeon to learn both the cuff-technique and the vein graft interposition. Hereby, the right external jugular vein was grafted in cuff-technique in the common carotid artery of 21 female Sprague Dawley rats categorized in three equal groups that were sacrificed on day 21, 42 and 84, respectively. Notably, no donor animals were needed, because auto-transplantations were performed. The survival rate was 100 % at the time point of sacrifice. In addition, the graft patency rate was 60 % for the first 10 operated animals and 82 % for the remaining 11 animals. The blood flow at the time of sacrifice was 8&plusmn;3 ml/min. In conclusion, this surgical protocol considerably simplifies, optimizes and standardizes this complicated procedure. It gives novice surgeons easy, step-by-step instruction, explaining possible pitfalls, thereby helping them to gain expertise fast and avoid useless sacrifice of experimental animals.

Neointimal hyperplasia is the primary cause of stenosis in arterialized veins. We propose a new protocol whereby the right external jugular vein is grafted using the cuff technique in the common carotid artery of Sprague Dawley rats. The survival rate was 100 % at the time point of sacrifice.

培养一个精密的显微外科技术在大鼠颈总动脉，颈外静脉移植:介入

Neointimal hyperplasia is one the primary causes of stenosis in arterialized veins that are of great importance in arterial coronary bypass surgery, in ...

科研

医学

Adult Mouse Venous Hypertension Model: Common Carotid Artery to External Jugular Vein Anastomosis.

The understanding of the pathophysiology of brain arteriovenous malformations and arteriovenous fistulas has improved thanks to animal models. A rat model creating an artificial fistula between the common carotid artery (CCA) and the external jugular vein (EJV) has been widely described and proved technically feasible. This construct provokes a consistent cerebral venous hypertension (CVH), and therefore has helped studying the contribution of venous hypertension to formation, clinical symptoms, and prognosis of brain AVMs and dural AVFs. Equivalent mice models have been only scarcely described and have shown trouble with stenosis of the fistula. An established murine model would allow the study of not only pathophysiology but also potential genetic therapies for these cerebrovascular diseases.
We present a model of arteriovenous fistula that produces a durable intracranial venous hypertension in the mouse. Microsurgical anastomosis of the murine CCA and EJV can be difficult due to diminutive anatomy and frequently result in a non-patent fistula. In this step-by-step protocol we address all the important challenges encountered during this procedure. Avoiding excessive retraction of the vein during the exposure, using 11-0 sutures instead of 10-0, and making a carefully planned end-to-side anastomosis are some of the critical steps. Although this method requires advanced microsurgical skills and a longer learning curve that the equivalent in the rat, it can be consistently developed.
This novel model has been designed to integrate transgenic mouse techniques with a previously well-established experimental system that has proved useful to study brain AVMs and dural AVFs. By opening the possibility of using transgenic mice, a broader spectrum of valid models can be achieved and genetic treatments can also be tested. The experimental construct could also be further adapted to the study of other cerebrovascular diseases related with venous hypertension such as migraine, transient global amnesia, transient monocular blindness, etc.

We describe a method for creating a reliable model of cerebral venous hypertension in the adult mouse. This model has been widely described and tested in the rat. This new counterpart in the mice opens the possibility of using genetic modified animals and thereby broadens the applications of the model.

成年小鼠静脉高压型号:颈总动脉，以颈外静脉吻合。

The understanding of the pathophysiology of brain arteriovenous malformations and arteriovenous fistulas has improved thanks to animal models. A rat model ...

A Human Ex Vivo Atherosclerotic Plaque Model to Study Lesion Biology

Atherosclerosis is a chronic inflammatory disease of the vasculature. There are various methods to study the inflammatory compound in atherosclerotic lesions. Mouse models are an important tool to investigate inflammatory processes in atherogenesis, but these models suffer from the phenotypic and functional differences between the murine and human immune system. In vitro cell experiments are used to specifically evaluate cell type-dependent changes caused by a substance of interest, but culture-dependent variations and the inability to analyze the influence of specific molecules in the context of the inflammatory compound in atherosclerotic lesions limit the impact of the results. In addition, measuring levels of a molecule of interest in human blood helps to further investigate its clinical relevance, but this represents systemic and not local inflammation. Therefore, we here describe a plaque culture model to study human atherosclerotic lesion biology ex vivo. In short, fresh plaques are obtained from patients undergoing endarterectomy or coronary artery bypass grafting and stored in RPMI medium on ice until usage. The specimens are cut into small pieces followed by random distribution into a 48-well plate, containing RPMI medium in addition to a substance of interest such as cytokines or chemokines alone or in combination for defined periods of time. After incubation, the plaque pieces can be shock frozen for mRNA isolation, embedded in Paraffin or OCT for immunohistochemistry staining or smashed and lysed for western blotting. Furthermore, cells may be isolated from the plaque for flow cytometry analysis. In addition, supernatants can be collected for protein measurement by ELISA. In conclusion, the presented ex vivo model opens the possibility to further study inflammatory lesional biology, which may result in identification of novel disease mechanisms and therapeutic targets.

A Human Ex Vivo Atherosclerotic Plaque Model to Study Lesion Biology

Atherosclerosis is a chronic inflammatory process. This manuscript illustrates an easy to use ex vivo model to investigate fresh carotid or coronary artery plaques. The ex vivo model allows for the investigation of potential substances on the inflammatory milieu in human atherosclerotic lesions and results can be analyzed by various methods.

一个人力<em&gt;前体内</em&gt;动脉粥样硬化斑块模型研究病变的生物学

Atherosclerosis is a chronic inflammatory disease of the vasculature. There are various methods to study the inflammatory compound in atherosclerotic ...

Implantation of Inferior Vena Cava Interposition Graft in Mouse Model

Biodegradable scaffolds seeded with bone marrow mononuclear cells (BMCs) are often used for reconstructive surgery to treat congenital cardiac anomalies. The long-term clinical results showed excellent patency rates, however, with significant incidence of stenosis. To investigate the cellular and molecular mechanisms of vascular neotissue formation and prevent stenosis development in tissue engineered vascular grafts (TEVGs), we developed a mouse model of the graft with approximately 1 mm internal diameter. First, the TEVGs were assembled from biodegradable tubular scaffolds fabricated from a polyglycolic acid nonwoven felt mesh coated with &#949;-caprolactone and L-lactide copolymer. The scaffolds were then placed in a lyophilizer, vacuumed for 24 hr, and stored in a desiccator until cell seeding. Second, bone marrow was collected from donor mice and mononuclear cells were isolated by density gradient centrifugation. Third, approximately one million cells were seeded on a scaffold and incubated O/N. Finally, the seeded scaffolds were then implanted as infrarenal vena cava interposition grafts in C57BL/6 mice. The implanted grafts demonstrated excellent patency (&#62;90%) without evidence of thromboembolic complications or aneurysmal formation. This murine model will aid us in understanding and quantifying the cellular and molecular mechanisms of neotissue formation in the TEVG.

To improve our knowledge of cellular and molecular neotissue formation, a murine model of the TEVG was recently developed. The grafts were implanted as infrarenal vena cava interposition grafts in C57BL/6 mice. This model achieves similar results to those achieved in our clinical investigation, but over a far shortened time-course.

下腔静脉间置移植物在小鼠模型中植入

Biodegradable scaffolds seeded with bone marrow mononuclear cells (BMCs) are often used for reconstructive surgery to treat congenital cardiac anomalies. ...

Cardiopulmonary Bypass in a Mouse Model: A Novel Approach

As prolonged cardiopulmonary bypass becomes more essential during cardiac interventions, an increasing clinical demand arises for procedure optimization and for minimizing organ damage resulting from prolonged extracorporal circulation. The goal of this paper was to demonstrate a fully functional and clinically relevant model of cardiopulmonary bypass in a mouse. We report on the device design, perfusion circuit optimization, and microsurgical techniques. This model is an acute model, which is not compatible with survival due to the need for multiple blood drawings. Because of the range of tools available for mice (e.g., markers, knockouts, etc.), this model will facilitate investigation into the molecular mechanisms of organ damage and the effect of cardiopulmonary bypass in relation to other comorbidities.

This paper describes how to perform cardiopulmonary bypass in mice. This novel model will facilitate the investigation of the molecular mechanisms involved in organ damage.

小鼠体外循环模型: 一种新方法

As prolonged cardiopulmonary bypass becomes more essential during cardiac interventions, an increasing clinical demand arises for procedure optimization ...

Surgical Swine Model of Chronic Cardiac Ischemia Treated by Off-Pump Coronary Artery Bypass Graft Surgery

Chronic cardiac ischemia that impairs cardiac function, but does not result in infarct, is termed hibernating myocardium (HM). A large clinical subset of coronary artery disease (CAD) patients have HM, which in addition to causing impaired function, puts them at higher risk for arrhythmia and future cardiac events. The standard treatment for this condition is revascularization, but this has been shown to be an imperfect therapy. The majority of pre-clinical cardiac research focuses on infarct models of cardiac ischemia, leaving this subset of chronic ischemia patients largely underserved. To address this gap in research, we have developed a well-characterized and highly reproducible model of hibernating myocardium in swine, as swine are ideal translational models for human heart disease. In addition to creating this unique disease model, we have optimized a clinically relevant treatment model of coronary artery bypass surgery in swine. This allows us to accurately study the effects of bypass surgery on heart disease, as well as investigate additional or alternate therapies. This model surgically induces single vessel stenosis by implanting a constrictor on the left anterior descending (LAD) artery in a young pig. As the pig grows, the constrictor creates a gradual stenosis, resulting in chronic ischemia with impaired regional function, but preserving tissue viability. Following the establishment of the hibernating myocardium phenotype, we perform off-pump coronary artery bypass graft surgery to revascularize the ischemic region, mimicking the gold-standard treatment for patients in the clinic.

This protocol presents a surgical large animal model of chronic, single vessel ischemia that results in regional abnormalities but does not create infarct, known as hibernating myocardium. Following establishment of chronic ischemia, animals are treated with off-pump LIMA-LAD coronary artery bypass graft surgery to revascularize the ischemic tissue.

非体外循环冠状动脉旁路移植术治疗慢性心肌缺血的外科猪模型

Chronic cardiac ischemia that impairs cardiac function, but does not result in infarct, is termed hibernating myocardium (HM). A large clinical subset of ...

A Rabbit Model of Durable Transgene Expression in Jugular Vein to Common Carotid Artery Interposition Grafts

Vein graft bypass surgery is a common treatment for occlusive arterial disease; however, long-term success is limited by graft failure due to thrombosis, intimal hyperplasia, and atherosclerosis. The goal of this article is to demonstrate a method for placing bilateral venous interposition grafts in a rabbit, then transducing the grafts with a gene transfer vector that achieves durable transgene expression. The method allows the investigation of the biological roles of genes and their protein products in normal vein graft homeostasis. It also allows the testing of transgenes for the activities that could prevent vein graft failure, e.g., whether the expression of a transgene prevents the neointimal growth, reduces the vascular inflammation, or reduces atherosclerosis in rabbits fed with a high-fat diet. During an initial survival surgery, the segments of right and left external jugular vein are excised and placed bilaterally as reversed end-to-side common carotid artery interposition grafts. During a second survival surgery, performed 28 days later, each of the grafts is isolated from the circulation with vascular clips and the lumens are filled (via an arteriotomy) with a solution containing a helper-dependent adenoviral (HDAd) vector. After a 20-min incubation, the vector solution is aspirated, the arteriotomy is repaired, and flow is restored. The veins are harvested at time points dictated by individual experimental protocols. The 28-day delay between the graft placement and the transduction is necessary to ensure the adaptation of the vein graft to the arterial circulation. This adaptation avoids rapid loss of transgene expression that occurs in vein grafts transduced before or immediately after grafting. The method is unique in its ability to achieve durable, stable transgene expression in grafted veins. Compared to other large animal vein graft models, rabbits have advantages of low cost and easy handling. Compared to rodent vein graft models, rabbits have larger and easier-to-manipulate blood vessels that provide abundant tissue for analysis.

This method describes the placement of interposition vein grafts in rabbits, the transduction of the grafts, and the achievement of durable transgene expression. This allows the investigation of physiological and pathological roles of transgenes and their protein products in grafted veins, and testing of gene therapies for vein graft disease.

颈动脉介入移植中颈静脉持久性转基因表达的家兔模型

Vein graft bypass surgery is a common treatment for occlusive arterial disease; however, long-term success is limited by graft failure due to thrombosis, ...

Isometric Contractility Measurement of the Mouse Mesenteric Artery Using Wire Myography

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and drugs. It is widely used in many studies on the physiological functions of vascular smooth muscle, the pathogenesis of vascular diseases such as hypertension, and the development of smooth muscle relaxant drugs. The mouse is a widely used model animal with a large pool of disease models and genetically modified strains. We introduced this method to measure the isometric contraction of mouse mesenteric artery in detail. A 1.4-mm segment of mouse mesenteric resistance artery was isolated and mounted on a myograph chamber by passing two steel wires through its lumen. After equilibration and normalization steps, the vessel segment was potentiated by a high-K+ solution twice prior to the contraction assay. As an example of the application of this method in drug development, we measured the relaxant effect of a novel natural substance, neoliensinine, isolated from a Chinese herb, embryos of the lotus seed (Nelumbo nucifera Gaertn.) on mouse mesenteric arteries. The vessel segments mounted on the myograph chamber were stimulated with a high-K+ solution. When the force tension reached a stable sustained phase, cumulative doses of neoliensinine were added to the chamber. We found that neoliensinine had a dose-dependent relaxant effect on smooth muscle contraction, thus suggesting that it bears potential activity against hypertension. In addition, as the vessel segment can survive at least 4 hours after mounting and maintain contractility induced by the high-K+ solution for many times, we suggest that the wire myograph system may be used for the time-consuming process of drug screening.

The wire myograph technique is used to investigate vascular smooth muscle functions and screen new drugs. We report a detailed protocol for measuring the isometric contractility of the mouse mesenteric artery and for screening new relaxants of vascular smooth muscle.

用线 Myography 测定小鼠肠系膜动脉的等轴收缩力

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and ...

A Cryoinjury Model to Study Myocardial Infarction in the Mouse

The use of animal models is essential for developing new therapeutic strategies for acute coronary syndrome and its complications. In this article, we demonstrate a murine cryoinjury infarct model that generates precise infarct sizes with high reproducibility and replicability. In brief, after intubation and sternotomy of the animal, the heart is lifted from the thorax. The probe of a handheld liquid nitrogen delivery system is applied onto the myocardial wall to induce cryoinjury. Impaired ventricular function and electrical conduction can be monitored with echocardiography or optical mapping. Transmural myocardial remodeling of the infarcted area is characterized by collagen deposition and loss of cardiomyocytes. Compared to other models (e.g., LAD-ligation), this model utilizes a handheld liquid nitrogen delivery system to generate more uniform infarct sizes.

This article demonstrates a model to study cardiac remodeling after myocardial cryoinjury in mice.

研究小鼠心肌梗死的低温损伤模型

The use of animal models is essential for developing new therapeutic strategies for acute coronary syndrome and its complications. In this article, we ...

A Rabbit Venous Interposition Model Mimicking Revascularization Surgery using Vein Grafts to Assess Intimal Hyperplasia under Arterial Blood Pressure

Although vein grafts have been commonly used as autologous grafts in revascularization surgeries for ischemic diseases, the long-term patency remains poor because of the acceleration of intimal hyperplasia due to the exposure to arterial blood pressure. The present protocol is designed for the establishment of experimental venous intimal hyperplasia by interposing rabbit jugular veins to the ipsilateral carotid arteries. The protocol does not require surgical procedures deep in the body trunk and the extent of the incision is limited, which is less invasive for the animals, allowing long-term observation after implantation. This simple procedure enables researchers to investigate strategies to attenuate the progression of intimal hyperplasia of the implanted vein grafts. Using this protocol, we reported the effects transduction of microRNA-145 (miR-145), which is known to control the phenotype of vascular smooth muscle cells (VSMCs) from the proliferative to the contractile state, into harvested vein grafts. We confirmed the attenuation of intimal hyperplasia of vein grafts by transducing miR-145 before implantation surgery through the phenotype change of the VSMCs. Here we report a less invasive experimental platform to investigate the strategies that can be used to attenuate intimal hyperplasia of vein grafts in revascularization surgeries.

The present protocol aims to experimentally create venous intimal hyperplasia by subjecting veins to arterial blood pressure for developing strategies to attenuate venous intimal hyperplasia following revascularization surgery using vein grafts.

兔子静脉介词模型模仿再血管化手术使用静脉移植评估动脉血压下的影响增生

Although vein grafts have been commonly used as autologous grafts in revascularization surgeries for ischemic diseases, the long-term patency remains poor ...