Name: implantation of human-sized coronary stents into rat abdominal aorta using a trans-femoral access
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Description: Watch this Scientific Journal Video about Implantation of Human-Sized Coronary Stents into Rat Abdominal Aorta Using a Trans-Femoral Access at JoVE.com

我们要感谢安吉拉·弗伦德夫人在嵌入和幻灯片生产方面提供的宝贵技术援助。我们还要感谢实验室动物科学和实验外科研究所的塔杜斯·斯托平斯基先生在兽医工作中的有见地的帮助。

这些实验是根据德国动物福利法（TSchG）和2010/63/欧盟关于保护用于科学目的的动物的指令进行的。政府动物护理和使用委员会批准了这项研究的正式批准（第87-51.04.A065号议定书:兰德萨姆特·法尔·纳图尔，乌姆韦尔特·德·弗布劳彻舒茨·诺德海因-韦斯特法伦，德国雷克林豪森）。研究协议符合《实验室动物护理和使用指南》。术后疼痛治疗基于德国实验室动物科学协会 （GV-SOLAS） 的建议以及主动兽医疼痛治疗。
1. 基本技术和常见程序
<ol><li>使用同源猿-/- 斯普拉格-道利大鼠。使用标准方法17识别每种动物的基因型。</li>
	<li>将动物保持在相同的条件下（21 °C ± 2 °C，60%± 5% 的湿度，以及 12 小时的光/暗循环），并确保免费获得水和食物。</li>
	<li>在清洁但非星号条件下执行所有程序。</li>
	<li>一旦大鼠麻醉，在手术显微镜下执行所有手术，倍数为16倍。</li>
	<li>使用棉签进行压缩溶血。用乳化铃声溶液浸泡的纱布（5 厘米 x 5 厘米）有助于保持腹股沟湿润。</li>
	<li>遵守废物处理条例，处理废旧材料。</li>
</ol>2. 手术前的准备
<ol><li>在开始操作之前准备兽药。除非另有说明，否则请在室温下保留所有解决方案。</li>
	<li>
手术前30分钟，皮下施用 0.03-0.05毫克/千克丁丙诺啡。
</li>
	<li>麻醉大鼠，注射100毫克/千克体重（BW）和8毫克/千克BW西拉辛。</li>
	<li>使用称重秤评估大鼠的体重。</li>
	<li>将鼠放在加热垫上，用医用胶带固定上下肢。将大鼠的左后肢完全伸展，并尽可能与脊椎保持一致，从而在股动脉和主动脉之间形成一条直线。这将促进气球安装支架通过主动脉分叉前进。</li>
	<li>保持麻醉，吸入1.5伏的异氟烷在97.5%的氧气中，流量为2L/min。 注意:允许大鼠自发呼吸，无需管网。</li>
	<li>应用眼药膏，防止无意识时眼睛损伤。</li>
	<li>从大鼠腹股沟和下腹部区域剃毛，用波维酮碘溶液对相应的皮肤进行消毒。</li>
	<li>在开始手术之前，通过捏紧尾尖和分形组织来验证麻醉的深度。</li>
</ol>3. 手术
<ol><li>在左腹股沟中切开约 0.5+u20121 厘米，以打开皮肤和底层筋膜。</li>
	<li>在深部进行模糊解剖和探查，直到可以识别脉动的左股动脉。</li>
	<li>使用非常精细的钳子，通过轻轻取出周围的结缔组织来准备股骨动脉。小心伤害股骨神经和股骨静脉，这是动脉中间。</li>
	<li>准备大约1厘米的股骨动脉。小心地将钳尖放在容器下，轻轻抬起。</li>
	<li>线条4-0丝缝合线下动脉的解剖和接近部分，并形成吊索。夹住手术夹的分支之间的两根螺纹吊索的末端。使用手术夹控制动脉。轻轻伸展和抬起吊索，以暂时中断血液流动。 注意:快速工作，以避免可能导致组织损伤的长时间旅游。</li>
	<li>使用锋利的微剪刀，在股动脉中间进行动脉切除术。</li>
	<li>通过动脉切除术引入导线。当到达近线吊索时，通过移动手术夹释放螺纹的张力，并将导线进一步向腹部主动脉推进。 注意:使用切割机切割导线以便于操作。</li>
	<li>将导线的近端放置在隔膜和肾动脉之间。 注意:将导线推进太远会承受主动脉或心脏损伤的风险。我们建议打开腹部，以确保引导线和支架的充分定位，至少对于前几只动物。</li>
	<li>在导引线上引入一个压接式和气球式冠状动脉支架，尺寸为 2.25 mm x 8 mm（最大 2.5 mm x 8 mm），并将其推进到腹动脉。</li>
	<li>将支架放在主动脉分叉上方，但放在肾动脉下方。使用充气注射器系统将气球导管充气至 12 atm 15s，从而部署支架。</li>
	<li>根据制造商对使用支架的建议，放气气球导管并保持负压。</li>
	<li>在将支架留在原位时，慢慢取出放气导管。</li>
	<li>在取出导管之前，用手术夹在切口上方的螺纹环上制造张力，再次中断血液流动。然后取出气球导管，直接将容器靠近。</li>
	<li>将近部和解剖线环系成石灰，以盖住股动脉，并确认动脉切除术的充分血吸虫。辅助动脉将确保进一步灌注到肢体。</li>
	<li>使用 10-0 非可恢复缝合，关闭动脉的肌肉以及皮肤切口。</li>
</ol>4. 支架植入后的动物护理
<ol><li>手术后，让老鼠在特殊的重症监护室笼子里恢复60分钟，笼子里有温暖的空气（30\u201235°C）和氧气供应。</li>
	<li>仔细观察动物，直到完全康复。之后，将老鼠移入普通笼子。提供获得水和食物的辅助通道。</li>
	<li>根据临床评估，每6-12小时服用一次术后镇痛，服用0.03-0.05毫克/千克丁丙诺啡（s.c.，500μl NaCl），共72小时。</li>
	<li>将食物与氯皮多格雷（15毫克/千克）混合，以避免植入支架的血栓形成。</li>
	<li>为了改善高血糖条件和斑块形成，在出生后6\u20128周开始西方饮食喂养，一直持续到安乐死。如果需要，一群喂养正常老鼠的动物可以作为控制。</li>
</ol>5. 组织收集和处理
<ol><li>在指定时间点开始组织去除之前，根据 IACUC 指南对动物实施安乐死。在观察期结束时收获被强化的主动脉进行病理分析。</li>
	<li>通过中线切口打开腹部，取出主动脉的固定部分以及主动脉的相邻非固定部分，每个部分尺寸为 0.5 厘米。</li>
	<li>将组织放入 4% 缓冲形式素的溶液中，以进行 24 小时的固定。</li>
	<li>将被强化的动脉组织嵌入塑料中，并按照标准协议18、19进行组织学和免疫化学染色。</li>
</ol>6. 组织学分析
<ol><li>通过与计算机相连的显微镜，使用适当的图像分析软件，对紧固主动脉的近、中、离部分的顺序部分进行成形分析。</li>
	<li>使用图形绘图板跟踪外部弹性拉米纳（EEL，在冒险和介质之间）、内部弹性拉米纳（IEL、介质和新因蒂玛之间）和流明的轮廓。从这些值中，使用软件计算 EEL 区域、IEL 区域和流明区域。</li>
	<li>计算支架内复发 （ISR） 的横截面面积百分比: <img alt="Equation 1" src="/files/ftp_upload/61442/61442equ01.jpg">
</li>
	<li>计算新近面积（Ai）: <img alt="Equation 2" src="/files/ftp_upload/61442/61442equ02.jpg">
</li>
	<li>测量每个支架支柱上的新意厚度 （NIT），作为支柱和流明之间的距离。测量支架支柱之间的NIT，作为 IEL 和流明之间的距离。 注:或者，将NIT计算为 <img alt="Equation 3" src="/files/ftp_upload/61442/61442equ03.jpg"> 其中PL和PIEL是流明和内部弹性拉米纳周长，分别为20。</li>
	<li>根据研究的要求进行额外的分析。</li>
</ol>

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M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Early+adaptations+in+collateral+and+microvascular+resistances+after+ligation+of+the+rat+femoral+artery.">Early adaptations in collateral and microvascular resistances after ligation of the rat femoral artery.</a> Journal of Applied Physiology. 79 (1), 73-82 (1985).</li><li>Nevzati, E., 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=Biodegradable+Magnesium+Stent+Treatment+of+Saccular+Aneurysms+in+a+Rat+Model+-+Introduction+of+the+Surgical+Technique.">Biodegradable Magnesium Stent Treatment of Saccular Aneurysms in a Rat Model - Introduction of the Surgical Technique.</a> Journal of Visualized Experiments. (128), e56359 (2017).</li><li>Aquarius, R., Smits, D., Gounis, M. J., Leenders, W. P. J., de Vries, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Flow+diverter+implantation+in+a+rat+model+of+sidewall+aneurysm:+a+feasibility+study.">Flow diverter implantation in a rat model of sidewall aneurysm: a feasibility study.</a> Journal of NeuroInterventional Surgery. 10 (1), 88-92 (2018).</li></ol>

作者没有什么可透露的。

本协议描述了将人类大小的冠状动脉支架植入大鼠腹 主动脉。几个技术点值得强调。首先，应避免支架大小和主动脉大小之间的不匹配。放置过小的支架会导致支架失调，而植入对主动脉来说太大的支架则会导致血管过度拉伸、撕裂和损伤。因此，我们建议使用直径在 2.0 到 2.5 毫米之间的支架，并将植入压力保持在建议范围内，而不要过度拉伸支架。最合适的植入压力通常由支架制造商...

皮下冠心病干预（PCI），结合冠状支架的部署，代表了冠状动脉疾病干预治疗的黄金标准1。然而，支架的长期成功可以限制在支架内复发（ISR）的发生，这是由支架2，3内新意组织过度增殖决定的。ISR 可能需要用冠状动脉旁路或重新 PCI 进行重新干预。为研究ISR提出了各种动物模型的建议，每个模型都有优点和缺点。最常用的猪冠状动脉和兔肠动脉模型的主要缺点，虽然在支架植入4、5后出现与人类明显相似的病变，但动物和住房成本高，这带来了后勤困难，特别是在长期研究中，以及处理和设备的限制。此外，猪和兔子细胞蛋白抗体的可用性有限。另一方面，Murine 模型具有吞吐量高、可重复性高、易于操作、外壳等主要优点，因此具有成本效益。此外，有较多的抗体可用。然而，虽然非丙蛋白电子缺乏症（apoE-/-）小鼠已被广泛用于动脉粥样硬化6，7，8的研究，他们不适合研究ISR，因为支架必须小型化植入小鼠，有可能改变支架的机械特性。此外，小鼠主动脉壁测量在幼鼠50μm和老老鼠9中85μm之间，支架必须使用低至2 atm的压力水平进行部署，这可能导致支架10的不良位。然而，大鼠允许植入市售的人类冠状动脉支架，并表现出类似于主动脉支架植入后较大的动物的血管愈合过程，首先由Langeveld等人11日报道。这项技术最初需要跨腹部通道，这就需要对主动脉进行物理收缩，以实现血液流动的暂时中断。为了避免潜在的相关船只伤害和炎症反应，该技术后来通过引入一个跨组织访问，这进一步导致动物12的存活率更高。
由于野生类型大鼠不发展动脉粥样硬化病变13，apoE-/-大鼠已产生使用核酶技术，如转录激活器一样的影响器核糖核酸（TALEN）14，聚类定期间歇短巴林德罗米重复（CRISPR/Cas9）15，和锌手指（ZF）16。自 2011 年以来，ApoE-/-大鼠已上市。提供外来背景，apoE-/-大鼠允许更现实地评估人类大小的冠状动脉支架，特别是关于ISR。
在此，我们通过跨性别访问路线描述该方法，并使用市售的薄支柱钴-铬药物脱脂支架 （DES），但是，它也可用于研究其他支架类型，如裸金属支架 （BMS） 或可生物降解支架。

<table><tbody><tr><td>&lt;strong&gt;Diet&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>SNIFF High Fat diet + Clopidogrel (15 mg/kg)</td><td>SNIFF Spezialdi&amp;auml;ten GmbH, Soest</td><td>custom prepared</td><td>Western Diet</td></tr><tr><td></td><td></td><td></td><td></td></tr><tr><td>&lt;strong&gt;Drugs and Anesthetics&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>Buprenorphine</td><td>Essex Pharma</td><td>997.00.00</td><td></td></tr><tr><td>ISOFLO (Isoflurane Vapor) vaporiser</td><td>Eickemeyer</td><td>4802885</td><td></td></tr><tr><td>Isoflurane</td><td>Forene Abbott</td><td>B 506</td><td></td></tr><tr><td>Isotonic (0.9%) NaCl solution</td><td>DeltaSelect GmbH</td><td>PZN 00765145</td><td></td></tr><tr><td>Ringer&#039;s lactate solution</td><td>Baxter Deutschland GmbH</td><td>3775380</td><td></td></tr><tr><td>(S)-ketamine</td><td>CEVA Germany</td><td></td><td></td></tr><tr><td>Xylazine</td><td>Medistar Germany</td><td></td><td></td></tr><tr><td></td><td></td><td></td><td></td></tr><tr><td>&lt;strong&gt;Consumable supplies&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>10 mL syringes</td><td>BD Plastipak</td><td>4606108V</td><td></td></tr><tr><td>2 mL syringes</td><td>BD Plastipak</td><td>4606027V</td><td></td></tr><tr><td>6-0 prolene suture</td><td>ETHICON</td><td>N-2719K</td><td></td></tr><tr><td>4-0 silk suture</td><td>Seraflex</td><td>IC 158000</td><td></td></tr><tr><td>Bepanthen Eye and Nose Ointment</td><td>Bayer Vital GmbH</td><td>6029009.00.00</td><td></td></tr><tr><td>Cotton Gauze swabs</td><td>Fuhrmann GmbH</td><td>32014</td><td></td></tr><tr><td>Durapore silk tape</td><td>3M</td><td>1538-1</td><td></td></tr><tr><td>Poly-Alcohol Skin Desinfection Solution</td><td>Antiseptica GmbH</td><td>72PAH200</td><td></td></tr><tr><td>Sterican needle 18 G</td><td>B. Braun</td><td>304622</td><td></td></tr><tr><td>Sterican needle 27 3/4 G</td><td>B.Braun</td><td>4657705</td><td></td></tr><tr><td>Tissue Paper</td><td>commercially available</td><td></td><td></td></tr><tr><td></td><td></td><td></td><td></td></tr><tr><td>&lt;strong&gt;Surgical instruments&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>Graefe forceps curved x1</td><td>Fine Science Tools Inc.</td><td>11151-10</td><td></td></tr><tr><td>Graefe forceps straight</td><td>Fine Science Tools Inc.</td><td>11050-10</td><td></td></tr><tr><td>Needle holder Mathieu</td><td>Fine Science Tools Inc.</td><td>12010-14</td><td></td></tr><tr><td>Scissors</td><td>Fine Science Tools Inc.</td><td>14074-11</td><td></td></tr><tr><td>Semken forceps</td><td>Fine Science Tools Inc.</td><td>11008-13</td><td></td></tr><tr><td>Small surgical scissors curved</td><td>Fine Science Tools Inc.</td><td>14029-10</td><td></td></tr><tr><td>Small surgical scissors straight</td><td>Fine Science Tools Inc.</td><td>14028-10</td><td></td></tr><tr><td>Standard pattern forceps</td><td>Fine Science Tools Inc.</td><td>11000-12</td><td></td></tr><tr><td>Vannas spring scissors</td><td>Fine Science Tools Inc.</td><td>15000-08</td><td></td></tr><tr><td></td><td></td><td></td><td></td></tr><tr><td>&lt;strong&gt;Equipment&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>Dissecting microscope</td><td>Leica MZ9</td><td></td><td></td></tr><tr><td>Temperature controlled heating pad</td><td>Sygonix</td><td>26857617</td><td></td></tr><tr><td></td><td></td><td></td><td></td></tr><tr><td>&lt;strong&gt;Equipment for stent implantation&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>Drug-eluting stent Xience 2,25mm x 8mm</td><td>Abbott Vascular USA</td><td>1009544-18</td><td></td></tr><tr><td>Guide wire Fielder XT PTCA guide wire: 0.014&quot; x 300cm</td><td>ASAHI INTECC CO., LTD Japan</td><td>AGP140302</td><td></td></tr><tr><td>Inflation syringe system</td><td>Abbott 20/30 Priority Pack</td><td>1000186</td><td></td></tr><tr><td></td><td></td><td></td><td></td></tr><tr><td>&lt;strong&gt;Tissue processing and analysis&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>30% H&lt;sub&gt;2&lt;/sub&gt;O&lt;sub&gt;2&lt;/sub&gt;</td><td>Roth</td><td>9681</td><td>Histology</td></tr><tr><td>Ethanol</td><td>Roth</td><td>K928.1</td><td>Histology</td></tr><tr><td>Giemsas Azur-Eosin-Methylenblau</td><td>Merck</td><td>109204</td><td>Histology</td></tr><tr><td>Graphic Drawing Tablet</td><td>WACOM Europe GmbH</td><td>CTL-6100WLK-S</td><td></td></tr><tr><td>Roti Histofix, Formaldehyd 4% buffered</td><td>Roth</td><td>P087</td><td>Histology</td></tr><tr><td>Technovit 9100</td><td>Morphisto</td><td>12225.K1000</td><td>Histology</td></tr></tbody></table>

implantation of human-sized coronary stents into rat abdominal aorta using a trans-femoral access

皮下冠心病干预 （PCI） 结合冠状支架的部署，代表了冠状动脉疾病干预治疗的黄金标准。支架内恢复症 （ISR） 是由支架内新辛蒂莫组织过度增殖决定的，并限制支架的长期成功。各种动物模型被用来阐明支架内复发 （ISR） 背后的病理生理过程，其中猪冠状动脉和兔子胆道模型最常使用。Murine 模型具有吞吐量高、易于处理和外壳、可重复性和分子标记的广泛可用性等优点。非丙蛋白E缺乏（apoE-/-）小鼠模型已广泛用于研究心血管疾病。然而，支架必须小型化才能植入小鼠体内，这涉及到其机械和（潜在）生物特性的重大变化。使用猿类-/-大鼠可以克服这些缺点，因为apoE-/-大鼠允许评估人类大小的冠状动脉支架，同时提供一个异位表型。这使得它们成为在支架植入后研究 ISR 的极好可靠的模型。在这里，我们详细描述了将市售的人类冠状动脉支架植入具有apoE-/-背景的老鼠的腹部主动脉，使用跨股骨访问。

该协议描述了使用跨股骨通道（图1）植入大鼠腹部主动脉的支架。这种动物模型的第一个中心点是，它允许部署人类大小的冠状动脉支架。一个市售的压接和气球安装冠状动脉支架可以放置到大鼠的腹部主动脉。因此，还可以应用与人类相同的支架部署原则。使用大鼠的另一个优点是提供转基因菌株，如apoE-/- 大鼠，这些菌株在商业上是可用的。
<p class="jove_conten...

Watch this Scientific Journal Video about Implantation of Human-Sized Coronary Stents into Rat Abdominal Aorta Using a Trans-Femoral Access at JoVE.com

Implantation of Human-Sized Coronary Stents into Rat Abdominal Aorta Using a Trans-Femoral Access

该协议描述了人类冠状动脉支架植入大鼠的腹部主动脉与apoE-/- 背景使用跨股骨访问。与其他动物模型相比，穆林模型具有吞吐量高、可重复性高、易处理性和外壳性好等优点，分子标记物广泛可用。

使用跨女性访问将人类大小的冠状动脉支架植入大鼠腹主动脉

Percutaneous coronary intervention (PCI), combined with the deployment of a coronary stent, represents the gold standard in interventional treatment of ...

implantation-human-sized-coronary-stents-into-rat-abdominal-aorta

Research

JoVE Journal

Medicine

4.2K 次观看.  University Hospital RWTH Aachen. 该协议描述了人类冠状动脉支架植入大鼠的腹部主动脉与apoE-/- 背景使用跨股骨访问。与其他动物模型相比，穆林模型具有吞吐量高、可重复性高、易处理性和外壳性好等优点，分子标记物广泛可用。

视频: Implantation of Human-Sized Coronary Stents into Rat Abdominal Aorta Using a Trans-Femoral Access

Orthotopic Aortic Transplantation: A Rat Model to Study the Development of Chronic Vasculopathy

Research models of chronic rejection are essential to investigate pathobiological and pathophysiological processes during the development of transplant vasculopathy (TVP).
The commonly used animal model for cardiovascular chronic rejection studies is the heterotopic heart transplant model performed in laboratory rodents. This model is used widely in experiments since Ono and Lindsey (3) published their technique. To analyze the findings in the blood vessels, the heart has to be sectioned and all vessels have to be measured.
Another method to investigate chronic rejection in cardiovascular questionings is the aortic transplant model (1, 2). In the orthotopic aortic transplant model, the aorta can easily be histologically evaluated (2). The PVG-to-ACI model is especially useful for CAV studies, since acute vascular rejection is not a major confounding factor and Cyclosporin A (CsA) treatment does not prevent the development of CAV, similar to what we find in the clinical setting (4). A7-day period of CsA is required in this model to prevent acute rejection and to achieve long-term survival with the development of TVP.
This model can also be used to investigate acute cellular rejection and media necrosis in xenogeneic models (5).

This video demonstrates the orthotopic aortic transplant model as a simple model to study the development of transplant vasculopathy (TVP) in rats.

主动脉原位移植大鼠模型，研究慢性血管病变的发展

Research models of chronic rejection are essential to investigate pathobiological and pathophysiological processes during the development of transplant ...

科研

医学

Human Internal Mammary Artery (IMA) Transplantation and Stenting: A Human Model to Study the Development of In-Stent Restenosis

Preclinical in vivo research models to investigate pathobiological and pathophysiological processes in the development of intimal hyperplasia after vessel stenting are crucial for translational approaches1,2.
The commonly used animal models include mice, rats, rabbits, and pigs3-5. However, the translation of these models into clinical settings remains difficult, since those biological processes are already studied in animal vessels but never performed before in human research models6,7. In this video we demonstrate a new humanized model to overcome this translational gap. The shown procedure is reproducible, easy, and fast to perform and is suitable to study the development of intimal hyperplasia and the applicability of diverse stents.
This video shows how to perform the stent technique in human vessels followed by transplantation into immunodeficient rats, and identifies the origin of proliferating cells as human.

Human Internal Mammary Artery IMA Transplantation and Stenting: A Human Model to Study the Development of In-Stent Restenosis

This video shows a model to study the development of intimal hyperplasia after stent deployment using a human vessel (IMA) in an immunodeficient rat model.

人乳内动脉（IMA）移植及内支架置入术：人体模型支架内再狭窄的研究开发

Preclinical in vivo research models to investigate pathobiological and pathophysiological processes in the development of intimal hyperplasia after vessel ...

Implantation of Engineered Tissue in the Rat Heart

Rodent surgery is often an important component in assessing the utility of engineered tissues. A wide variety of surgical procedures can be performed in common laboratory rats or mice and these quite frequently serve as an intermediate step between bench-top experiments and large animal testing or human trials. Given that rodents provide an established, cost-effective, and physiologically-relevant model system in which to test novel combinations of scaffolding materials and cells, they are particularly well-suited for cardiovascular tissue engineering studies. Presently, we describe an open-heart surgical procedure to implant engineered tissue containing myogenic progenitor cells in the atrioventricular (AV) groove of a rat heart. These implants are intended to create an electrical conduit between the right atrium and right ventricle with the ultimate goal of providing an alternative treatment to conventional pacemaker implantation in pediatric patients with complete heart block[1]. The engineered tissue is implanted in the AV-groove by means of a thoracotomy. For our purposes, Lewis rats are anesthetized and invasively ventilated to maintain positive airway pressure during the sterile surgical procedure. The approach to the heart is performed by a right thoracotomy through an antero-lateral incision at the 5th intercostal space. The tissue construct is fixed in the AV groove using a single 7-0 Prolene suture and positioned between the right ventricle and atrium at the ventral portion of the heart. The epicardium is partially removed to allow direct contact between the recipient myocardial cells and those contained in the engineered tissue. Following implantation, the chest wall is closed in layers, any pneumothorax is evacuated, and the animal is extubated and treated with analgesic.

Here, we describe a cardiac surgical procedure to implant engineered tissue in the atrioventricular (AV)-groove of an adult Lewis rat.

大鼠心脏中植入的工程组织

Rodent surgery is often an important component in assessing the utility of engineered tissues. A wide variety of surgical procedures can be performed in ...

生物学

A Murine Model of Stent Implantation in the Carotid Artery for the Study of Restenosis

Despite the considerable progress made in the stent development in the last decades, cardiovascular diseases remain the main cause of death in western countries. Beside the benefits offered by the development of different drug-eluting stents, the coronary revascularization bears also the life-threatening risks of in-stent thrombosis and restenosis. Research on new therapeutic strategies is impaired by the lack of appropriate methods to study stent implantation and restenosis processes. Here, we describe a rapid and accessible procedure of stent implantation in mouse carotid artery, which offers the possibility to study in a convenient way the molecular mechanisms of vessel remodeling and the effects of different drug coatings.

A model of stent implantation in mouse carotid artery is described. Compared to other similar methods, this procedure is very rapid, simple and accessible, offering the possibility to study in a convenient way the vascular wall reaction to different drug-eluting stents and the molecular mechanisms of restenosis.

颈动脉支架植入再狭窄的研究的小鼠模型

Despite  the considerable progress made in the stent development in the last decades,  cardiovascular diseases remain the main cause of death in western ...

A New Murine Model of Endovascular Aortic Aneurysm Repair

Endovascular aneurysm exclusion is a validated technique to prevent aneurysm rupture. Long-term results highlight technique limitations and new aspects of Abdominal aortic aneurysm (AAA) pathophysiology. There is no abdominal aortic aneurysm endograft exclusion model cheap and reproducible, which would allow deep investigations of AAA before and after treatment. We hereby describe how to induce, and then to exclude with a covered coronary stentgraft an abdominal aortic aneurysm in a rat. The well known elastase induced AAA model was first reported in 19901 in a rat, then described in mice2. Elastin degradation leads to dilation of the aorta with inflammatory infiltration of the abdominal wall and intra luminal thrombus, matching with human AAA. Endovascular exclusion with small covered stentgraft is then performed, excluding any interactions between circulating blood and the aneurysm thrombus. Appropriate exclusion and stentgraft patency is confirmed before euthanasia by an angiography thought the left carotid artery. Partial control of elastase diffusion makes aneurysm shape different for each animal. It is difficult to create an aneurysm, which will allow an appropriate length of aorta below the aneurysm for an easy stentgraft introduction, and with adequate proximal and distal neck to prevent endoleaks. Lots of failure can result to stentgraft introduction which sometimes lead to aorta tear with pain and troubles to stitch it, and endothelial damage with post op aorta thrombosis. Giving aspirin to rats before stentgraft implantation decreases failure rate without major hemorrhage. Clamping time activates neutrophils, endothelium and platelets, and may interfere with biological analysis.

Histological and biochemical modifications after aneurysm endograft exclusion are still unclear. We describe a new model of endograft implantation on a murine aneurysm. Through thrombus analysis with and without persistant circulating blood stream interface, and new endovascular biomaterials evaluation, this model will help to better understand endovascular aneurysm exclusion pathobiology.

一种新的小鼠模型主动脉瘤腔内修复

Endovascular aneurysm exclusion is a validated technique to prevent aneurysm rupture. Long-term results highlight technique limitations and new aspects of ...

Imaging In-Stent Restenosis: An Inexpensive, Reliable, and Rapid Preclinical Model

Preclinical models of restenosis are essential to unravel the pathophysiological processes that lead to in-stent restenosis and to optimize existing and future drug-eluting stents.
A variety of antibodies and transgenic and knockout strains are available in rats. Consequently, a model for in-stent restenosis in the rat would be convenient for pathobiological and pathophysiological studies. 
In this video, we present the full procedure and pit-falls of a rat stent model suitable for high throughput stent research. We will show the surgical procedure of stent deployment, and the assessment of in-stent restenosis using the most elegant technique of OCT (Optical Coherence Tomography). This technique provides high accuracy in assessing plaque CSAs (cross section areas) and correlates well with histological sections, which require special and time consuming embedding and sectioning techniques. OCT imaging further allows longitudinal monitoring of the development of in-stent restenosis within the same animal compared to one-time snapshots using histology.

This video demonstrates how to use a preclinical inexpensive and reliable model to study pathobiological and pathophysiological processes of in-stent restenosis development. Longitudinal in vivo monitoring using OCT (Optical Coherence Tomography) and analysis of OCT images are also demonstrated.

支架内再狭窄的影像：一种廉价的，可靠，快速的临床前模型

Preclinical models of restenosis are essential to unravel the pathophysiological processes that lead to in-stent restenosis and to optimize existing and ...

Biodegradable Magnesium Stent Treatment of Saccular Aneurysms in a Rat Model - Introduction of the Surgical Technique

The steady progess in the armamentarium of techniques available for endovascular treatment of intracranial aneurysms requires affordable and reproducable experimental animal models to test novel embolization materials such as stents and flow diverters. The aim of the present project was to design a safe, fast, and standardized surgical technique for stent assisted embolization of saccular aneurysms in a rat animal model.
Saccular aneurysms were created from an arterial graft from the descending aorta.The aneurysms were microsurgically transplanted through end-to-side anastomosis to the infrarenal abdominal aorta of a syngenic male Wistar rat weighing &#62;500 g. Following aneurysm anastomosis, aneurysm embolization was performed using balloon expandable magnesium stents (2.5 mm x 6 mm). The stent system was retrograde introduced from the lower abdominal aorta using a modified Seldinger technique.
Following a pilot series of 6 animals, a total of 67 rats were operated according to established standard operating procedures. Mean surgery time, mean anastomosis time, and mean suturing time of the artery puncture site were 167 &#177; 22 min, 26 &#177; 6 min and 11 &#177; 5 min, respectively. The mortality rate was 6% (n=4). The morbidity rate was 7.5% (n=5), and in-stent thrombosis was found in 4 cases (n=2 early, n=2 late in stent thrombosis).
The results demonstrate the feasibility of standardized stent occlusion of saccular sidewall aneurysms in rats - with low rates of morbidity and mortality. This stent embolization procedure combines the opportunity to study novel concepts of stent or flow diverter based devices as well as the molecular aspects of healing.

Reproducable experimental animal models are needed for the testing of novel embolization materials, which have been designed to treat endovascular occlusion of intracranial aneurysms (IA). The present study aims to develop a safe and standardized surgical technique for stent assisted embolization of saccular aneurysms in a rat animal model.

生物可降解镁支架治疗大鼠囊动脉瘤的实验研究--手术技术介绍

The steady progess in the armamentarium of techniques available for endovascular treatment of intracranial aneurysms requires affordable and reproducable ...

神经科学

Novel Percutaneous Approach for Deployment of 3D Printed Coronary Stenosis Implants in Swine Models of Ischemic Heart Disease

Minimally invasive methods for creating models of focal coronary narrowing in large animals are challenging. Rapid prototyping using three-dimensionally (3D) printed coronary implants can be employed to percutaneously create a focal coronary stenosis. However, reliable delivery of the implants can be difficult without the use of ancillary equipment. We describe the use of a mother-and-child coronary guide catheter for stabilization of the implant and for effective delivery of the 3D printed implant to any desired location along the length of the coronary vessel. The focal coronary narrowing was confirmed under coronary cineangiography and the functional significance of the coronary stenosis was assessed using gadolinium-enhanced first-pass cardiac perfusion MRI. We showed that reliable delivery of 3D printed coronary implants in swine models (n = 11) of ischemic heart disease can be achieved through repurposing mother-and-child coronary guide catheters. Our technique simplifies the percutaneous delivery of coronary implants to create closed-chest swine models of focal coronary artery stenosis and can be performed expeditiously, with a low procedural failure rate.

We describe a novel, cost-effective, and efficient technique for percutaneous delivery of three-dimensionally printed coronary implants to create closed-chest swine models of ischemic heart disease. The implants were fixed in place using a mother-and-child extension catheter with high success rate.

在缺血性心脏病的猪模型中部署3D打印冠状动脉狭窄植入物的新颖的皮下方法

Minimally invasive methods for creating models of focal coronary narrowing in large animals are challenging. Rapid prototyping using three-dimensionally ...

Using a Cell-Tracer Injection to Investigate the Origin of Neointima-Forming Cells in a Rat Saccular Side Wall Model

Microsurgical clipping creates a subsequent barrier of blood flow into intracranial aneurysms, whereas endovascular treatment relies on neointima and thrombus formation. The source of endothelial cells covering the endoluminal layer of the neointima remains unclear. Therefore, the aim of the present study was to investigate the origin of neointima-forming cells after cell-tracer injection in the already well-established Helsinki rat microsurgical sidewall aneurysm model.
Sidewall aneurysms were created by suturing decellularized or vital arterial pouches end-to-side to the aorta in male Lewis rats. Before arteriotomy with aneurysm suture, a cell-tracer injection containing CM-Dil dye was performed into the clamped aorta to label endothelial cells in the adjacent vessel and track their proliferation during follow-up (FU). Treatment followed by coiling (n = 16) or stenting (n = 15). At FU (7 days or 21 days), all rats underwent fluorescence angiography, followed by aneurysm harvesting and macroscopic and histological evaluation with immunohistological cell counts for specific regions of interest. 
None of the 31 aneurysms had ruptured upon follow-up. Four animals died prematurely. Macroscopically residual perfusion was observed in 75.0% coiled and 7.0% of stented rats. The amount of cell-tracer-positive cells was significantly elevated in decellularized stented compared to coiled aneurysms with respect to thrombus on day 7 (p = 0.01) and neointima on day 21 (p = 0.04). No significant differences were found in thrombus or neointima in vital aneurysms. 
These findings confirm worse healing patterns in coiled compared to stented aneurysms. Neointima formation seems particularly dependent on the parent artery in decellularized aneurysms, whereas it is supported by the recruitment from aneurysm wall cells in vital cell-rich walls. In terms of translation, stent treatment might be more appropriate for highly degenerated aneurysms, whereas coiling alone might be adequate for aneurysms with mostly healthy vessel walls.

We performed a one-point, lipophilic cell-tracer injection to track endothelial cells, followed by an arteriotomy and suturing of sidewall aneurysms on the abdominal rat aorta. Neointima formation seemed dependent on the parent artery in decellularized aneurysms and was promoted by the recruitment from aneurysm wall cells in vital cell-rich walls.

使用细胞示踪剂注射研究大鼠囊侧壁模型中新内膜形成细胞的起源

Microsurgical clipping creates a subsequent barrier of blood flow into intracranial aneurysms, whereas endovascular treatment relies on neointima and ...

Implantation of Electrospun Vascular Grafts with Optimized Structure in a Rat Model

Here, we present a protocol to fabricate macroporous PCL vascular graft and describe an evaluation protocol by using a rat model of abdominal aorta replacement. The electrospun vascular grafts often possess relatively small pores, which limit cell infiltration into the grafts and hinder the regeneration and remodeling of the neo-arteries. In this study, PCL vascular grafts with thicker fibers (5 - 6 &#181;m) and larger pores (~30 &#181;m) were fabricated by using a modified processing technique. The long-term performance of the graft was evaluated by implantation in a rat abdominal aorta model. Ultrasound analysis showed that the grafts remained patent without aneurysm or stenosis occurring even after 12 months of implantation. Macroporous structure improved the cell ingrowth and thus promoted tissue regenerated at 3 months. More importantly, there was no sign of adverse remodeling, such as calcification within the graft wall after 12 months. Therefore, electrospun PCL vascular grafts with modified macroporous processing hold potential to be an artery substitute for long-term implantation.

Here, we present a modified electrospinning method to fabricate PCL vascular grafts with thick fibers and large pores, and describe a protocol to evaluate the in vivo performance in a rat model of abdominal aorta replacement.

优化结构静电纺丝血管移植在大鼠模型中的植入

Here, we present a protocol to fabricate macroporous PCL vascular graft and describe an evaluation protocol by using a rat model of abdominal aorta ...