The authors are solely responsible for the design and conduct of the presented study. Dr. Marbacher was supported by a grant from the Swiss National Science Foundation (PBSKP3-123454). The authors declare no conflict of interests. 
Author contributions to the study and manuscript preparation include the following. Conception and design: SM, JM, JF. Acquisition of data: SM, EA, JF. Analysis and interpretation of data: SM, JF, JM. Drafting the article: SM, JF, JM. Critically revising the article: JH, MN. Statistical analysis: SM, JF. Study supervision: JF, JH, MN.

注：雄性Wistar大鼠（平均体重：356±44克; 10-14周龄）被安置在动物房在22-24°C和12小时光/暗周期可免费使用颗粒饲料，普通自来水并且还获得了人文关怀，符合机构的指导方针。实验进行了审查和批准委员会动物福利在芬兰赫尔辛基大学。 注意：在下面的演示我们的手术方法如下：麻醉大鼠重量适应皮下注射盐酸美托咪定（0.5毫克/千克）和腹腔注射盐酸氯胺酮（50毫克/千克）。试验缺乏脚趾捏反射的，以确认该鼠被完全麻醉。申请眼膏，夹在外科手术部位，并用干净的合适的消毒剂的皮肤，例如氯己定，无论是在醇或水。洗净双手，穿上防护服，头罩和面掩模，并无菌手术手套。有手术助手助剂在保持无菌手术条件，并记录在手术特性（如表1所列）。通过呼吸频率，心脏速率和反应后对伤害性刺激（脚趾捏试验）监测麻醉每15分钟的手术过程中的深度。皮下注射丁丙诺啡（0.03毫克/千克）的混合物中对术后镇痛，必要时，每12小时重复。 1，硬件，耗材和定位<ol><li>养小动物手术室安静，无菌，保持室内温度在23 +/- 3℃。为了进行实验性动脉瘤手术以下最少的设备是必要的： <ol><li>使用桌面手术显微镜最好配备一个助手范围和数码显微镜摄像头。使用无孔可重复使用的工作表面和清洗的工具表面保护实验台。 </li><li&gt;准备消耗品如皮肤消毒剂，生理盐水，更小和更大的纱布拭子和缝合材料包括9-0微缝合线，以5-0不可吸收的，和3-0可吸收缝线。 </li><li>使用下列标准的手术器械：手术剪，组织钳，软组织扩张器或自保持牵开器，以及两个蚊子手术钳。 </li><li>使用碱性显微外科手术器械组，其包括：弯曲的微针夹持器，一个弯曲和两个直的microforceps和一个直的或弯曲microscissor。 </li><li>请将显微器械的肾菜充满了无菌生理盐水，以保持仪器在手术过程中不粘性和清洁。肾脏菜被填充有橡胶垫或外科手套，以防止损坏的微型仪器的尖端。确保所有物资都是无菌的过程无菌操作，根据对实验动物的生存术目前执行的建议<sup&gt; 8-10。 </li><li>此外，使用血管夹敷贴三无创伤临时血管夹。重要的是所用的夹具具有低的收功率，以防止损伤非常薄的壁的大鼠主动脉。还准备了半毫米比例尺，彩色小橡胶垫，和短钝针一把尺子。 </li><li>将大鼠在仰卧位，固定两前和后脚爪与外科胶带不施加拉伸或压缩到皮肤上，并通过将其背部的腰部区域下弯曲其背部用粗标记或烧灼笔。重要的是要获得尽可能多的腰椎前凸尽可能以改善腹膜后曝光并获得下腹主动脉便于显微外科吻合术。这种定位推荐腹主动脉瘤的创建，但不是必需的胸部移植收获。 </li></ol></li></ol> 2，嫁接收获<ol><li> ü根据一般麻醉，打开胸腔（开始移植缺血时间）。适用于有毒脚趾捏，以确认鼠是在进行以下步骤之前，反应迟钝。通过midventral腹壁切开，确定膜片只是肝上，并切断了结缔组织，在振动板的底部，以允许访问的左肋。随着大剪刀，钝面朝下，通过排骨切成只是1厘米左肋骨中线的右侧，并打开胸腔。肺是动员右侧的心脏。牺牲大鼠心内注射氯胺酮hydrochloride.Pulmonary躯干，左锁骨下静脉，左颅腔静脉过量，和奇静脉干预近端主动脉的收获。 </li><li>尾椎突出的静脉有一个很好的切入点，开始使用微型剪刀和microforceps胸降主动脉的夹层。 </li><li>跟踪胸主动脉b从胸部的背墙向上的ack对主动脉弓轻轻钝性回缩和解剖与手术蚊子夹。 </li><li>钳，然后用剪刀剪的静脉。保持对静脉的夹子和使用的牵开，露出底层的主动脉弓。 </li><li>放置一个非吸收性6-0丝线结扎只是第一肋间动脉离开主动脉以上。 </li><li>切略低于左锁骨下动脉，然后在下面的绑带降主动脉。微调可以为了得到一个垂直标准化动脉瘤几何来完成，或如果需要的话，得到的动脉瘤和主动脉的轴线之间的特定角度。测量其宽度和长度的接枝。 注：收获移植物可以立刻移植到受体鼠或进一步加工以达到接枝壁的脱细胞。脱细胞的移植物可以被存储在-4摄氏度，直到重新植入在稍后的日期（ 图1）。日动脉瘤壁电子脱细胞已被证明易患动脉瘤放大11。 </li></ol> 3，移植脱细胞<ol style="margin-left: 40px;"><li>冷冻供体移植物在磷酸盐缓冲盐水中，在-4℃。 </li><li>第二天，解冻的移植物，漂洗用纯化和去离子水在室温下，并在0.1％十二烷基硫酸钠孵育10小时，在37℃。 </li><li>最后，轻轻摇动洗十二烷基硫酸钠处理的移植物三次，重新冻结在磷酸盐缓冲盐水中，并保持在-4℃下直到使用。 </li></ol> 4，创建瘤<ol style="margin-left: 40px;"><li>腹主动脉夹层<ol><li>后的动物已经被麻醉，夹在手术部位的毛发和清洁用合适的消毒皮肤。测试缺乏前皮肤切口脚趾捏响应。启动切口1厘米靠近生殖器íÑ​​midventral位置（开始运作时）。小心地从下面的肌肉分离的皮肤。解剖完一到两厘米胸骨下方。 </li><li>仔细用力拉起下面的腹部肌肉，以避免损坏底层器官。向上延伸沿白线的长度切至剑突和在膀胱的水平结束于尾方向。 </li><li>轻轻施加压力，膀胱清空，以便于进入腹膜后间隙。 </li><li>移动小肠和盲肠突出到右侧或左侧。鉴定大肠即降结肠的腹腔的左下角。 </li><li>切在小肠和颅方向下行结肠之间的韧带，使背侧体壁的更宽的曝光。放置一个自保持牵开器，以保持大便分离。 </li><li>中端 - 侧吻合动脉瘤í的理想地点■在肾脏和髂腰静脉之间的水平找到。腹主动脉位于腹膜后嵌在脂肪组织中。解剖过程中要特别注意的配对几乎是透明的输尿管和睾丸的血管。 </li><li>如果需要的肠子进一步回缩，使用更大的纱布掉。腰椎前凸通过将烧灼笔或类似物件下的大鼠的下背部中的定位感应，显著降低肠收缩的必要性。 </li><li>背侧体壁的腹侧表面覆盖有薄的壁层腹膜。一旦这个被打开时，可视化的主动脉的正下方。在从相邻的大静脉，​​腹主动脉的小心尖锐和钝器解剖把握仅外膜，以避免损坏血管壁。 </li><li>偶尔，小腰动脉出现如从腹主动脉的背侧表面节段性容器和干扰制剂。结扎血管和切割东东DED以避免在缝合动脉逆行渗出。使用弯曲的微镊子可以便于在深度结扎位置。 </li></ol></li><li>端 - 端吻合术<ol><li>把彩色的橡胶垫腹主动脉下方，用小纱布拭装潢吧。在计划的吻合部位的水平删除疏松结缔组织和外膜。 </li><li>夹紧腹主动脉远端吻合，再近端（开始主动脉阻断时间）。这将确保该船只的坚定填充，便于后续的动脉切开术。 </li><li>执行使用是直的或弯曲的微型剪刀动脉切开术。的微型镊子举起一小块血管壁上切出一个椭圆形的。 </li><li>用生理盐水用钝尖的针两个方向彻底冲洗动脉。 </li><li>将此端 - 侧吻合术的前两个缝线在动脉切开术的近端和远端（启动吻合时间）。避免与微型镊子抓住血管壁只要有可能。确保每个缝线放置穿过容器壁的所有层。 </li><li>进行缝合无论是作为连续或间断缝合。如果中断缝合选择那么首先将背面9点钟缝合。随后的缝线可以被间隔开起相邻的第一缝合线。认真把握外膜。避免内膜任何压缩/抓。 </li><li>当后墙完成后，检查吻合术缝合错位的腔内一部分。在位于前侧的相同顺序执行相同的程序。保证在第一共三节每缝合牢固，但不要太紧。 </li></ol></li><li>止血和封闭<ol><li>之后，端侧吻合术完成（结束时间吻合），冲洗的部位用生理盐水和先移除远端夹，以便回流（完主动脉阻断时间和结束移植缺血TI我）。 </li><li>如果发生从回流的额外线圈，可能需要显着的出血（开始止血的时间）。如遇轻微渗出，达到止血用在出血部位用一小块纱布棉签轻轻按压。 </li><li>去掉近端血管钳，冲洗吻合部位一次以上，并切断绑带的剩余端部处的动脉瘤的圆顶。 </li><li>在高峰动脉脉搏波，确认动脉瘤被观察动脉瘤体积增大通畅。通过直接“挤奶测试”评估远端腹主动脉通畅。 </li><li>取出塑料片和下面的小纱布拭。所创建的动脉瘤内的脉动血液洄清晰可见。 </li><li>盖周围的小块脂肪组织或Spongostan的额外的止血吻合缝合线，如果小渗出仍然存在（止血的结束时间）。 </li><li>取下软组织扩张器和纱布棉签。将小肠，盲肠，而脂肪量早在他们的正确位置。 </li><li>关闭使用5-0，4-0，3-0，或可吸收或不可吸收缝合线，接着用连续缝合技术，用3-0可吸收缝线polyfilament（结束工作时间）皮肤伤口闭合中线腹部肌肉。注意：根据我们的经验耐受大鼠皮肤可吸收缝合线比非吸收性缝合线单丝更好。 </li></ol></li></ol>

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作者没有财务或商业利益的任何药物，材料或使用的设备。

在我们的囊状动脉瘤的复杂生物学的理解进展取决于流行病学和临床资料，在动物模型3,12,13辅以实验室检查对患者样本和实验工作分析。 小的动物，如大鼠本质上是与实验和壳体较低的成本相关联，并且减少需要专门的设备。的时间少于60分钟的大鼠的侧壁动脉瘤的显微创建的平均总操作时间比用于创建在兔子和狗2,14,15更复杂的显微静脉袋动脉分叉部动脉瘤...

囊状动脉瘤破裂引起危及生命的出血导致中风，永久性的神经损伤，甚至死亡。破裂可以通过显微外科夹闭或血管内动脉瘤栓塞预防。一种医疗治疗，以防止动脉瘤的生长和破裂尚未确立。 需要为囊状动脉瘤的实验模型，研究动脉瘤并为新的治疗设备和策略，测试生物。为了这些目的，在不同物种的几个不同的模型已经开发和发布1。在猪，狗，兔较大动脉瘤模型，优选使用在复杂的动脉瘤架构1,2测试腔内的创新。鼠动脉瘤模型，另一方面，允许在转基因物种3,4测试研究问题，并促进澄清瘤生物学中的细胞和分子水平远远大于1种更好。虽然血管内反式颈动脉和反式髂设备部署仅限于大鼠（&gt; 400-500克）和支架小于2.0毫米和1.5毫米的直径5,6，支架也可以通过直接插入置于腹主动脉段窝藏实验动脉瘤。使用大鼠显微腹主动脉侧壁动脉瘤模型先前的工作证明了其在测试新的栓塞装置的可行性和其在研究动脉瘤生物学3,7的分子基础使用。 很多目前公布的实验囊状动脉瘤模型需要昂贵的设备，特殊环境（ 如无菌手术室用透视功能），介入放射学的能力，或使用昂贵的品种。这些要求限制的广泛使用这些模式，并且导致在不同的实验室，这米使用不同的模型碟刹的数据比较和汇总分析困难的，如果不是不可能的。一个简单的，坚固的，并且便宜的实验模型是必要的，可用于在各种实验室标准化的方式，以获得来自不同机构的比较的结果，一个标准化的工具。为此，我们建立了大鼠主动脉壁囊状动脉瘤模型。 本报告的目的是介绍一步一步的过程说明，关于必要的设备，并讨论了在大鼠腹主动脉旁瘤显微外科成功创造重要的解剖及手术特点。

<table border="0" cellpadding="0" cellspacing="0" width="752">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:231px;">Name of Material/ Equipment</td>
			<td style="width:236px;">Company</td>
			<td style="width:119px;">Catalog Number</td>
			<td style="width:167px;">Comments/Description</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:231px;">Medetomidine</td>
			<td style="width:236px;"></td>
			<td style="width:119px;"></td>
			<td>Any genericon</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:231px;">Ketamin</td>
			<td style="width:236px;"></td>
			<td style="width:119px;"></td>
			<td>Any genericon</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:231px;">Buprenorphine</td>
			<td style="width:236px;"></td>
			<td style="width:119px;"></td>
			<td>Any genericon</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:231px;">Phosphate buffered saline</td>
			<td style="width:236px;"></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:231px;">Sodium dodecyl sulfate (0.1%)</td>
			<td style="width:236px;"></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:231px;">3-0 resorbable suture</td>
			<td style="width:236px;">Ethicon Inc., USA</td>
			<td><a name="RANGE!C7">VCP824G</a></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:231px;">5-0 non absorbable suture</td>
			<td style="width:236px;">Ethicon Inc., USA</td>
			<td>8618G</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:231px;">6-0 non absorbable silk suture</td>
			<td style="width:236px;">B. Braun, Germany</td>
			<td style="width:119px;">C0761060</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:231px;">9-0 nylon micro suture</td>
			<td style="width:236px;">B. Braun, Germany</td>
			<td style="width:119px;">G1118471</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:231px;">Spongostan</td>
			<td style="width:236px;">Ethicon Inc., USA</td>
			<td style="width:119px;">MS0002</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:231px;">Operation microscope</td>
			<td style="width:236px;">Leica , Germany</td>
			<td style="width:119px;">M651</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:231px;">Digital microscope camera</td>
			<td style="width:236px;">Sony, Japan</td>
			<td style="width:119px;">SSC-DC58AP</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:231px;">Standard surgical instruments</td>
			<td style="width:236px;">B. Braun, Germany</td>
			<td style="width:119px;">Multiple</td>
			<td>See protocol 1.4</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:231px;">Microsurgical instruments</td>
			<td style="width:236px;">B. Braun, Germany</td>
			<td style="width:119px;">Multiple</td>
			<td>See protocol 1.5</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:231px;">Vascular clip applicator</td>
			<td style="width:236px;">B. Braun, Germany</td>
			<td style="width:119px;">FT495T</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:231px;">Temporary vascular clamps</td>
			<td style="width:236px;">B. Braun, Germany</td>
			<td style="width:119px;">FT250T</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;">Graph Pad Prism statistical software&#160;</td>
			<td style="width:236px;">GraphPad Software, San Diego, California, USA</td>
			<td style="width:119px;"></td>
			<td>V 6.02 for Windows</td>
		</tr>
	</tbody>
</table>

the helsinki rat microsurgical sidewall aneurysm model

实验囊状动脉瘤模型所必需的试验新型手术和腔内治疗方案和设备之前，它们被引入到临床实践。此外，需要实验模型以阐明复杂动脉瘤生物学导致破裂囊状动脉瘤的。 几种不同类型的实验模型的囊状动脉瘤都建立了不同的物种。他们中的许多，但是，需要特殊的技能，昂贵的设备，或特殊的环境中，这限制了它们的广泛使用。一个简单的，强大的，廉价的实验模型是需要的，可以在各种机构的标准化方式使用标准化的工具。 显微大鼠腹主动脉旁瘤模型结合来研究这两个新的腔内治疗策略和动脉瘤生物学在一个标准化的和廉价的分子基础的可能性的方式。标准化的移植物由形状，大小和几何形状的装置被从供体大鼠的胸降主动脉收获，然后移植到同系受体鼠。动脉瘤是缝合的端至端具有连续或间断的9-0尼龙缝线的下腹主动脉。 我们目前对必要的设备一步一步的程序指令，信息，讨论重要的解剖和手术细节在大鼠腹主动脉旁瘤的显微外科成功创建。

飞行员系列包括14只。随后，共84动物根据协议提出的三月和九月之间的几个研究项目2012年增加29个动物作为供体的囊状动脉移植手术了。剩余的实验使用收获并从用大鼠同性别，应变，体重和年龄的先前实验中存储经预处理的移植物进行的。 体重，总体运行时间，主动脉阻断时间，时间吻合的创作，时间以止血吻合的创建，接枝缺血时间后，与动脉瘤的尺寸，在创建时（动?...

Watch this Scientific Journal Video about The Helsinki Rat Microsurgical Sidewall Aneurysm Model at JoVE.com

The Helsinki Rat Microsurgical Sidewall Aneurysm Model

Microsurgical sidewall aneurysms in rats are created by end-to-side anastomosis of an aortic graft to the abdominal aorta. We present step-by-step instructions and discuss anatomical and surgical details for successful experimental saccular aneurysm creation.

赫尔辛基鼠显微旁瘤模型

Experimental saccular aneurysm models are necessary for testing novel surgical and endovascular treatment options and devices before they are introduced ...

the-helsinki-rat-microsurgical-sidewall-aneurysm-model

Research

JoVE Journal

Medicine

12.1K 次观看.  University of Helsinki, Helsinki, Finland. Microsurgical sidewall aneurysms in rats are created by end-to-side anastomosis of an aortic graft to the abdominal aorta. We present step-by-step instructions and discuss anatomical and surgical details for successful experimental saccular aneurysm creation.

视频: The Helsinki Rat Microsurgical Sidewall Aneurysm Model

Microsurgical Venous Pouch Arterial-Bifurcation Aneurysms in the Rabbit Model: Technical Aspects

For ruptured human cerebral aneurysms endovascular embolization has become an equivalent alternative to aneurysm clipping.1 However, large clinical trials have shown disappointing long-term results with unacceptable high rates of aneurysm recanalization and delayed aneurysm rupture.2 To overcome these problems, animal experimental studies are crucial for the development of better endovascular devices.3-5
Several animal models in rats, rabbits, canines and swine are available.6-8 Comparisons of the different animal models showed the superiority of the rabbit model with regard to hemodynamics and comparability of the coagulation system and cost-effectiveness.9-11 
The venous pouch arterial bifurcation model in rabbits is formed by a venous pouch sutured into an artificially created true bifurcation of both common carotid arteries (CCA). The main advantage of this model are true bifurcational hemodynamics.12 The major drawbacks are the sofar high microsurgical technical demands and high morbidity and mortality rates of up to 50%.13 These limitations have resulted in less frequent use of this aneurysm model in the recent years. These shortcomings could be overcome with improved surgical procedures and modified peri- and postoperative analgetic management and anticoagulation.14-16 Our techniques reported in this paper demonstrate this optimized technique for microsurgical creation of arterial bifurcation aneurysms.

An optimized technique for the microsurgical creation of arterial bifurcation aneurysms mimicking bifurcation human cerebral aneurysms is described. A venous pouch is sutured into an artificially created true bifurcation of both common carotid arteries. Facilitated microsurgical techniques and aggressive postoperative anticoagulation and analgesia lead to minimized morbidity rates and high aneurysm patency rates.

显微静脉袋分岔动脉瘤在兔模型：技术问题

For ruptured human cerebral aneurysms endovascular embolization has become an equivalent alternative to aneurysm clipping.1 However, large clinical trials ...

科研

医学

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 ...

Demonstration of the Rat Ischemic Skin Wound Model

The propensity for chronic wounds in humans increases with ageing, disease conditions such as diabetes and impaired cardiovascular function, and unrelieved pressure due to immobility. Animal models have been developed that attempt to mimic these conditions for the purpose of furthering our understanding of the complexity of chronic wounds. The model described herein is a rat ischemic skin flap model that permits a prolonged reduction of blood flow resulting in wounds that become ischemic and resemble a chronic wound phenotype (reduced vascularization, increased inflammation and delayed wound closure). It consists of a bipedicled dorsal flap with 2 ischemic wounds placed centrally and 2 non-ischemic wounds lateral to the flap as controls. A novel addition to this ischemic skin flap model is the placement of a silicone sheet beneath the flap that functions as a barrier and a splint to prevent revascularization and reduce contraction as the wounds heal. Despite the debate of using rats for wound healing studies due to their quite distinct anatomic and physiologic differences compared to humans (i.e., the presence of a panniculus carnosus muscle, short life-span, increased number of hair follicles, and their ability to heal infected wounds) the modifications employed in this model make it a valuable alternative to previously developed ischemic skin flap models.

The rat, due to its size, availability, and rather docile behavior, has been utilized as a research model for many years. The goal of this protocol is to utilize the rat as an ischemic skin wound healing model to provide valuable insight into the pathophysiology of chronic wounds.

在大鼠缺血性皮肤创伤模型的演示

The propensity for chronic wounds in humans increases with ageing, disease conditions such as diabetes and impaired cardiovascular function, and ...

The Dimethylnitrosamine Induced Liver Fibrosis Model in the Rat

Four to six week old, male Wistar rats were used to produce animal models of liver fibrosis. The process requires four weeks of administration of 10 mg/kg dimethylnitrosamine (DMN), given intraperitoneally for three consecutive days per week. Intraperitoneal injections were performed in the fume hood as DMN is a known hepatoxin and carcinogen. The model has several advantages. Firstly, liver changes can be studied sequentially or at particular stages of interest. Secondly, the stage of liver disease can be monitored by measurement of serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) enzymes. Thirdly, the severity of liver damage at different stages can be confirmed by sacrifice of animals at designated time points, followed by histological examination of Masson&#39;s Trichome stained liver tissues. After four weeks of DMN dosing, the typical fibrosis score is 5 to 6 on the Ishak scale. The model can be reproduced consistently and has been widely used to assess the efficacy of potential anti-fibrotic agents.

We describe a method to produce an animal model of liver fibrosis in the rat, and assess the degree of fibrosis by histological examination of the liver. The model can be used to study the development of liver disease as well as to test the efficacy of potential anti-fibrotic agents.

该二甲基亚硝胺诱发肝纤维化模型大鼠

Four to six week old, male Wistar rats were used to produce animal models of liver fibrosis. The process requires four weeks of administration of 10 mg/kg ...

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 ...

Corneal Epithelial Abrasion with Ocular Burr As a Model for Cornea Wound Healing

The murine cornea provides an excellent model to study wound healing. The cornea is the outermost layer of the eye, and thus is the first defense to injury. In fact, the most common type of eye injury found in clinic is a corneal abrasion. Here, we utilize an ocular burr to induce an abrasion resulting in removal of the corneal epithelium in vivo on anesthetized mice. This method allows for targeted and reproducible epithelial disruption, leaving other areas intact. In addition, we describe the visualization of the abraded epithelium with fluorescein staining and provide concrete advice on how to visualize the abraded cornea. Then, we follow the timeline of wound healing 0, 18, and 72 h after abrasion, until the wound is re-epithelialized. The epithelial abrasion model of corneal injury is ideal for studies on epithelial cell proliferation, migration and re-epithelialization of the corneal layers. However, this method is not optimal to study stromal activation during wound healing, because the ocular burr does not penetrate to the stromal cell layers. This method is also suitable for clinical applications, for example, pre-clinical test of drug effectiveness.

This protocol describes a method to inflict an abrasion to the ocular surface of the mouse, and to follow the wound healing process thereafter. The protocol takes advantage of an ocular burr to partially remove the surface epithelium of the eye in anaesthetized mice.

眼毛刺角膜上皮磨损与角膜创面愈合的模型研究

The murine cornea provides an excellent model to study wound healing. The cornea is the outermost layer of the eye, and thus is the first defense to ...

An Ex Vivo Tissue Culture Model for Fibrovascular Complications in Proliferative Diabetic Retinopathy

Diabetic retinopathy (DR) is the most common microvascular complication of diabetes and one of the leading causes of blindness in working-age adults. No current animal models of diabetes and oxygen-induced retinopathy develop the full-range progressive changes manifested in human proliferative diabetic retinopathy (PDR). Therefore, understanding of the disease pathogenesis and pathophysiology has relied largely on the use of histological sections and vitreous samples in approaches that only provide steady-state information on the involved pathogenic factors. Increasing evidence indicates that dynamic cell-cell and cell-extracellular matrix (ECM) interactions in the context of three-dimensional (3D) microenvironments are essential for the mechanistic and functional studies towards the development of new treatment strategies. Therefore, we hypothesized that the pathological fibrovascular tissue surgically excised from eyes with PDR could be utilized to reliably unravel the cellular and molecular mechanisms of this devastating disease and to test the potential for novel clinical interventions. Towards this end, we developed a novel method for 3D ex vivo culture of surgically-excised patient-derived fibrovascular tissue (FT), which will serve as a relevant model of human PDR pathophysiology. The FTs are dissected into explants and embedded in fibrin matrix for ex vivo culture and 3D characterization. Whole-mount immunofluorescence of the native FTs and end-point cultures allows thorough investigation of tissue composition and multicellular processes, highlighting the importance of 3D tissue-level characterization for uncovering relevant features of PDR pathophysiology. This model will allow the simultaneous assessment of molecular mechanisms, cellular/tissue processes and treatment responses in the complex context of dynamic biochemical and physical interactions within the PDR tissue architecture and microenvironment. Since this model recapitulates PDR pathophysiology, it will also be amenable for testing or developing new treatments.

Here, we present a protocol to study the pathophysiology of proliferative diabetic retinopathy by using patient-derived, surgically-excised, fibrovascular tissues for three-dimensional native tissue characterization and ex vivo culture. This ex vivo culture model is also amenable for testing or developing new treatments.

增殖性糖尿病视网膜病变纤维血管并发症的体外组织培养模型

Diabetic retinopathy (DR) is the most common microvascular complication of diabetes and one of the leading causes of blindness in working-age adults. No ...

Murine Surgical Model of Topical Elastase Induced Descending Thoracic Aortic Aneurysm

According to the Center for Disease Control, aortic aneurysms (AAs) were considered a leading cause of death in all races and both sexes from 1999-2016. An aneurysm forms as a result of progressive weakening and eventual dilation of the aorta, which can rupture or tear once it reaches a critical diameter. Aneurysms of the descending aorta in the chest, called descending thoracic aortic aneurysms (dTAA), make up a large proportion of aneurysm cases in the United States. Uncontained dTAA rupture is almost universally lethal, and elective repair has a high rate of morbidity and mortality. The purpose of our model is to study dTAA specifically, to elucidate the pathophysiology of dTAA and to search for molecular targets to halt the growth or reduce the size of dTAA. By having a murine model to study thoracic pathology precisely, targeted therapies can be developed to specifically test dTAA. The method is based on the placement of porcine pancreatic elastase (PPE) directly on the outer murine aortic wall after surgical exposure. This creates a destructive and inflammatory reaction, which weakens the aortic wall and allows for aneurysm formation over weeks to months. Though murine models possess limitations, our dTAA model produces robust aneurysms of predictable size. Furthermore, this model can be used to test genetic and pharmaceutical targets which may arrest dTAA growth or prevent rupture. In human patients, interventions such as these could help avoid aneurysm rupture, and difficult surgical intervention.

We describe a surgical protocol to consistently induce robust descending thoracic aortic aneurysms in mice. The procedure involves left thoracotomy, thoracic aorta exposure, and placement of a sponge soaked in porcine pancreatic elastase on the aortic wall.

局部性乳酸酶诱导下降胸腔主动脉瘤的毛尿外科模型

According to the Center for Disease Control, aortic aneurysms (AAs) were considered a leading cause of death in all races and both sexes from 1999-2016. ...

Porcine Model of Infrarenal Abdominal Aortic Aneurysm

Large animal models to study abdominal aortic aneurysms are sparse. The purpose of this model is to create reproducible, clinically significant infrarenal abdominal aortic aneurysms (AAA) in swine. To achieve this, we use a combination of balloon angioplasty, elastase and collagenase, and a lysyl oxidase inhibitor, called &#946;-aminopropionitrile (BAPN), to create clinically significant infrarenal aortic aneurysms, analogous to human disease.
Noncastrated male swine are fed BAPN for 7 days prior to surgery to achieve a steady state in the blood. A midline laparotomy is performed and the infrarenal aorta is circumferentially dissected. An initial measurement is recorded prior to aneurysm induction with a combination of balloon angioplasty, elastase (500 units)/collagenase (8000 units) perfusion, and topical elastase application. Swine are fed BAPN daily until terminal procedure on either postoperative day 7, 14, or 28, at which time the aneurysm is measured, and tissue procured. BAPN + surgery pigs are compared to pigs that underwent surgery alone.
Swine treated with BAPN and surgery had a mean aortic dilation of 89.9% &#177; 47.4% at day 7, 105.4% &#177; 58.1% at day 14, and 113.5% &#177; 30.2% at day 28. Pigs treated with surgery alone had significantly smaller aneurysms compared to BAPN + surgery animals at day 28 (p &#60; 0.0003). The BAPN + surgery group had macroscopic and immunohistochemical evidence of end stage aneurysmal disease.
Clinically significant infrarenal AAA can be induced using balloon angioplasty, elastase/collagenase perfusion and topical application, supplemented with oral BAPN. This model creates large, clinically significant AAA with hallmarks of human disease. This has important implications for the elucidation of AAA pathogenesis and testing of novel therapies and devices for the treatment of AAA. Limitations of the model include variation in BAPN ingested by swine, quality of elastase perfusion, and cost of BAPN.&#160;

This novel model creates robust infrarenal abdominal aortic aneurysms in swine using a combination of balloon angioplasty, elastase/collagenase perfusion, topical elastase application, and oral compound &#946;-aminopropionitrile administration, which interferes with collagen cross-linking.

肾上腺腹动脉瘤的猪模型

Large animal models to study abdominal aortic aneurysms are sparse. The purpose of this model is to create reproducible, clinically significant infrarenal ...

Vessel-Sparing Microsurgical Longitudinal Intussusception Vasoepididymostomy to Treat Epididymal Obstructive Azoospermia

The epididymis is a common site of obstruction in obstructive azoospermia (OA). Vasoepididymostomy has become an important method for the treatment of epididymal OA since 2000. There are two challenges in classic microscopic vasoepididymostomy. First, anastomosis of the vas deferens and epididymis is performed with double-needle sutures. However, there is a lack of good-quality and cost-effective double-needle sutures in China, which leads to increased difficulty and poor success rates of anastomosis. Second, the separation of the vas deferens does not retain vasculature, although the vas deferens vasculature plays an important role in the blood supply to the vas deferens, epididymis, and testis. This affects the blood supply to the anastomotic area and epididymis.
Therefore, this team has made innovative improvements to address these problems. Good-quality, cost-effective, single-needle sutures, which are easy to purchase in China and other countries, were used in microsurgical longitudinal intussusception vasoepididymostomy. This can optimize the operation procedure and shorten the operation time while ensuring the success rate of the anastomosis. The surgical method of preserving the vas deferens vessels was innovatively proposed because the etiology of epididymal OA is mostly inflammatory in China. The protection of the blood supply to the vas deferens and epididymis is maximized using microsurgical forceps to separate and protect the vasculature. Patency reached 81.7% in the postoperative follow-up, indicating a better surgical treatment effect.

The present protocol describes a vessel-sparing, longitudinal intussusception vasoepididymostomy using readily available single-needle sutures in China, a safe and effective procedure, which may improve patients' patency and natural pregnancy rates.

血管保留性显微外科纵向套叠血管表皮造口术治疗附睾梗阻性无精子症

The epididymis is a common site of obstruction in obstructive azoospermia (OA). Vasoepididymostomy has become an important method for the treatment of ...