这个工作是由医学研究委员会授予G1000299。 相应的作者要感谢协助在手术过程中，爱丁堡大学的加里·博思威克。 笔者想从海伦·道格拉斯和麦荣恩确认意见，并允许我们观察到其深腹壁（DIEP）皮瓣法（Canniesburn整形外科单位，格拉斯哥皇家医院，84城堡街G4 0SF，格拉斯哥，英国）。 作者还要感谢他的帮助下对这篇文章的视频在爱丁堡大学的加里·布莱基。

所有手术是按照由英国内政部和英国爱丁堡大学的兽医服务局的指引载。 1。手术程序集的说明<ol><li>转变成干净的手术袍，长袍，磨砂帽和口罩。清洁所有表面手术室，包括70％的异丙醇用2％洗必泰设备。 </li><li>手术前，高压灭菌器，将用于在程序中的所有手术用品和仪器。每次手术的无菌包应包括：窗帘，纱布，棉签涂药，矽胶片和手术器械，请参阅表的具体的手术材料和工具，和图1。大鼠称重和测量适当体积的丁丙诺啡（0.04毫克/千克），以皮下注射前1小时完成的过程。布局; 3×1毫升注射器皮下积液管理外科学每2×6-0薇乔缝合线，1×5-0 Ethilon缝合线，无菌的标记与标尺和10刀片一次性解剖刀，4 - 5对无菌手套，和一个手持式的烧灼部。 </li><li>将4×10毫升无菌0.9％盐水小瓶在水浴中加热到37℃。这将用来更换皮下积液（1毫升/公斤/小时）和冲洗手术部位。设置毯的homeothermic，直肠探头，热灯，手术显微镜和麻醉钻机。打开激光及其软件，设置另一个麻醉钻机和下方的黑色垫子，动物会躺在扫描过程中放置​​散热垫。 </li><li>使用男，Lewis大鼠，体重250-300克。 7天自由采食和饮水，12小时明暗周期之前，任何手术与食品家鼠进行。 </li></ol> 2。麻醉和皮肤准备<ol><li>将大鼠麻醉钻机的感应室和管理1L/min时O 2与4％的异氟醚2-3分钟诱导麻醉。从箱内取出麻醉大鼠仰卧放置在干净，加热垫。维持异氟醚1.5％使用鼻锥。应用lacrilube或类似剂，以防止在手术过程中角膜擦伤。进行足垫捏测试，以确保动物有足够的麻醉，然后再进行。重复这最后的测试，在每个过程中的重要一步，并相应地调整吸入麻醉药浓度。 </li><li>密切剃腹前壁使用的电动剃须刀，使整个腹部的表面暴露。应用脱毛霜由供应商建议的时间。取出奶油和温暖无菌生理盐水彻底冲洗皮肤，删除所有痕迹的奶油。应用2％洗必泰，70％异丙醇对皮肤和晾干，然后再继续。这是标准的皮肤准备在我们单位，根据目前外科手术部位感染的证据。34请ð与您的兽医部iscuss标准是什么程序在你的单元选出前皮肤准备协议。 </li><li>广场两侧的窗帘大鼠和照顾，让他们无菌。戴上无菌手套和一个助手的帮助下打开无菌包。将所有的仪器在一个悬垂的缝合线，纱布，棉花涂药，矽胶片和无菌记号笔与其他统治者。 </li><li>确定中线剑突和尾部作为参考点。标记中线。测量0.8厘米剑突下方，标志着这一点。画一条线垂直于从该点向中线。以中线襟翼标记为中心出1厘米和2厘米的左侧和中线的右侧。绘制从点向中线平行的垂直线。原来的水平线上测量4厘米以下，并绘制另一个平行。按照这些说明一个4×4厘米的皮瓣分为4个相等条划定（参见图2）。 </li></ol> 3。激光多普勒成像<ol><li>小心地将大鼠LDI扫描仪第二麻醉钻机在鼻锥。继续麻醉1.5％异氟醚，1L/min时O 2。打开激光，并按照制造商的指示，开始扫描。保存扫描文件后返回大鼠第一钻机和重新插入直肠探头的homeothermic毯采用软白石蜡作为润滑剂。 </li></ol> 4。原位 TRAM皮瓣-肌IRI模型<ol><li>再擦洗双手把新鲜无菌手套。剪出一个直径5厘米的圆圈的中心，其余无菌的悬垂性和放置在裸露腹部创造一个搭着无菌场。 </li><li>做一个切口沿左路横向标记边缘（ 图3A和B）。达到止血效果。类似切口上下水平线中线的左侧。达到止血效果。 </li><li>脂肪覆左下腹直肌鞘应该是可见的。仔细使用镊子和精美的虹膜剪下面这块肥肉。小心不要损坏打孔器通过左，腹直肌鞘前壁。飞机开辟等夹层，立即前腹壁筋膜以上。在这架飞机周围划定边界继续解剖。在左髂窝在于大，旋髂浅血管，这些可以被捆绑或烧灼。然后延长夹层内侧谨慎，只有尽可能左腹直肌外侧缘。有一个明显的颜色变化，在这一点上，从粉红色到近白色（ 图3C）。小心灌溉面积用无菌生理盐水和检查，止血前已达到湿润的纱布放置在该区域。 </li><li>对侧上重复此过程，但这个时间延长吨Ø 白线 （中线）。小心识别和烧灼在右腹直肌（ 图3D）的中心肌产生的打孔器。如果不这样做正确术后血肿和可靠的结果，它会导致。同样达到止血，浇水湿润的纱布放置募集瓣。 </li><li>回到左前腹直肌下缘（ 图3E及F）。烧灼见过的最下穿孔。继续腹直肌鞘前壁使用microscissors的切开一个小窗口中（约0.6厘米×0.6厘米），并指出弯曲Graeffe的镊子。钝性解剖的外侧缘肌肉慢慢放下，直到肌肉变薄，但在此之前被违反后鞘。然后钳旋转，直到腹部的肌肉之上的弧形边缘的镊子，并提示是免费的内侧缘，钝性解剖内侧。饲料近似LY 6厘米5-0 Ethilon到钳口钳和配合下直肌鞘。这一步完成后，被孤立在一个占主导地位的船舶深优越的上腹血管皮瓣。盖上湿润的纱布。 </li><li>切矽胶片成椭圆用平滑拐角（ 图3G）。这些应该是大到足以覆盖大部分的面积暴露下的筋膜皮瓣部分。然而，必须谨慎，以确保可以封闭在无张力状态和内侧的椭圆形曲线的皮肤边缘可能必须通过其它穿孔机，以防止它损害流动配对。然后，将它们缝合的地方用6-0薇乔（ 图3H）。盖上湿润的纱布。 </li><li>使用简单的中断5-0 Ethilon缝合皮瓣，以减少热量和水分损失（ 图3I）挂出&#39;。盖上湿润的纱布。 </li><li>优伤口延伸到左侧的剑突（ 如图3I）。缝合当前左上象限，以提高该字段的视图。 </li><li>切去任何覆脂肪揭示优越，左，前腹直肌鞘。切开一个小窗口（0.6厘米×0.6厘米）在此鞘（ 图3J）。内侧的伤口延长，直至肌纤维轨迹的变化从垂直到看到斜肌和松散的纤维排列紧密的一致性。 </li><li>插入这两块肌肉之间仔细弯钳钝性分离，并开辟了飞机。小心砍掉尽可能优异的表面，这些弯钳切割，左腹直肌揭示底层深，腹壁上动脉和静脉（ 图3K）通过腹部。 </li><li>使用微型仪器和高功率的手术显微镜，仔细分离动脉和静脉和去掉所有周围脂肪的。 </li><li>将无创伤阿克兰钳位到一个的rtery和静脉（B-1，“V”型），并开始计时，倒计时30分钟缺血期。灌溉夹紧蒂，用纱布覆盖。我们没有雇用船只的扩张器如维拉帕米或pabavarine的但血管痉挛应该是一个问题，这些药物应予以考虑。 </li><li>辖的丁丙诺非（0.04毫克/千克）和温热的，无菌盐水中（1毫升/公斤/小时）。 </li><li>从左上角开始，停止与6-0薇乔皮下的缝线打结在剑突缝合皮瓣。 </li><li>当30分钟缺血时间已经过去，小心地取出夹具和温热的盐水灌溉蒂。检查流量已重新建立。请注意，缺血时间，这是由英国内政部权威规定。在其他部门工作的研究人员或许能够延长这一时间。扩展缺血时间将有可能导致较差的临床结果。 </li><li>的腹直肌缝合切缘用6-0薇乔到位。小心不要施加太大的紧张，因为这可能会导致扭结的船只。 </li><li>填写皮下缝合照顾到所有结埋在皮肤下面（ 图3K）。 </li><li>清洁伤口区域，并让其干燥。重新绘制瓣上的区域。 </li><li>重新扫描动物获得术后影像。 </li><li>重新申请lacrilube动物的眼睛和一个温暖的孵化器在（37°C）1小时恢复返回牧单元之前。 </li></ol>在协议的关键步骤关键的程序是确定深，优越的上腹血管。这清楚地显示在随附膜。简单地说，一个窗口被切断腹直肌鞘前壁，以暴露下面的肌肉纤维的纵向上。通过扩展腹直肌前鞘的表面解剖内侧肌纤维的变化轨迹观察fROM纵向斜。插入钝止的，弯曲的，Graeffe钳（或类似），在这两个肌纤维束的交叉点。钝性解剖横向。降不下来，使用微型剪刀，在此平面之间的肌纤维束的弯钳的上表面上。在去除Graeffe钳深，腹壁上动脉和静脉将观察腹直肌体的中点。剥去脂肪覆船只使用微型工具和应用夹具。 大鼠TRAM皮瓣，筋膜部分薄到足以允许皮瓣作为全厚皮片。为了防止这种情况，以确保这是一个真实的模型，IRI薄，灵活的硅钢片置于下方的筋膜皮瓣部分。35这一步已通过其他研究人员进行大鼠TRAM模型。17,21,25 大鼠咀嚼THROUGH节，以便确保所有缝线皮下，被埋没节。在报告的其他研究人员开展细致的缝合autocannabilism，襟翼，可避免24 继降低维护管理的丁丙诺啡麻醉1％异氟醚（1L/min时O 2）。

我们没有披露。

修改和故障排除这里介绍的协议看到游离组织移植在实验系统中再现的IRI能够进一步了解这一进程，并提供了一​​种方法，调查手段，改善IRI和改善结果。这可以很容易地修改，以产生更严重的伤害，如果它的基础上不占优势，深，腹壁下带蒂或如果缺血时间增加。 技术的限制对大鼠腹前壁有显著比大多数女性接受TRAM皮瓣乳房重建手?...

这个协议的目的是为了展示一个可靠和可重复的模型，观察缺血再灌注损伤中游离组织移植，使介入策略进行调查。 免费组织转移被定义为一个孤立的组织块，然后由该组织自体移植吻合瓣的横断本地船只船只接收站点的血管支队。该过程被称为FTT和组织受让方简称为游离皮瓣。 游离组织移植的金标准方法用于矫正复杂的，复合的缺陷地方选择不适合或无法使用。1-4缺血再灌注损伤（IRI）是不可避免的游离组织移植，有助于襟翼失效5,6及无有效的治疗。游离皮瓣手术选修性质，允许管理pharmacologi卡尔剂对IRI的前提。 IRI结果在流经受损内皮细胞活化微循环和代谢功能障碍，毛细血管通透性增加和随后的间质水肿，炎性细胞的大量涌入，8炎性介质的释放，活性氧和补体沉积。10这个复杂的过程中缺氧和随后的再灌注损伤，最终导致细胞死亡。肌IRI模型，使预处理策略，对临床结果的有效性进行评估。最近的工作作为替代人类IRI IRI研究的动物模型验证了使用通过比较分子人类受试者中观察到的变化和现有的动物数据。10,11 大鼠横向腹直肌肌皮瓣（TRAM）于1987年在德国的12，并于1993年首次被描述13英语。这种模式得到广泛普及13-25 TRAM皮瓣的基础上深，下unipedicled 14,17-22这些研究大部分被设计成，作为一种廉价的，可靠的模型来研究不同的策略，以降低游离组织移植IRI。上腹血管蒂15-18,20-22从这些研究的数据比较复杂，通过使用不同大小的皮岛（10.5 - 30厘米2）和不同长度的术后随访（2 - 10天）。在这些研究的控制臂的平均百分比总面积皮瓣坏死是69±6.2％（平均值±SEM）。应当注意，这六个文件全部采用腹直肌血管蒂的载体，但不公开，分裂和microanastomose或钳血管。 Zhang 等 23所描述的真正的，自由的大鼠TRAM皮瓣，腹壁上血管的基础上的f圈提出，船舶分为肌皮瓣转移腹股沟血管microanastomosed的。这个技术难度要求microanastomosis为0.45 - 0.5毫米口径的容器。只有15和67％，这些幸存下来。23 Zhang 等人描述的模型23是人类的自由TRAM皮瓣一个很好的模型，因为它真实地反映了FTT过程中产生伤害。大鼠TRAM皮瓣的其他已发表的模型，更准确地反映带蒂TRAM在人体产生伤害，但并不能准确地反映这些瓣IRI不接受血管蒂从来没有夹紧或划分和缺血再灌注期间执行microanastomosis。该协议和视频描述IRI被复制使用microclamps的使用大鼠TRAM游离组织移植的新模式。这更忠实地复制IRI比蒂TRAM前辈的，但在技术上更容易比performi纳克microanastomosis。通过移植的研究人员已被广泛采用microclamps重新创建IRI与实体器官移植; 26-33然而，这是第一次已经描述了在大鼠TRAM皮瓣。

<table border="1">
 <tbody>
 <tr>
 <td>Name of Reagent/Material</td>
 <td>Company</td>
 <td>Catalog Number</td>
 <td>Comments</td>
 </tr>
 <tr>
 <td>Moor LD12 laser doppler imaging scanner</td>
 <td></td>
 <td></td>
 <td><a href="http://gb.moor.co.uk/product/moorldi2-laser-doppler-imager/8" target="_blank">http://gb.moor.co.uk/product/moorldi2-laser-doppler-imager/8</a></td>
 </tr>
 <tr>
 <td>Complete homeothermic blanket system with flexible probe. Small. 230 VAC, 50 Hz</td>
 <td></td>
 <td>507221F</td>
 <td><a href="http://www.harvardapparatus.com" target="_blank">www.harvardapparatus.com</a></td>
 </tr>
 <tr>
 <td>Graeffe forceps 0.8 mm tips curved</td>
 <td></td>
 <td>11052-10</td>
 <td>2, <a href="http://www.finescience.de" target="_blank">http://www.finescience.de</a></td>
 </tr>
 <tr>
 <td>Acland clamps</td>
 <td></td>
 <td>00398 V</td>
 <td>B-1 'V' pattern clamps used on both artery and vein. <a href="http://www.merciansurgical.com/acland-clamps.pdf" target="_blank">http://www.merciansurgical.com/acland-clamps.pdf</a></td>
 </tr>
 <tr>
 <td>Clamp applicator</td>
 <td></td>
 <td>CAF-4</td>
 <td><a href="http://www.merciansurgical.com/acland-clamps.pdf" target="_blank">http://www.merciansurgical.com/acland-clamps.pdf</a></td>
 </tr>
 <tr>
 <td>Gemini cautery unit</td>
 <td></td>
 <td>726067</td>
 <td><a href="www.harvardapparatus.com" target="_blank">www.harvardapparatus.com</a></td>
 </tr>
 <tr>
 <td>Micro-vessel dilators 11 cm 0.3 mm tips 00124</td>
 <td></td>
 <td>D-5a.2</td>
 <td><a href="http://www.merciansurgical.com" target="_blank">http://www.merciansurgical.com</a></td>
 </tr>
 <tr>
 <td>Micro Jewellers Forceps 11cm angulated 00109</td>
 <td></td>
 <td>JFA-5b</td>
 <td><a href="http://www.merciansurgical.com" target="_blank">http://www.merciansurgical.com</a></td>
 </tr>
 <tr>
 <td>Micro Jewellers Forceps 11 cm straight 00108</td>
 <td></td>
 <td>JF-5</td>
 <td><a href="http://www.merciansurgical.com" target="_blank">http://www.merciansurgical.com</a></td>
 </tr>
 <tr>
 <td>Acland Single Clamps B-1V (Pair)</td>
 <td></td>
 <td>396</td>
 <td><a href="http://www.merciansurgical.com" target="_blank">http://www.merciansurgical.com</a></td>
 </tr>
 <tr>
 <td>Micro Scissors Round Handles 15 cm Straight</td>
 <td></td>
 <td>67</td>
 <td><a href="http://www.merciansurgical.com" target="_blank">http://www.merciansurgical.com</a></td>
 </tr>
 <tr>
 <td>Iris Scissors 11.5 cm Curves EASY-CUT</td>
 <td></td>
 <td>EA7613-11</td>
 <td><a href="http://www.merciansurgical.com" target="_blank">http://www.merciansurgical.com</a></td>
 </tr>
 <tr>
 <td>Mayo Scissors 14 cm Straight Chamfered Blades EASY-CUT</td>
 <td></td>
 <td>EA7652-14</td>
 <td><a href="http://www.merciansurgical.com" target="_blank">http://www.merciansurgical.com</a></td>
 </tr>
 <tr>
 <td>Derf Needle Holders 12 cm TC</td>
 <td></td>
 <td>703DE12</td>
 <td><a href="http://www.merciansurgical.com" target="_blank">http://www.merciansurgical.com</a></td>
 </tr>
 <tr>
 <td>Ethilon 5-0</td>
 <td></td>
 <td>W1618</td>
 <td><a href="http://www.farlamedical.co.uk/" target="_blank">http://www.farlamedical.co.uk/</a></td>
 </tr>
 <tr>
 <td>Vicryl rapide 6-0</td>
 <td></td>
 <td>W9913</td>
 <td><a href="http://www.millermedicalsupplies.com/" target="_blank">http://www.millermedicalsupplies.com/</a></td>
 </tr>
 <tr>
 <td>Instrapac - Adson Toothed Forceps (Extra Fine)</td>
 <td></td>
 <td>7973</td>
 <td><a href="http://www.millermedicalsupplies.com/" target="_blank">http://www.millermedicalsupplies.com/</a></td>
 </tr>
 <tr>
 <td>Castroviejo needle holders</td>
 <td></td>
 <td>12565-14</td>
 <td><a href="http://s-and-t.ne" target="_blank">http://s-and-t.ne</a></td>
 </tr>
 <tr>
 <td>Heat Lamp</td>
 <td></td>
 <td></td>
 <td><a href="http://www.chicken-house.co.uk" target="_blank">http://www.chicken-house.co.uk</a></td>
 </tr>
 <tr>
 <td>Silicone sheeting 0.3 mm translucent</td>
 <td></td>
 <td></td>
 <td><a href="http://www.silex.co.uk/" target="_blank">http://www.silex.co.uk/</a></td>
 </tr>
 <tr>
 <td>Image J software</td>
 <td></td>
 <td></td>
 <td><a href="http://rsbweb.nih.gov/ij/" target="_blank">http://rsbweb.nih.gov/ij/</a></td>
 </tr>
 <tr>
 <td>Zeiss OPMI pico</td>
 <td></td>
 <td></td>
 <td><a href="http://www.zeiss.co.uk/" target="_blank">http://www.zeiss.co.uk/</a></td>
 </tr>
 <tr>
 <td>Operating microscope</td>
 <td></td>
 <td></td>
 <td></td>
 </tr>
 <tr>
 <td>Vet tech solution isofluorane rig</td>
 <td></td>
 <td></td>
 <td><a href="http://www.vet-tech.co.uk/" target="_blank">http://www.vet-tech.co.uk/</a></td>
 </tr>
 <tr>
 <td>Vet tech solution isofluorane rig</td>
 <td></td>
 <td></td>
 <td><a href="http://www.vet-tech.co.uk/" target="_blank">http://www.vet-tech.co.uk/</a></td>
 </tr>
 </tbody>
</table>

in situ transverse rectus abdominis myocutaneous flap: a rat model of myocutaneous ischemia reperfusion injury

游离组织移植重建手术的金标准，不服从当地期权或那些需要的复合组织修复缺陷。缺血再灌注损伤（IRI）是一个已知的部分游离皮瓣失败的原因，有没有有效的治疗方法。建立这种损伤的实验室模型可以证明均付出沉重代价财政，大型哺乳动物通常用于在所需的专业知识，技术难度，这些程序通常需要聘请一位经验丰富的microsurgeon。本出版物和视频演示的典范IRI大鼠不需要显微专长的有效利用。此程序是一个原位模型的横向腹直肌肌皮瓣（TRAM）无创伤钳是用来重现缺血再灌注损伤与此手术相关。激光多普勒成像（LDI）扫描仪是用来评估皮瓣灌注和图像处理软件，重新评估面积百分比作为一个主要的测量结果受伤的皮肤生存的Image J。

比大动物模型大鼠模型更经济，36抗病性和基因操纵。松散的皮肤的动物，如啮齿类动物，被认为具有不同的安排相比，固定皮肤的动物如人类和猪的皮肤的血液供应。在宽松的皮肤的动物，皮肤直接皮肤血管，通过皮下脂肪相比之下覆皮肤（ 图4），主要提供固定剥皮的动物派生皮肤的血液供应，通过船只，当然通过底层肌肉提供覆包膜通过肌穿孔机（ 图4）。?...

Watch this Scientific Journal Video about In situ Transverse Rectus Abdominis Myocutaneous Flap: A Rat Model of Myocutaneous Ischemia Reperfusion Injury at JoVE.com

In situ Transverse Rectus Abdominis Myocutaneous Flap: A Rat Model of Myocutaneous Ischemia Reperfusion Injury

广泛采用游离组织移植重建手术，肿瘤切除术和创伤后恢复形式和功能。手术前预处理这个组织可能提高的结果。本文介绍了一种<em在原位</em&gt;横向腹直肌肌皮瓣（TRAM）大鼠作为测试预处理策略的一种手段。

在原位横向腹直肌肌皮瓣：肌缺血再灌注损伤模型大鼠

Free tissue transfer is the gold standard of reconstructive surgery to repair complex defects not amenable to local options or those requiring composite ...

in-situ-transverse-rectus-abdominis-myocutaneous-flap-rat-model

Research

JoVE Journal

Medicine

In situ Transverse Rectus Abdominis Myocutaneous Flap: A Rat Model of Myocutaneous Ischemia Reperfusion Injury

13.6K 次观看.  Royal Infirmary of Edinburgh. 广泛采用游离组织移植重建手术，肿瘤切除术和创伤后恢复形式和功能。手术前预处理这个组织可能提高的结果。本文介绍了一种

视频: In situ Transverse Rectus Abdominis Myocutaneous Flap: A Rat Model of Myocutaneous Ischemia Reperfusion Injury

Normothermic Cardiac Arrest and Cardiopulmonary Resuscitation: A Mouse Model of Ischemia-Reperfusion Injury

Acute Kidney Injury (AKI) is a common, highly lethal, complication of critical illness which has a high mortality1-4 and which is most 
frequently caused by whole-body hypoperfusion.5,6 Successful reproduction of whole-body hypoperfusion in rodent models has been fraught with 
difficulty.7-9,9,10 Models which employ focal ischemia have repeatedly demonstrated results which do not translate to the clinical setting, and larger 
animal models which allow for whole body hypoperfusion lack access to the full toolset of genetic manipulation possible in the mouse.11,12 However, in recent 
years a mouse model of cardiac arrest and cardiopulmonary resuscitation has emerged which can be adapted to model AKI.13 This model reliably reproduces 
physiologic, functional, anatomic, and histologic outcomes seen in clinical AKI, is rapidly repeatable, and offers all of the significant advantages of a murine surgical 
model, including access to genetic manipulative techniques, low cost relative to large animals, and ease of use. Our group has developed extensive experience 
with use of this model to assess a number of organ-specific outcomes in AKI.14,15

A powerful model for perioperative and critical care related acute kidney injury is presented. Using whole body hypoperfusion induced by cardiac arrest it is possible to nearly replicate the histologic and functional changes of clinical AKI.

常温心脏骤停和心肺复苏术：缺血再灌注损伤的小鼠模型

Acute Kidney Injury (AKI) is a common, highly lethal, complication of critical illness which has a high mortality1-4 and which is most 
frequently caused ...

科研

医学

Ischemia-reperfusion Model of Acute Kidney Injury and Post Injury Fibrosis in Mice

Ischemia-reperfusion induced acute kidney injury (IR-AKI) is widely used as a model of AKI in mice, but results are often quite variable with high, often unreported mortality rates that may confound analyses. Bilateral renal pedicle clamping is commonly used to induce IR-AKI, but differences between effective clamp pressures and/or renal responses to ischemia between kidneys often lead to more variable results. In addition, shorter clamp times are known to induce more variable tubular injury, and while mice undergoing bilateral injury with longer clamp times develop more consistent tubular injury, they often die within the first 3 days after injury due to severe renal insufficiency. To improve post-injury survival and obtain more consistent and predictable results, we have developed two models of unilateral ischemia-reperfusion injury followed by contralateral nephrectomy. Both surgeries are performed using a dorsal approach, reducing surgical stress resulting from ventral laparotomy, commonly used for mouse IR-AKI surgeries. For induction of moderate injury BALB/c mice undergo unilateral clamping of the renal pedicle for 26 min and also undergo simultaneous contralateral nephrectomy. Using this approach, 50-60% of mice develop moderate AKI 24 hr after injury but 90-100% of mice survive. To induce more severe AKI, BALB/c mice undergo renal pedicle clamping for 30 min followed by contralateral nephrectomy 8 days after injury. This allows functional assessment of renal recovery after injury with 90-100% survival. Early post-injury tubular damage as well as post injury fibrosis are highly consistent using this model.

We describe models of moderate and severe ischemia-reperfusion-induced kidney injury in which the mice undergo unilateral renal pedicle clamping followed by simultaneous or delayed contralateral nephrectomy, respectively. These models consistently give rise to renal dysfunction and post-injury fibrosis, but injury severity and survival are dependent on mouse background, age and surgical equipment.

急性肾损伤和伤后纤维化小鼠缺血再灌注模型

Ischemia-reperfusion induced acute kidney injury (IR-AKI) is widely used as a model of AKI in mice, but results are often quite variable with high, often ...

Induction and Assessment of Ischemia-reperfusion Injury in Langendorff-perfused Rat Hearts

The biochemical events surrounding ischemia reperfusion injury in the acute setting are of great importance to furthering novel treatment options for myocardial infarction and cardiac complications of thoracic surgery. The ability of certain drugs to precondition the myocardium against ischemia reperfusion injury has led to multiple clinical trials, with little success. The isolated heart model allows acute observation of the functional effects of ischemia reperfusion injury in real time, including the effects of various pharmacological interventions administered at any time-point before or within the ischemia-reperfusion injury window. Since brief periods of ischemia can precondition the heart against ischemic injury, in situ aortic cannulation is performed to allow for functional assessment of non-preconditioned myocardium. A saline filled balloon is placed into the left ventricle to allow for real-time measurement of pressure generation. Ischemic injury is simulated by the cessation of perfusion buffer flow, followed by reperfusion. The duration of both ischemia and reperfusion can be modulated to examine biochemical events at any given time-point. Although the Langendorff isolated heart model does not allow for the consideration of systemic events affecting ischemia and reperfusion, it is an excellent model for the examination of acute functional and biochemical events within the window of ischemia reperfusion injury as well as the effect of pharmacological intervention on cardiac pre- and postconditioning. The goal of this protocol is to demonstrate how to perform in situ aortic cannulation and heart excision followed by ischemia/reperfusion injury in the Langendorff model.

The isolated rat heart is an enduring model for ischemia reperfusion injury. Here, we describe the process of harvesting the beating heart from a rat via in situ aortic cannulation, Langendorff perfusion of the heart, simulated ischemia-reperfusion injury, and infarct staining to confirm the extent of ischemic insult.

诱导和缺血再灌注损伤评估中的Langendorff灌注鼠心

The biochemical events surrounding ischemia reperfusion injury in the acute setting are of great importance to furthering novel treatment options for ...

Murine Model of Intestinal Ischemia-reperfusion Injury

Intestinal ischemia is a life-threatening condition associated with a broad range of clinical conditions including atherosclerosis, thrombosis, hypotension, necrotizing enterocolitis, bowel transplantation, trauma and chronic inflammation. Intestinal ischemia-reperfusion (IR) injury is a consequence of acute mesenteric ischemia, caused by inadequate blood flow through the mesenteric vessels, resulting in intestinal damage. Reperfusion following ischemia can further exacerbate damage of the intestine. The mechanisms of IR injury are complex and poorly understood. Therefore, experimental small animal models are critical for understanding the pathophysiology of IR injury and the development of novel therapies.
Here we describe a mouse model of acute intestinal IR injury that provides reproducible injury of the small intestine without mortality. This is achieved by inducing ischemia in the region of the distal ileum by temporally occluding the peripheral and terminal collateral branches of the superior mesenteric artery for 60 min using microvascular clips. Reperfusion for 1 hr, or 2 hr after injury results in reproducible injury of the intestine examined by histological analysis. Proper position of the microvascular clips is critical for the procedure. Therefore the video clip provides a detailed visual step-by-step description of this technique. This model of intestinal IR injury can be utilized to study the cellular and molecular mechanisms of injury and regeneration.

Here we describe the detailed procedure of intestinal ischemia-reperfusion in mice which results in reproducible injury without mortality to encourage the standardization of this technique across the field. This model of intestinal ischemia-reperfusion injury can be utilized to study the cellular and molecular mechanisms of injury and regeneration.

肠缺血再灌注损伤的小鼠模型

Intestinal ischemia is a life-threatening condition associated with a broad range of clinical conditions including atherosclerosis, thrombosis, ...

A Mouse Model of Retinal Ischemia-Reperfusion Injury Through Elevation of Intraocular Pressure

Retinal ischemia-reperfusion (I/R) is a pathophysiological process contributing to cellular damage in multiple ocular conditions, including glaucoma, diabetic retinopathy, and retinal vascular occlusions. Rodent models of I/R injury are providing significant insights into mechanisms and treatment strategies for human I/R injury, especially with regard to neurodegenerative damage in the retinal neurovascular unit. Presented here is a protocol for inducing retinal I/R injury in mice through elevation of intraocular pressure (IOP). In this protocol, the ocular anterior chamber is cannulated with a needle, through which flows the drip of an elevated saline reservoir. Using this drip to raise IOP above systolic arterial blood pressure, a practitioner temporarily halts inner retinal blood flow (ischemia). When circulation is reinstated (reperfusion) by removal of the cannula, severe cellular damage ensues, resulting ultimately in retinal neurodegeneration. Recent studies demonstrate inflammation, vascular permeability, and capillary degeneration as additional elements of this model. Compared to alternative retinal I/R methodologies, such as retinal arterial ligation, retinal I/R injury by elevated IOP offers advantages in its&#160;anatomical specificity, experimental tractability, and technical accessibility, presenting itself as a valuable tool for examining neuronal pathogenesis and therapy in the retinal neurovascular unit.

This article describes a procedure for inducing retinal ischemia-reperfusion injury by elevated intraocular pressure in mice. Retinal ischemia-reperfusion injury by elevated intraocular pressure serves to model human pathologies characterized by compromised oxygen and nutrient delivery in the retina, enabling researchers to examine potential cellular mechanisms and treatments for human diseases of the retinal neurovascular unit.

视网膜缺血再灌注损伤的小鼠模型通过眼压升高

Retinal ischemia-reperfusion (I/R) is a pathophysiological process contributing to cellular damage in multiple ocular conditions, including glaucoma, ...

Evaluation of Coronary Flow Reserve After Myocardial Ischemia Reperfusion in Rats

Coronary artery disease is the leading cause of death worldwide. After an acute myocardial infarction, early and successful myocardial intervention via recanalization of the coronary artery is the most effective strategy for reducing the size of ischemic myocardium. The coronary microvasculature cannot be visualized and imaged&#160;in vivo, but there are several invasive and noninvasive techniques that can be used to assess parameters which depend directly on coronary microvascular function. The endothelial function after ischemia reperfusion can be assessed also at the level of the coronary circulation via the coronary flow reserve (CFR).&#160;In this study, peak velocity of left anterior descending (LAD) coronary arteries was measured in rats in vivo via Transthoracic Doppler Echocardiography during resting and stress challenge (induced by Dobutamine). A normal heart can increase its coronary blood flow up to four times above the resting values during stress induction. Following ischemia reperfusion, we found a significantly diminished CFR, which can be used as a marker of coronary microvascular dysfunction. CFR has opened a window on the importance of microvascular dysfunction and has been shown to predict cardiovascular risk independent of whether the severe obstructive disease is present.

Coronary flow reserve (CFR), is defined as the ratio of maximal coronary blood flow to the resting coronary blood flow. We present a protocol for evaluating CFR in rats via ultrasound, which offers the opportunity to predict cardiovascular risk factors in the absence of obstructive coronary disease.

大鼠心肌缺血再灌注后冠状动脉流动储备评价

Coronary artery disease is the leading cause of death worldwide. After an acute myocardial infarction, early and successful myocardial intervention via ...

Modified Heterotopic Hindlimb Osteomyocutaneous Flap Model in the Rat for Translational Vascularized Composite Allotransplantation Research

Vascularized composite allotransplantation (VCA) is a relatively new field in the reconstructive surgery. Clinical achievements in human VCA include hand and face transplants and, more recently, abdominal wall, uterus, and urogenital transplants. Functional outcomes have exceeded initial expectations, and most recipients enjoy an improved quality of life. However, as clinical experience accumulates, chronic rejection and complications from the immunosuppression must be addressed. In many cases where grafts have failed, the causative pathology has been ischemic vasculopathy. <span style="font-size: 11pt; line-height: 115%; font-family: Arial, sans-serif; color: rgb(41, 43, 49); background-image: initial; background-position: initial; background-size: initial; background-repeat: initial; background-attachment: initial; background-origin: initial; background-clip: initial;">The biological mechanisms of the acute and chronic rejection associated with VCA, especially ischemic vasculopathy, are important areas of research. However, due to the very small number of VCA patients, the evaluation of proposed mechanisms is better addressed in an experimental model. Multiple groups have used animal models to address some of the relevant unsolved questions in VCA rejection and vasculopathy. Several model designs involving a variety of species are described in the literature. Here we present a reproducible model of VCA heterotopic hindlimb osteomyocutaneous flap in the rat that can be utilized for translational VCA research. This model allows for the serial evaluation of the graft, including biopsies and different imaging modalities, while maintaining a low level of morbidity.

Vascularized composite allograft offers life-altering benefits to transplant recipients, but the biological causes of graft rejection and vasculopathy remain poorly understood. The rodent surgical model presented here offers a reproducible, clinically relevant model of transplantation, allowing researchers to evaluate rejection events and potential therapeutic strategies to prevent their occurrence.

改良异位异体下肢骨皮瓣模型在大鼠转化血管化复合同种异体移植研究中的应用

Vascularized composite allotransplantation (VCA) is a relatively new field in the reconstructive surgery. Clinical achievements in human VCA include hand ...

A Pre-clinical Rat Model for the Study of Ischemia-reperfusion Injury in Reconstructive Microsurgery

Ischemia-reperfusion injury is the main cause of flap failure in reconstructive microsurgery. The rat is the preferred preclinical animal model in many areas of biomedical research due to its cost-effectiveness and its translation to humans. This protocol describes a method to create a preclinical free skin flap model in rats with ischemia-reperfusion injury. The described 3 cm x 6 cm rat free skin flap model is easily obtained after the placement of several vascular ligatures and the section of the vascular pedicle. Then, 8 h after the ischemic insult and completion of the microsurgical anastomosis, the free skin flap develops the tissue damage. These ischemia-reperfusion injury-related damages can be studied in this model, making it a suitable model for evaluating therapeutic agents to address this pathophysiological process. Furthermore, two main monitoring techniques are described in the protocol for the assessment of this animal model: transit-time ultrasound technology and laser speckle contrast analysis.

Here, we describe a pre-clinical animal model for studying the pathophysiology of ischemia-reperfusion injury in reconstructive microsurgery. This free skin flap model based on the superficial caudal epigastric vessels in the rat may also allow for the evaluation of different therapies and compounds to counteract ischemia-reperfusion injury-related damage.

重建显微外科缺血再灌注损伤研究的临床前大鼠模型

Ischemia-reperfusion injury is the main cause of flap failure in reconstructive microsurgery. The rat is the preferred preclinical animal model in many ...

Delayed Intramyocardial Delivery of Stem Cells after Ischemia Reperfusion Injury in a Murine Model

There is significant interest in the use of stem cells (SCs) for the recovery of cardiac function in individuals with myocardial injuries. Most commonly, cardiac stem cell therapy is studied by delivering SCs concurrently with the induction of myocardial injury. However, this approach presents two significant limitations: the early hostile pro-inflammatory ischemic environment may affect the survival of transplanted SCs, and it does not represent the subacute infarction scenario where SCs will likely be used. Here we describe a two-part series of surgical procedures for the induction of ischemia-reperfusion injury and delivery of mesenchymal stem cells (MSCs). This method of stem cell administration may allow for the longer viability and retention around damaged tissue by circumventing the initial immune response. A model of ischemia reperfusion injury was induced in mice accompanied by the delivery of mesenchymal stem cells (3.0 x 105), stably expressing the reporter gene firefly luciferase under the constitutively expressed CMV promoter, intramyocardially 7 days later. The animals were imaged via ultrasound and bioluminescent imaging for confirmation of injury and injection of cells, respectively. Importantly, there was no added complication rate when performing this two-procedure approach for SC delivery. This method of stem cell administration, collectively with the utilization of state-of-the-art reporter genes, may allow for the in vivo study of viability and retention of transplanted SCs in a situation of chronic ischemia commonly seen clinically, while also circumventing the initial pro-inflammatory response. In summary, we established a protocol for the delayed delivery of stem cells into the myocardium, which can be used as a potential new approach in promoting regeneration of the damaged tissue.

Stems cells are continuously investigated as potential treatments for individuals with myocardial damage, however, their decreased viability and retention within injured tissue can impact their long-term efficacy. In this manuscript we describe an alternative method for stem cell delivery in a murine model of ischemia reperfusion injury.

在穆林模型中缺血再灌注损伤后干细胞的心肌内传递延迟

There is significant interest in the use of stem cells (SCs) for the recovery of cardiac function in individuals with myocardial injuries. Most commonly, ...

Organ Ischemia-Reperfusion Injury by Simulating Hemodynamic Changes in Rat Liver Transplant Model

Orthotopic liver transplantation (OLT) in rats is a tried and proven animal model used for preoperative, intraoperative, and postoperative studies, including ischemia-reperfusion injury (IRI) of extrahepatic organs. This model requires numerous experiments and devices. The duration of anhepatic phase is closely related to the time to develop IRI after transplantation. In this experiment, we used hemodynamic changes to induce extrahepatic organ damage in rats and determined the maximum tolerance time. The time until the most severe organ injury varied for different organs. This method can easily be replicated and can also be used to study IRI of the extrahepatic organs after liver transplantation.

This paper provides a detailed description of how to build an animal model of the anhepatic phase (liver ischemia) in rats to facilitate basic research into ischemia-reperfusion injury after liver transplantation.

器官缺血-输液损伤模拟大鼠肝脏移植模型的血动力学变化

Orthotopic liver transplantation (OLT) in rats is a tried and proven animal model used for preoperative, intraoperative, and postoperative studies, ...