The authors would like to acknowledge funding support from the University of South Florida, Department of Surgery (Mack Wu, MD, PhD).

来自南佛罗里达州的动物护理委员会（IACUC）的大学，获得批准​​下文提出的所有动物的程序，并通过动物福利法和指南护理和使用实验动物的所有要求遵守:注意。 1.准备硅床单和手术器械<ol><li>预切割的条带（10.5厘米×3.0cm）的非增强0.01厚度，医用级硅酮薄片和消毒使用高压釜。 </li><li>清洁和消毒适合手术器械（剪刀，镊子和窗帘或毛巾在手术过程中创造一个无菌区）。 </li></ol> 2.实验动物<ol><li>使用成年男性或女性老鼠从商业饲养员获得250-350克范围内称量。如果老年大鼠被利用，它们应≥350克，以确保更好地存活手术后。在此之前的任何实验，开始适应所有的动物，至少一个12小时的光-暗循环的食物和水随意的标准条件下7天。 </li></ol> 3.麻醉，手术前镇痛和手术准备<ol><li>通过经由一个感应腔室使用异氟烷在3％-4％诱导全身麻醉和（经由使用鼻锥）在1％维持-2％与皮肤制剂和手术过程中的O 2。由观察的速率和呼吸，叉指捏或眼睑眨眼反射深度监测麻醉深度。 注意:这时，兽医软膏可以放在眼睛，以防止干燥时，动物是在麻醉状态下。 </li><li>从无菌手术区的位置偏远，将大鼠俯卧位，并从颈部向下约11厘米的基础剃胡须快船背部。钢网，记号笔，轮廓为3.0厘米×10.5厘米瓣（ 见图1A）。 </li><li>移动老鼠到一个干净的，指定surgical区域配有经批准的加热垫和无菌手术单或毛巾。注射5毫克/公斤皮下酮洛芬之前第一手术切口疼痛管理。附加的流体（盐水）可给予（高达5毫升）皮下需要。 </li><li>进一步准备皮肤通过擦拭先用70％异丙醇和第二用0.2％氯己定，然后应用灭菌窗帘来创建无菌区。 10％的聚维酮碘（聚维酮碘）也可以使用。 注:抗生素（氨苄青霉素，15毫克/千克）可经皮下施用，但如果使用好的无菌技术不是必需的。 </li></ol> 4.创建切除的创伤和Bipedicled皮瓣<ol><li>使用无菌的，一次性6毫米活检穿孔工具，在指定的皮瓣区域（ 图1B）的中心创建两个圆形"缺血"伤口。伤口的深度应该是向下（不通过）的肉膜carno底层筋膜SUS肌肉（ 图1B插图）。 </li><li>使用镊子解除在由冲床活检产生的伤口轮廓的中间的皮肤，然后使用虹膜剪（带有弯曲提示）切除圆形片组织（包括肉膜肌）。其结果将是一个全厚伤口与筋膜作为伤口的底部。 注:切除的组织（伤口塞）可以被快速冷冻在液氮中，或固定在10％缓冲的甲醛O / N购买处理作为对照，正常的皮肤。 </li><li>创建通过使切口与在缺血性伤口沿预先绘制的线（ 图1C）是10.5厘米长和3.0厘米外的每一侧的无菌解剖刀一个bipedicled瓣。切口的深度应下降到椎旁肌肉。使用虹膜剪刀，从椎旁肌分开肌层筋膜，小心翼翼地保持完好筋膜作为"基地"6毫米拳（Figure 1D）。 </li><li>取1无菌预切硅氧烷片，并将其放置在肉膜筋膜和脊柱旁的肌肉（ 图1E），确保纸张不会弯曲或折叠之间。使用黑色，非吸收性缝线（大小4.0）由锚定硅酮片给皮肤的至少8中断每侧线圈关闭两个切口，沿着翼片（ 图1F和1G）的长度。 </li><li>使用无菌的，一次性活检穿孔工具，创建两个内部控制"非缺血"伤口（下到肉膜肌肉的前筋膜）1厘米横向于缺血性伤口上的折翼（ 图1G）的任一侧。 </li><li>将下面的伤口一把尺子和拍摄数码照片伤口测量目的（参见图3A）。此时，血流量（灌注）可以通过使用激光多普勒或执行其他操作（外用药物放置）的监测。</li><li>适用于经批准的液体胶粘剂无论头部和尾部的伤口和透明薄膜敷料，保持伤口湿润的环境和清洁（无菌）。一个附加的敷料可以被放置在护翼的尾端，以防止动物从除去最尾的缝合线。 </li></ol> 5.术后程序<ol><li>放置动物在笼中（单独地收容）配备有浅馈线，以防止外科手术部位从摩擦馈线。动物应该有人看管或返回给其他动物的公司，直到他们重新获得足够的意识，保持胸骨斜卧并表现出有目的的运动。在恢复过程中的加热垫应置于半笼长达2天。 </li><li>为了术后疼痛管理，管理酮洛芬（5毫克/千克）皮下动物第二天早上和每天1个长达48小时后手术。动物也应每天监测疼痛，体重减轻或手术部位感染的延长迹象。 </li></ol> 6.随后伤口测量和换药<ol><li>通过一个感应室下使用异氟烷麻醉频繁测量缺血性和非缺血性伤口在3％-4％，并在1％-2％与O 2的保持（经由鼻锥）如在步骤3.1。 </li><li>轻轻除去敷料，以不拉从皮肤粘合。这时附加的数字照片都是伤口测量，局部治疗重施，激光多普勒成像（LDI），或执行，以满足研究者的需要的其他操作。 </li><li>适用的粘合剂和干净的敷料，并允许动物恢复如在步骤5.1。 </li></ol> 7.伤口收集和安乐死<ol><li>收获缺血性和非缺血性创伤（天研究者认​​为适当），而动物是在全身麻醉下，通过一个IND使用异氟烷在3％-4％，减税室，并在1％-2％与O 2的保持（经由鼻锥）如在步骤3.1。 </li><li>使用手术刀，使伤口周围的正方形形状的切除，包括伤口周围的一些健康组织。将切除成1.5毫升卡扣帽管和捕捉​​冷冻在液氮（存储于-80℃）为未来的分子分析或孵育，在10％缓冲的福尔马林中O / N在RT组织学处理。 注:伤口切除，也可以减少一半，以提供更多的样本进行分析。 </li><li>伤口组织切除后，用二氧化碳吸入法批准安乐死的动物。 </li></ol>

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The authors have nothing to disclose.

<html>大鼠伤口愈合经常被讨论的主题，因为它们愈合伤口感染和动物间差异5率高的能力。它的一个发展过程中的模型的最初目标是要减少这种变化。修改到护翼的宽度，减少伤口放置特定的数量（集中在具有一致的头尾位置的翼片）的硅酮片材的和引入已经完成了这个目标。通过收缩伤口愈合也已减少和治疗上皮，如在人类中，是测得的结果。该模型以不同的菌株鼠适应， 即 F344，也证明是成功的，并再现缺血程度采用SD大鼠观察。整体所需此模型中的外科技术（活检，缝合和伤口切除）容易被大多数学生和技术人员有限手术经验获得的。 _content"&gt;要实现与该模型的一致性，同时执行多次手术，我们发现它创建之前皮瓣硅氧烷纸张安置5的仰角的缺血性伤口是重要的。另外，通过肉膜筋膜不冲孔是关键提供了可行的伤口床保持在硅氧烷。硅氧烷不仅作用，以防止血管再生长，但也可作为一个"夹板"，鼓励伤口再上皮。粘合剂和敷料中的应用，以防止感染并保持湿润的环境伤口愈合也很重要。产​​品的选择可以是什么，优选或在研究者的动物设施中使用，然而，它并不少见一些动物，以便能够除去其敷料，不管粘合剂的什么类型/敷料的组合是使用。 该bipedicled瓣应在整个治疗的时间进程是近似保持活力<html> LY 28天，这取决于大鼠品系和其它共病存在。很少，脓肿可在皮瓣（特别是靠近缝线）和血清肿形成可能形成瓣下。流体可以被排出和抗生素给药，如果必要的。然而，如果襟翼失去生存能力和变得坏死，建议该动物不再被使用。伤口切除用于生化分析确实引入变异由于（1）一些正常组织必须保留支撑件（2）的组织均质化，制备的选择的RNA，DNA或蛋白质，（3）固有动物间差异5,10隔离， 11。人们可以考虑这最后一点限制到模型并发现降低了皮瓣（&lt;2.0厘米）或襟翼创伤的大小可引起坏死，这表明在因素，如温度或压力水平的微小变化，也可能导致到伤口样本之间生化检测变化从一个老鼠另外5。 ve_content"&gt;总之，本模型中，具有纵向，bipedicled瓣范围从2.0-3.0厘米的宽度和策略性地放置硅酮片，是长时间的组织局部缺血的一个可靠的模型。一旦用户是擅长使用的技术，以创建一致的缺血性创伤，它们应能使其适应啮齿动物的额外的年龄和物种（包括小鼠）的切除伤口可以局部治疗，或用于进一步探索涉及慢性伤口的形成机制（多个）全身治疗，加剧了炎症反应，异常血管生成和延迟愈合。

有效的药物开发等伤口愈合的治疗需要适当的体内模型，尽管翻译在动物模型的研究结果为人类治疗1已知问题。以下是对使用局部缺血皮肤伤口愈合的大鼠模型的详细协议的描述，调查机制的病理伤口愈合进一步的理解。鼠种，经常采用，由于其广泛的可用性，尺寸和用于驯服自然伤口愈合研究，因为它是足够大以提供一个合适的皮肤面积为切口和切除伤人，成像和组织收集2。然而，应当考虑到仔细考虑了大鼠和人的皮肤上是不同的解剖学上，用大鼠被称为松散皮肤动物。这显着特点，使得伤口收缩，而不是上皮到显著到大鼠皮肤闭幕瓦特贡献ounds 2。此外，皮下肉膜肌的大鼠的存在下，通过双方的收缩和胶原形成3,4-有助于愈合。这些非常重要的解剖区分被认为是在大鼠缺血性皮肤伤口模型的发展和具体的修改被实施，以减少伤口收缩，减少肉膜肌5的影响。 在糖尿病足部溃疡，下肢静脉溃疡和压力性溃疡，愈合被延迟和这些伤口被认为是慢性的。伤口的特征在于过度的炎症，其防止从伤口进展到伤口修复6的下一个阶段。一种在慢性伤口的发展的主要因素是局部组织缺血（血流量减少）5有助于无法清除炎症。在这期间这款机型正在开发和验证（2003-2004年中的时候），没有标准化的动物模型，可以提供足够的组织以测试诱导血管生成的伤口床，在正常的伤口愈合的关键阶段和动机发展此模型5。这就是说，这里提出的模型是最初由Schwartz 等7和随后描述的缺血性伤口模型的变形由Chen 等人所用的修饰的形式。8 在修改后的缺血性伤口模型，已经作了修改，以规避的大鼠的上述解剖特征，导致治疗效果的收缩，而不是上皮:（1）两个全层切除伤口内bipedicled背部皮肤挡板和创建肉膜肌从由解剖正上方的肌肉筋膜床伤口移除。 （2）在翼片本身具有位于在t的中点更窄的尺寸，以确保血液供应是随机的，伤口他皮瓣缺血是。 （3）一种硅片被插入挡板下面，以防止readherence从下层组织瓣的再灌注。伤口收缩是有限的（不排除）由固定或缝合皮瓣硅胶片5。 该模型最近已应用在研究范围从对缺血性伤口愈合9,10-缺血伤口愈合高压氧影响在年轻与老年大鼠11，并已被证明是长期组织缺血的一个可靠的模型。在bipedicled瓣的尺寸也已经适应了不同的大鼠品系，包括只SD（11厘米长2厘米宽）和F344大鼠（10.5厘米长按3.0-3.5厘米宽）和其他物种，包括猪12鼠13,14。这部影片采用了缺血性皮肤创伤模型的演示F344近交系大鼠品系。 审批下面提出的所有动物的程序是获得编辑从南佛罗里达州的动物护理委员会（IACUC）的大学和动物福利法案和指南护理和使用实验动物的所有要求遵守。

<table><tbody><tr><td>Sil-Tec medical grade sheeting</td><td>Technical Products Inc.</td><td>500-3</td><td>nonreinforced, 0.01 inches in thickness</td></tr><tr><td>Mini Iris scissors, 8 cm, curved, SS</td><td>World Precision Instruments</td><td>#503671</td><td></td></tr><tr><td>Ethilon Nylon Sutures</td><td>Ethicon</td><td>1964G</td><td>black, size 4.0, PC-3 16&amp;nbsp;mm needles (3/8 circle)</td></tr><tr><td>Laser Doppler Imager</td><td>Moor Instruments</td><td>moorLDI2-IR</td><td>Standard blood flow imager: http://us.moor.co.uk/product/moorldi2-laser-doppler-imager/8</td></tr><tr><td>ImageJ</td><td>NIH</td><td>free download</td><td>http://rsb.info.nih.gov/ij/</td></tr><tr><td>Mastisol</td><td>Henry Schein</td><td>Cat # 7289210</td><td>Fisher Scientific NC9774929</td></tr><tr><td>Tegaderm</td><td>Medical Specialties&amp;nbsp;</td><td>3M1624W</td><td>Fisher Scientific NC9922128</td></tr></tbody></table>

demonstration of the rat ischemic skin wound model

在人类的倾向慢性伤口增加了与老化，疾病状况如糖尿病和心血管障碍的功能，并且由于动减轻的压力。动物模型已经开发了试图模仿这些条件促进我们的慢性伤口的复杂性的理解的目的。本文所描述的模型是大鼠缺血皮瓣模型允许长期减少导致伤口成为局部缺血和类似慢性伤口的表型（缩血管化，增加了炎症和延迟伤口闭合）血流。它由一个bipedicled背瓣集中放置2缺血性伤口和2个非缺血性伤口横向到瓣作为对照。一种新型除此之外缺血皮瓣模型是安置在充当屏障，夹板，以防止血管再通，减少收缩的伤口愈合瓣下方的硅胶片的。尽管使用大鼠伤口愈合研究，由于其相比于人类相当不同解剖和生理差异的争论（ 即一个肉膜肌，短寿命的存在下，毛囊的数量增加，并且其愈合伤口感染能力）在这个模型中采用的修改，使其成为一个有价值的替代以前开发的缺血皮瓣模型。

大鼠缺血性伤口愈合模型协议应该采取每头约20分钟，如果有效地执行。此前申请的敷料的模型应该出现如图1G表示。这将是重要的，以验证该bipedicled襟翼和伤口在其中是缺血。皮下氧张力（PSCO 2）在伤口的水平这一模型5的验证期间被测量通过放置一个极谱电极在两个缺血性伤口之间的皮下组织。 PSCO 2值分别在危重缺血范围（20-40毫米汞柱）。由于这种模式的发展运用LDI已越来越多地被用于测量血液灌注，这种技术将提供有关bipedicled皮瓣缺血状态的足够信息。 简言之，LDI技术是基于由一个光纤探针进行激光光束的发射。该measuri纳克深度取决于组织性质的激光和波长。在正常的皮肤，与标准纤维分离（0.25毫米）和780纳米波长的激光探针的仪器，测量深度将是0.5-1毫米的量级。代表多普勒图像（左），用于背侧bipedicled襟翼同时显示一个灌注区域（缝合线的左侧）和非灌注区域（缝合线的右边）示于图2中 。 除了测量皮下氧张力，还可以使用探针或共同的生化标志物，以建立该伤口皮瓣是缺血。 PECAM-1或CD31上正常的血管内皮细胞上发现是一个标记为新血管形成中的创伤。各种标记物的活性氧，发现在缺血性伤口中升高，是市售的常作为荧光标记的抗体或超指标如二氢乙（DHE）。 Wounð面积测量来追踪伤口闭合可以以各种方式来表示。通常，卷绕区域从伤口愈合超过11的时间过程使用公式的数字图像，例如量化（6毫米冲床活检面积=（π）R 2 = 3.14×9 =28.26毫米2在第0天）的数据表现为初始伤口面积或百分比在某一天创面面积可以量化为在古尔德等人 5出于演示的目的，伤口愈合的进展表示为％的初始伤口面积的28天时间过程。使用自由软件ImageJ的，数字图像被打开和规模，用10毫米的图像（ 图3A）在标尺上设置。在长度拉伸10mm的线相当于一个象素计数（ 中的插入图3A），它可以被转换成所选择的一个单元，在这种情况下（毫米）。接着，对伤口的圆周被追踪在图像上（图3B），并且一旦测量命令被给出，该区域被显示在毫米（中的插入图3B）。然后数据可被呈现为在y轴和天，x轴（ 图3C）初始伤口面积的百分数。 <img alt="图1" src="/files/ftp_upload/52637/52637fig1.jpg" /> 图1的照片描绘了在手术过程中的步骤来创建缺血性伤口时，bipedicled瓣，和非缺血性伤口。 （A）术前脱毛和皮肤准备麻醉的老鼠接受的镇痛药（酮洛芬）皮下注射术前剂量的疼痛管理。（B和插图）缺血性伤口里面的2切口标记创建利用无菌冲活检工具。（C）的切口沿着标线所作下降到脊柱旁肌肉和（D）的bipedicled襟翼被升高显示瓣的分离（用肉膜筋膜完好）从下面的肌肉层。（E）的无菌硅氧烷（黑色箭头）被放置在肌层的筋膜和脊柱旁的肌肉之间的片（F）第黑，非吸收性缝线（大小4.0）由锚定硅酮片给皮肤与多个中断线圈沿着翼片（G）的长度关闭这两个切口。两个非缺血性伤口（黑色箭头）使用无菌穿刺活检工具横向到双方的bipedicled瓣创建。 <a href="https://www.jove.com/files/ftp_upload/52637/52637fig1large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <img alt="图2" src="/files/ftp_upload/52637/52637fig2.jpg" /> 血液灌流手术后的图2代表性的激光多普勒图像，右平移埃尔显示2-缺血性伤口（黑色箭头）在bipedicled瓣的中间和1个非缺血性伤口（单一的黑色箭头）横向于挡板的黑白图像。缝合线也以白色被强调。左边的面板显示在右侧面板中描绘的同一区域的多普勒图像。用鲜艳的颜色方面不是与深蓝色的区域更灌注。在非缺血区（左）和局部缺血区（右）之间灌注这种差异是清楚的，并且可以随后沿着缝合线的整个长度。需要注意的是双方的伤口显得明亮，如血细胞仍然存在，在一定程度上。 <a href="https://www.jove.com/files/ftp_upload/52637/52637fig2large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <img alt="图3" src="/files/ftp_upload/52637/52637fig3.jpg" /> 图3.伤口的测量和数据表示。 </st荣&gt;（A）中的数字照片，描绘用软件（ImageJ的）伤口面积的测量，转换像素以（毫米）（黑色箭头），以及方法来捕获创伤（B）的一个准确的圆周（单一的黑色箭头）。统计数据（平均伤口面积的±SEM在某一天）然后可以被呈现为初始伤口面积（第0天）在y轴和天，x轴（C）的百分比。使用双因子ANOVA用Sidak多重比较试验的曲线图呈现的数据进行分析，***表示显著高（P = 0.0004）比非缺血性在相同时间点（10天）中，N = 8的伤口每组。 <a href="https://www.jove.com/files/ftp_upload/52637/52637fig3large.jpg" target="_blank">请点击此处查看该图的放大版本。</a>

Watch this Scientific Journal Video about Demonstration of the Rat Ischemic Skin Wound Model at JoVE.com

Demonstration of the Rat Ischemic Skin Wound Model

鼠，由于它的尺寸，可用性，和相当温顺的行为，已经被用作研究模型多年。该协议的目的是利用老鼠作为缺血性皮肤伤口愈合模型提供有价值的洞察慢性伤口的病理生理学。

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 ...

demonstration-of-the-rat-ischemic-skin-wound-model

Nanyang Technological University

Research

JoVE Journal

Medicine

21.3K 次观看.  University of South Florida. 鼠，由于它的尺寸，可用性，和相当温顺的行为，已经被用作研究模型多年。该协议的目的是利用老鼠作为缺血性皮肤伤口愈合模型提供有价值的洞察慢性伤口的病理生理学。 

视频: Demonstration of the Rat Ischemic Skin Wound Model

Acute Myocardial Infarction in Rats

With heart failure leading the cause of death in the USA (Hunt), biomedical research is fundamental to advance medical treatments for cardiovascular diseases. Animal models that mimic human cardiac disease, such as myocardial infarction (MI) and ischemia-reperfusion (IR) that induces heart failure as well as pressure-overload (transverse aortic constriction) that induces cardiac hypertrophy and heart failure (Goldman and Tarnavski), are useful models to study cardiovascular disease. In particular, myocardial ischemia (MI) is a leading cause for cardiovascular morbidity and mortality despite controlling certain risk factors such as arteriosclerosis and treatments via surgical intervention (Thygesen). Furthermore, an acute loss of the myocardium following myocardial ischemia (MI) results in increased loading conditions that induces ventricular remodeling of the infarcted border zone and the remote non-infarcted myocardium. Myocyte apoptosis, necrosis and the resultant increased hemodynamic load activate multiple biochemical intracellular signaling that initiates LV dilatation, hypertrophy, ventricular shape distortion, and collagen scar formation. This pathological remodeling and failure to normalize the increased wall stresses results in progressive dilatation, recruitment of the border zone myocardium into the scar, and eventually deterioration in myocardial contractile function (i.e. heart failure). The progression of LV dysfunction and heart failure in rats is similar to that observed in patients who sustain a large myocardial infarction, survive and subsequently develops heart failure (Goldman). The acute myocardial infarction (AMI) model in rats has been used to mimic human cardiovascular disease; specifically used to study cardiac signaling mechanisms associated with heart failure as well as to assess the contribution of therapeutic strategies for the treatment of heart failure. The method described in this report is the rat model of acute myocardial infarction (AMI). This model is also referred to as an acute ischemic cardiomyopathy or ischemia followed by reperfusion (IR); which is induced by an acute 30-minute period of ischemia by ligation of the left anterior descending artery (LAD) followed by reperfusion of the tissue by releasing the LAD ligation (Vasilyev and McConnell). This protocol will focus on assessment of the infarct size and the area-at-risk (AAR) by Evan's blue dye and triphenyl tetrazolium chloride (TTC) following 4-hours of reperfusion; additional comments toward the evaluation of cardiac function and remodeling by modifying the duration of reperfusion, is also presented. Overall, this AMI rat animal model is useful for studying the consequence of a myocardial infarction on cardiac pathophysiological and physiological function.

The rat model of acute myocardial infarction (AMI) is useful to study the consequence of a MI on cardiac pathophysiological and physiological function.

大鼠急性心肌梗死

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

科研

医学

A Simplified Technique for Producing an Ischemic Wound Model

One major obstacle in current diabetic wound research is a lack of an ischemic wound model that can be safely used in diabetic animals. Drugs that work well in non-ischemic wounds may not work in human diabetic wounds because vasculopathy is one major factor that hinders healing of these wounds. We published an article in 2007 describing a rabbit ear ischemic wound model created by a minimally invasive surgical technique. Since then, we have further simplified the procedure for easier operation. On one ear, three small skin incisions were made on the vascular pedicles, 1-2 cm from the ear base. The central artery was ligated and cut along with the nerve. The whole cranial bundle was cut and ligated, leaving only the caudal branch intact. A circumferential subcutaneous tunnel was made through the incisions, to cut subcutaneous tissues, muscles, nerves, and small vessels. The other ear was used as a non-ischemic control. Four wounds were made on the ventral side of each ear. This technique produces 4 ischemic wounds and 4 non-ischemic wounds in one animal for paired comparisons. After surgery, the ischemic ear was cool and cyanotic, and showed reduced movement and a lack of pulse in the ear artery. Skin temperature of the ischemic ear was 1-10 &deg;C lower than that on the normal ear and this difference was maintained for more than one month. Ear tissue high-energy phosphate contents were lower in the ischemic ear than the control ear. Wound healing times were longer in the ischemic ear than in the non-ischemic ear when the same treatment was used. The technique has now been used on more than 80 rabbits in which 23 were diabetic (diabetes time ranging from 2 weeks to 2 years). No single rabbit has developed any surgical complications such as bleeding, infection, or rupture in the skin incisions. The model has many advantages, such as little skin disruption, longer ischemic time, and higher success rate, when compared to many other models. It can be safely used in animals with reduced resistance, and can also be modified to meet different testing requirements.

We have developed a minimally invasive technique to create a rabbit ischemic ear wound model by dividing the central artery and nerve and the cranial neurovascular bundle. A subcutaneous tunnel then cuts all subcutaneous tissues. This procedure causes minimal skin disruption and can be safely used in diabetic animals.

一个生产缺血性损伤模型的简化技术

One major obstacle in current diabetic wound research is a lack of an ischemic wound model that can be safely used in diabetic animals. Drugs that work ...

Murine Model of Wound Healing

Wound healing and repair are the most complex biological processes that occur in human life. After injury, multiple biological pathways become activated. Impaired wound healing, which occurs in diabetic patients for example, can lead to severe unfavorable outcomes such as amputation. There is, therefore, an increasing impetus to develop novel agents that promote wound repair. The testing of these has been limited to large animal models such as swine, which are often impractical. Mice represent the ideal preclinical model, as they are economical and amenable to genetic manipulation, which allows for mechanistic investigation. However, wound healing in a mouse is fundamentally different to that of humans as it primarily occurs via contraction. Our murine model overcomes this by incorporating a splint around the wound. By splinting the wound, the repair process is then dependent on epithelialization, cellular proliferation and angiogenesis, which closely mirror the biological processes of human wound healing. Whilst requiring consistency and care, this murine model does not involve complicated surgical techniques and allows for the robust testing of promising agents that may, for example, promote angiogenesis or inhibit inflammation. Furthermore, each mouse acts as its own control as two wounds are prepared, enabling the application of both the test compound and the vehicle control on the same animal. In conclusion, we demonstrate a practical, easy-to-learn, and robust model of wound healing, which is comparable to that of humans.

A murine model of cutaneous wound healing that can be used to assess therapeutic compounds in physiological and pathophysiological settings.

伤口愈合的小鼠模型

Wound healing and repair are the most complex biological processes that occur in human life. After injury, multiple biological pathways become activated. ...

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

Free tissue transfer is the gold standard of reconstructive surgery to repair complex defects not amenable to local options or those requiring composite tissue. Ischemia reperfusion injury (IRI) is a known cause of partial free flap failure and has no effective treatment. Establishing a laboratory model of this injury can prove costly both financially as larger mammals are conventionally used and in the expertise required by the technical difficulty of these procedures typically requires employing an experienced microsurgeon. This publication and video demonstrate the effective use of a model of IRI in rats which does not require microsurgical expertise. This procedure is an in situ model of a transverse abdominis myocutaneous (TRAM) flap where atraumatic clamps are utilized to reproduce the ischemia-reperfusion injury associated with this surgery. A laser Doppler Imaging (LDI) scanner is employed to assess flap perfusion and the image processing software, Image J to assess percentage area skin survival as a primary outcome measure of injury.

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

Free tissue transfer is widely employed in reconstructive surgery to restore form and function following oncological resection and trauma. Preconditioning this tissue prior to surgery may improve outcome. This article describes an in situ transverse rectus abdominis myocutaneous flap (TRAM) in rats as a means for testing preconditioning strategies.

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 ...

Ischemic Tissue Injury in the Dorsal Skinfold Chamber of the Mouse: A Skin Flap Model to Investigate Acute Persistent Ischemia

Despite profound expertise and advanced surgical techniques, ischemia-induced complications ranging from wound breakdown to extensive tissue necrosis are still occurring, particularly in reconstructive flap surgery. Multiple experimental flap models have been developed to analyze underlying causes and mechanisms and to investigate treatment strategies to prevent ischemic complications. The limiting factor of most models is the lacking possibility to directly and repetitively visualize microvascular architecture and hemodynamics. The goal of the protocol was to present a well-established mouse model affiliating these before mentioned lacking elements. Harder et al. have developed a model of a musculocutaneous flap with a random perfusion pattern that undergoes acute persistent ischemia and results in ~50% necrosis after 10 days if kept untreated. With the aid of intravital epi-fluorescence microscopy, this chamber model allows repetitive visualization of morphology and hemodynamics in different regions of interest over time. Associated processes such as apoptosis, inflammation, microvascular leakage and angiogenesis can be investigated and correlated to immunohistochemical and molecular protein assays. To date, the model has proven feasibility and reproducibility in several published experimental studies investigating the effect of pre-, peri- and postconditioning of ischemically challenged tissue.

The window of the murine dorsal skinfold chamber presented visualizes a zone of acute persistent ischemia of a musculocutaneous flap. Intravital epi-fluorescence microscopy permits for direct and repetitive assessment of the microvasculature and quantification of hemodynamics. Morphologic and hemodynamic results can further be correlated with histological and molecular analyses.

在背皮褶商会鼠标的缺血性组织损伤:一个皮瓣模型，探讨急性缺血持续

Despite profound expertise and advanced surgical techniques, ischemia-induced complications ranging from wound breakdown to extensive tissue necrosis are ...

Assessment of Acute Wound Healing using the Dorsal Subcutaneous Polyvinyl Alcohol Sponge Implantation and Excisional Tail Skin Wound Models.

Wound healing is a complex process that requires the orderly progression of inflammation, granulation tissue formation, fibrosis, and resolution. Murine models provide valuable mechanistic insight into these processes; however, no single model fully addresses all aspects of the wound healing response. Instead, it is ideal to use multiple models to address the different aspects of wound healing. Here, two different methods that address diverse aspects of the wound healing response are described. In the first model, polyvinyl alcohol sponges are subcutaneously implanted along the mouse dorsum. Following sponge retrieval, cells can be isolated by mechanical disruption, and fluids can be extracted by centrifugation, thus allowing for a detailed characterization of cellular and cytokine responses in the acute wound environment. A limitation of this model is the inability to assess the rate of wound closure. For this, a tail skin excision model is utilized. In this model, a 10 mm x 3 mm rectangular piece of tail skin is excised along the dorsal surface, near the base of the tail. This model can be easily photographed for planimetric analysis to determine healing rates and can be excised for histological analysis. Both described methods can be utilized in genetically altered mouse strains, or in conjunction with models of comorbid conditions, such as diabetes, aging, or secondary infection, in order to elucidate wound healing mechanisms.

Here, two murine wound healing models are described, one designed to assess cellular and cytokine wound healing responses and the other to quantify the rate of wound closure. These methods can be used with complex disease models such as diabetes to determine mechanisms of various aspects of poor wound healing.

使用多盐皮下聚乙烯醇海绵植入和外向尾皮肤伤口模型进行急性伤口愈合评估。

Wound healing is a complex process that requires the orderly progression of inflammation, granulation tissue formation, fibrosis, and resolution. Murine ...

免疫与感染

Rat Burn Model to Study Full-Thickness Cutaneous Thermal Burn and Infection

Burn induction methodologies are inconsistently described in rat models. A uniform burn wound model, which represents the clinical scenario, is necessary to perform reproducible burn research. The present protocol describes a simple and reproducible method to create ~20% total body surface area (TBSA) full-thickness burns in rats. Here, a 22.89 cm2 (5.4 cm diameter) copper rod heated at 97&#160;&#176;C in a water bath was applied to the rat skin surface to induce the burn injury. A copper rod with a high thermal conductivity was able to dissipate the heat deeper in the skin tissue to create a full-thickness burn. Histology analysis shows attenuated epidermis with coagulative damage to the full-thickness extent of the dermis and the subcutaneous tissue. Additionally, this model is representative of the clinical situations observed in hospitalized burn patients following burn injury such as immune dysregulation and bacterial infections. The model can recapitulate the systemic bacterial infection by both Gram-positive and Gram-negative bacteria. In conclusion, this paper presents an easy-to-learn and robust rat burn model that mimics the clinical situations, including immune dysregulation and bacterial infections, which is of considerable utility for the development of new topical antibiotic drugs for burn wound and infections.

A model mimicking the clinical scenario of burn injury and infection is necessary for furthering burn research. The present protocol demonstrates a simple and reproducible rat burn infection model comparable to that in humans. This facilitates the study of burn and infections following burn for developing new topical antibiotic treatments.

大鼠烧伤模型研究全层皮肤热烧伤和感染

Burn induction methodologies are inconsistently described in rat models. A uniform burn wound model, which represents the clinical scenario, is necessary ...

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 ...

Improved Rodent Model of Myocardial Ischemia and Reperfusion Injury

Myocardial ischemia and reperfusion injury (MIRI), induced by coronary heart disease (CHD), causes damage to the cardiomyocytes. Furthermore, evidence suggests that thrombolytic therapy or primary percutaneous coronary intervention (PPCI) does not prevent reperfusion injury. There is still no ideal animal model for MIRI. This study aims to improve the MIRI model in rats to make surgery easier and more feasible. A unique method for establishing MIRI is developed by using a soft tube during a key step of the ischemic period. To explore this method, thirty rats were randomly divided into three groups: sham group (n = 10); experimental model group (n = 10); and existing model group (n = 10). Findings of triphenyltetrazolium chloride staining, electrocardiography, and percent survival are compared to determine the accuracies and survival rates of the operations. Based on the study results, it has been concluded that the improved surgery method is associated with a higher survival rate, elevated ST-T segment, and larger infarct size, which is expected to mimic the pathology of MIRI better.

Myocardial ischemia-reperfusion model of rat heart is improved by using a self-made retractor, polyvinyl chloride tube, and a unique knotting method. Electrocardiogram, triphenyltetrazolium chloride and histological staining, and percent survival analysis results showed that the improved model group has higher success and survival rates than the already existing model group.

心肌缺血再灌注损伤的改进啮齿动物模型

Myocardial ischemia and reperfusion injury (MIRI), induced by coronary heart disease (CHD), causes damage to the cardiomyocytes. Furthermore, evidence ...

A Protocol for Constructing a Rat Wound Model of Type 1 Diabetes

A single high dose of streptozotocin injection followed by full-thickness skin excision on the dorsum of rats is a common method for constructing animal models of type 1 diabetic wounds. However, improper manipulation can lead to model instability and high mortality in rats. Unfortunately, there are few existing guidelines on type 1 diabetic wound modeling, and they lack detail and do not present specific reference strategies. Therefore, this protocol details the complete procedure for constructing a type 1 diabetic wound model and analyzes the progression and angiogenic characteristics of the diabetic wounds. Type 1 diabetic wound modeling involves the following steps: preparation of the streptozotocin injection, induction of type 1 diabetes mellitus, and construction of the wound model. The wound area was measured on day 7 and day 14 after wounding, and the skin tissues of the rats were extracted for histopathological and immunofluorescence analysis. The results revealed that type 1 diabetes mellitus induced by 55 mg/kg streptozotocin was associated with lower mortality and a high success rate. The blood glucose levels were relatively stable after 5 weeks of induction. The diabetic wound healing rate was significantly lower than that of normal wounds on day 7 and day 14 (p &#60; 0.05), but both could reach more than 90% on day 14. Compared with the normal group, the epidermal layer closure of diabetic wounds on day 14 was incomplete and had delayed re-epithelialization and significantly lower angiogenesis (p &#60; 0.01). The type 1 diabetic wound model constructed based on this protocol has the characteristics of chronic wound healing, including poor closure, delayed re-epithelialization, and decreased angiogenesis compared to normal rat wounds.

The streptozotocin-induced diabetic wound model in male SD rats is currently the most widely used model for studying wound healing in type I diabetes mellitus. This protocol describes the methods used to construct this model. It also presents and addresses potential challenges and examines the progression and angiogenic characteristics of diabetic wounds.

一种构建1型糖尿病大鼠伤口模型的方案

A single high dose of streptozotocin injection followed by full-thickness skin excision on the dorsum of rats is a common method for constructing animal ...