Name: demonstration of the rat ischemic skin wound model
Uploaded: 2015-04-01T15:00:00.000+00:00
Duration: 8 min 35 s
Description: Watch this Scientific Journal Video about Demonstration of the Rat Ischemic Skin Wound Model at JoVE.com

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>

<ol>
	<li>Ansell, D. M., Holden, K. A., Hardman, M. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Animal+models+of+wound+repair:+Are+they+cutting+it.">Animal models of wound repair: Are they cutting it.</a> Experimental dermatology. 21 (8), 581-585 (2012).</li><li>Dorsett-Martin, W. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rat+models+of+skin+wound+healing:+a+review.">Rat models of skin wound healing: a review.</a> Wound repair and regeneration : official publication of the Wound Healing Society [and] the European Tissue Repair Society. 12 (6), 591-599 (2003).</li><li>Davidson, J. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Animal+models+for+wound+repair.">Animal models for wound repair.</a> Arch Dermatol Res. 290 (S1-11), (1998).</li><li>Finn, G., Magnus, S. A., Tonny, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Models+for+use+in+wound+healing+research:+A+survey+focusing+on+in+vitro+and+in+vivo+adult+soft+tissue).">Models for use in wound healing research: A survey focusing on in vitro and in vivo adult soft tissue).</a> Wound Repair and Regeneration. 8, (2000).</li><li>Gould, L. J., Leong, M., Sonstein, J., Wilson, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Optimization+and+validation+of+an+ischemic+wound+model.">Optimization and validation of an ischemic wound model.</a> Wound repair and regeneration : official publication of the Wound Healing Society [and] the European Tissue Repair Society. 13 (6), 576-582 (2004).</li><li>Loots, M. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Differences+in+cellular+infiltrate+and+extracellular+matrix+of+chronic+diabetic+and+venous+ulcers+versus+acute+wounds.">Differences in cellular infiltrate and extracellular matrix of chronic diabetic and venous ulcers versus acute wounds.</a> The Journal of investigative dermatology. 111 (5), 850-857 (1998).</li><li>Schwartz, D. A., Lindblad, W. J., Rees, R. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Altered+collagen+metabolism+and+delayed+healing+in+a+novel+model+of+ischemic+wounds.">Altered collagen metabolism and delayed healing in a novel model of ischemic wounds.</a> Wound Repair Regen. 3 (2), 204-212 (1995).</li><li>Chen, C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Molecular+and+mechanistic+validation+of+delayed+healing+rat+wounds+as+a+model+for+human+chronic+wounds.">Molecular and mechanistic validation of delayed healing rat wounds as a model for human chronic wounds.</a> Wound Repair and Regeneration. 7, (1999).</li><li>Zhang, Q., Chang, Q., Cox, R. A., Gong, X., Gould, L. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hyperbaric+oxygen+attenuates+apoptosis+and+decreases+inflammation+in+an+ischemic+wound+model.">Hyperbaric oxygen attenuates apoptosis and decreases inflammation in an ischemic wound model.</a> The Journal of investigative dermatology. 128 (8), 2102-2112 (2008).</li><li>Zhang, Q., Gould, L. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hyperbaric+oxygen+reduces+matrix+metalloproteinases+in+ischemic+wounds+through+a+redox-dependent+mechanism.">Hyperbaric oxygen reduces matrix metalloproteinases in ischemic wounds through a redox-dependent mechanism.</a> The Journal of investigative dermatology. 134 (1), 237-246 (2013).</li><li>Moor, A. N., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Consequences+of+age+on+ischemic+wound+healing+in+rats:+altered+antioxidant+activity+and+delayed+wound+closure.">Consequences of age on ischemic wound healing in rats: altered antioxidant activity and delayed wound closure.</a> Age (Dordrecht, Netherlands). 36 (2), 733-748 (2014).</li><li>Roy, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Characterization+of+a+preclinical+model+of+chronic+ischemic+wound).">Characterization of a preclinical model of chronic ischemic wound).</a> Physiological genomics. 37 (3), 211-224 (2009).</li><li>Biswas, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hypoxia+inducible+microRNA+210+attenuates+keratinocyte+proliferation+and+impairs+closure+in+a+murine+model+of+ischemic+wounds.">Hypoxia inducible microRNA 210 attenuates keratinocyte proliferation and impairs closure in a murine model of ischemic wounds.</a> Proceedings of the National Academy of Sciences of the United States of America. 107 (15), 6976-6981 (2010).</li><li>Winocour, S., Vorstenbosch, J., Trzeciak, A., Lessard, L., Philip, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=CD109,+a+novel+TGF-beta+antagonist,+decreases+fibrotic+responses+in+a+hypoxic+wound+model.">CD109, a novel TGF-beta antagonist, decreases fibrotic responses in a hypoxic wound model.</a> Exp Dermatol. 23 (7), 475-479 (2014).</li></ol>

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?...

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?...

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

Research

JoVE Journal

Medicine

21.2K 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

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 ...

科研

医学

A Murine Closed-chest Model of Myocardial Ischemia and Reperfusion

Surgical trauma by thoracotomy in open-chest models of coronary ligation induces an immune response which modifies different mechanisms involved in ischemia and reperfusion. Immune response includes cytokine expression and release or secretion of endogenous ligands of innate immune receptors. Activation of innate immunity can potentially modulate infarct size. We have modified an existing murine closed-chest model using hanging weights which could be useful for studying myocardial pre- and postconditioning and the role of innate immunity in myocardial ischemia and reperfusion. This model allows animals to recover from surgical trauma before onset of myocardial ischemia.
Volatile anesthetics have been intensely studied and their preconditioning effect for the ischemic heart is well known. However, this protective effect precludes its use in open chest models of coronary artery ligation. Thus, another advantage could be the use of the well controllable volatile anesthetics for instrumentation in a chronic closed-chest model, since their preconditioning effect lasts up to 72 hours. Chronic heart diseases with intermittent ischemia and multiple hit models are other possible applications of this model.
For the chronic closed-chest model, intubated and ventilated mice undergo a lateral blunt thoracotomy via the 4th intercostal space. Following identification of the left anterior descending a ligature is passed underneath the vessel and both suture ends are threaded through an occluder. Then, both suture ends are passed through the chest wall, knotted to form a loop and left in the subcutaneous tissue. After chest closure and recovery for 5 days, mice are anesthetized again, chest skin is reopened and hanging weights are hooked up to the loop under ECG control.
At the end of the ischemia/reperfusion protocol, hearts can be stained with TTC for infarct size assessment or undergo perfusion fixation to allow morphometric studies in addition to histology and immunohistochemistry.

Surgical trauma induces an inflammatory response. Cytokines and endogenous ligands are known to modulate myocardial infarct size following ischemia and reperfusion. We present a modified closed-chest model of murine ischemia and reperfusion using hanging weights to minimize effects of thoracotomy.

闭胸鼠心肌缺血再灌注模型

Surgical trauma by thoracotomy in open-chest models of coronary ligation induces an immune response which modifies different mechanisms involved in ...

Murine Model of Hindlimb Ischemia 

In the United States, peripheral arterial disease (PAD) affects about 10 million individuals, and is also prevalent worldwide. Medical therapies for symptomatic relief are limited. Surgical or endovascular interventions are useful for some individuals, but long-term results are often disappointing. As a result, there is a need for developing new therapies to treat PAD. The murine hindlimb ischemia preparation is a model of PAD, and is useful for testing new therapies. When compared to other models of tissue ischemia such as coronary or cerebral artery ligation, femoral artery ligation provides for a simpler model of ischemic tissue. Other advantages of this model are the ease of access to the femoral artery and low mortality rate. 
In this video, we demonstrate the methodology for the murine model of unilateral hindimb ischemia. The specific materials and procedures for creating and evaluating the model will be described, including the assessment of limb perfusion by laser Doppler imaging. This protocol can also be utilized for the transplantation and non-invasive tracking of cells, which is demonstrated by Huang et al.1.

Murine Model of Hindlimb Ischemia

The surgical procedure for induction of unilateral hindlimb ischemia is demonstrated, with confirmation of ischemia by laser Doppler perfusion imaging.

肢体缺血小鼠模型

In the United States, peripheral arterial disease (PAD) affects about 10 million individuals, and is also prevalent worldwide.  Medical therapies for ...

生物学

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 ...

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 ...

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 ...

Middle Cerebral Artery Occlusion in Rats with Preserved Vascular Anatomy

A Rat Model of Middle Cerebral Artery Occlusion/Reperfusion Without Damaging the Anatomical Structure of Cerebral Vessels

Ischemic stroke stands as the primary cause of long-term disability and mortality among adults worldwide. Animal models of ischemic stroke have significantly contributed to our understanding of its pathological mechanisms and the development of potential treatments. Presently, there are two common methods involving filament (endovascular suture) techniques to induce animal models of cerebral ischemia. However, these methods have inherent limitations, such as reduced blood perfusion to the brain, damage to the external carotid artery system, impaired food and/or water intake, and sensory dysfunction of the face. This article introduces a new method for inducing a rat ischemic stroke model without compromising the cerebral vascular anatomy. In this study, the common carotid artery (CCA) of Sprague-Dawley rats was exposed, and an incision was made. A filament was then inserted through the incision into the internal carotid artery to occlude the middle cerebral artery. After 1.5 h of induced ischemia, the occluding filament was fully removed from both the internal carotid artery and the CCA. The incision in the CCA was subsequently sutured using 11-0 microsurgical sutures under a microscope (magnification 4x). Through the utilization of microsurgical techniques to repair the CCA, this study successfully developed a unique method to induce an ischemic stroke model in rats while preserving the anatomical integrity of cerebral blood vessels.

The current protocol describes a rat model of middle cerebral artery occlusion/reperfusion that preserves the anatomical structure of cerebral vessels without causing damage.

Ischemic stroke stands as the primary cause of long-term disability and mortality among adults worldwide. Animal models of ischemic stroke have ...

神经科学

Model of Ischemia and Reperfusion Injury in Rabbits

The protocol here provides a simple, highly replicable methodology to induce in situ acute regional myocardial ischemia in the rabbit for non-survival and survival experiments. New Zealand White adult rabbit is sedated with atropine, acepromazine, butorphanol, and isoflurane. The animal is intubated and placed on mechanical ventilation. An intravenous catheter is inserted into the marginal ear vein for the infusion of medications. The animal is pre-medicated with heparin, lidocaine, and lactated Ringer&#39;s solution. A carotid cut-down is performed to obtain arterial line access for blood pressure monitoring. Select physiologic and mechanical parameters are monitored and recorded by continuous real-time analysis.
With the animal sedated and fully anesthetized, either a fourth intercostal space small left thoracotomy (survival) or midline sternotomy (non-survival) is performed. The pericardium is opened, and the left anterior descending (LAD) artery is located.
A polypropylene suture is passed around the second or third diagonal branch of the LAD artery, and the polypropylene filament is threaded through a small vinyl tube, forming a snare. The animal is subjected to 30 min of regional ischemia, achieved by occluding the LAD by tightening the snare. Myocardial ischemia is confirmed visually by regional cyanosis of the epicardium. Following regional ischemia, the ligature is loosened, and the heart is allowed to re-perfuse.
For both survival and non-survival experiments, the myocardial function can be assessed via an echocardiography (ECHO) measurement of the fractional shortening. For non-survival studies, data from sonomicrometry collected using three digital piezoelectric ultrasonic probes implanted within the ischemic area and the left ventricle developed pressure (LVDP) using an apically inserted left ventricle (LV) catheter can be continuously acquired for evaluating the regional and global myocardial function, respectively.
For survival studies, the incision is closed, a left needle thoracentesis is performed for pleural air evacuation, and postoperative pain control is achieved.

The present study demonstrates a highly reproducible animal model of acute regional myocardial ischemia and reperfusion injury in rabbits using a left mini-thoracotomy for survival cases or a midline sternotomy for non-survival cases.

兔子缺血和再灌注损伤模型

The protocol here provides a simple, highly replicable methodology to induce in situ acute regional myocardial ischemia in the rabbit for non-survival and ...