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?...

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 border="0" cellpadding="0" cellspacing="0" width="605">
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		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Name of Reagent/ Equipment</td>
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			<td height="41" style="height:41px;width:216px;">Sil-Tec medical grade sheeting</td>
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			<td>Standard blood flow imager: http://us.moor.co.uk/product/moorldi2-laser-doppler-imager/8</td>
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			<td height="21" style="height:21px;width:216px;">ImageJ</td>
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			<td style="width:119px;">free download</td>
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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?...

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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

21.1K 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

Accurate and Simple Measurement of the Pro-inflammatory Cytokine IL-1&beta; using a Whole Blood Stimulation Assay

Inflammatory processes resulting from the secretion of soluble mediators by immune cells, lead to various manifestations in skin, joints and other tissues as well as altered cytokine homeostasis. The innate immune system plays a crucial role in recognizing pathogens and other endogenous danger stimuli. One of the major cytokines released by innate immune cells is Interleukin (IL)-1. Therefore, we utilize a whole blood stimulation assay in order to measure the secretion of inflammatory cytokines and specifically of the pro-inflammatory cytokine IL-1&beta; 1, 2, 3.
Patients with genetic dysfunctions of the innate immune system causing autoinflammatory syndromes show an exaggerated release of mature IL-1&beta; upon stimulation with LPS alone. In order to evaluate the innate immune component of patients who present with inflammatory-associated pathologies, we use a specific immunoassay to detect cellular immune responses to pathogen-associated molecular patterns (PAMPs), such as the gram-negative bacterial endotoxin, lipopolysaccharide (LPS). These PAMPs are recognized by pathogen recognition receptors (PRRs), which are found on the cells of the innate immune system 4, 5, 6, 7. A primary signal, LPS, in conjunction with a secondary signal, ATP, is necessary for the activation of the inflammasome, a multiprotein complex that processes pro-IL-1&beta; to its mature, bioactive form 4, 5, 6, 8, 9, 10.
The whole blood assay requires minimal sample manipulation to assess cytokine production when compared to other methods that require labor intensive isolation and culturing of specific cell populations. This method differs from other whole blood stimulation assays; rather than diluting samples with a ratio of RPMI media, we perform a white blood cell count directly from diluted whole blood and therefore, stimulate a known number of white blood cells in culture 2. The results of this particular whole blood assay demonstrate a novel technique useful in elucidating patient cohorts presenting with autoinflammatory pathophysiologies.

Accurate and Simple Measurement of the Pro-inflammatory Cytokine IL-1&#946; using a Whole Blood Stimulation Assay

We describe a simple immunoassay to measure the production of pro-inflammatory cytokines, such as IL-1 beta production, in patients presenting with autoinflammatory phenotypes. By activating cells in whole blood cultures with pathogen-associated molecular patterns, specifically with lipopolysaccharide, cytokine secretion can be conveniently evaluated in whole blood supernatants.

使用全血刺激试验的促炎性细胞因子IL -1β的精确和简单的测量

Inflammatory processes resulting from the secretion of soluble mediators by immune cells, lead to various manifestations in skin, joints and other tissues ...

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 ...

Endothelin-1 Induced Middle Cerebral Artery Occlusion Model for Ischemic Stroke with Laser Doppler Flowmetry Guidance in Rat

Stroke is the number one cause of disability and third leading cause of death in the world, costing an estimated $70 billion in the United States in 20091, 2. Several models of cerebral ischemia have been developed to mimic the human condition of stroke. It has been suggested that up to 80% of all strokes result from ischemic damage in the middle cerebral artery (MCA) area3. In the early 1990s, endothelin-1 (ET-1) 4 was used to induce ischemia by applying it directly adjacent to the surface of the MCA after craniotomy. Later, this model was modified 5 by using a stereotaxic injection of ET-1 adjacent to the MCA to produce focal cerebral ischemia. The main advantages of this model include the ability to perform the procedure quickly, the ability to control artery constriction by altering the dose of ET-1 delivered, no need to manipulate the extracranial vessels supplying blood to the brain as well as gradual reperfusion rates that more closely mimics the reperfusion in humans5-7. On the other hand, the ET-1 model has disadvantages that include the need for a craniotomy, as well as higher variability in stroke volume8. This variability can be reduced with the use of laser Doppler flowmetry (LDF) to verify cerebral ischemia during ET-1 infusion. Factors that affect stroke variability include precision of infusion and the batch of the ET-1 used6. Another important consideration is that although reperfusion is a common occurrence in human stroke, the duration of occlusion for ET-1 induced MCAO may not closely mimic that of human stroke where many patients have partial reperfusion over a period of hours to days following occlusion9, 10. This protocol will describe in detail the ET-1 induced MCAO model for ischemic stroke in rats. It will also draw attention to special considerations and potential drawbacks throughout the procedure.

Several animal models of cerebral ischemia have been developed to simulate the human condition of stroke. This protocol describes the endothelin-1 (ET-1) induced middle cerebral artery occlusion (MCAO) model for ischemic stroke in rats. In addition, important considerations, advantages, and shortcomings of this model are discussed.

内皮素-1诱导大脑中动脉闭塞模型缺血性中风激光多普勒血流仪在大鼠指导

Stroke is the number one cause of disability and third leading cause of death in the world, costing an estimated $70 billion in the United States in ...

Angiogenesis in the Ischemic Rat Lung

The adult lung is perfused by both the systemic bronchial artery and the entire venous return flowing through the pulmonary arteries. In most lung pathologies, it is the smaller systemic vasculature that responds to a need for enhanced lung perfusion and shows robust neovascularization. Pulmonary vascular ischemia induced by pulmonary artery obstruction has been shown to result in rapid systemic arterial angiogenesis in man as well as in several animal models. Although the histologic assessment of the time course of bronchial artery proliferation in rats was carefully described by Weibel 1, mechanisms responsible for this organized growth of new vessels are not clear. We provide surgical details of inducing left pulmonary artery ischemia in the rat that leads to bronchial neovascularization. Quantification of the extent of angiogenesis presents an additional challenge due to the presence of the two vascular beds within the lung. Methods to determine functional angiogenesis based on labeled microsphere injections are provided.

The lung is perfused by both the systemic bronchial artery and pulmonary arteries. In most lung pathologies, it is the smaller systemic vasculature that shows robust neovascularization. Cessation of pulmonary blood flow promotes brisk bronchial angiogenesis. We provide surgical details of inducing left pulmonary artery ischemia that promotes bronchial neovascularization.

在缺血大鼠肺的血管生成

The  adult lung is perfused by both the systemic bronchial artery and the entire  venous return flowing through the pulmonary arteries. In most lung ...

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. ...

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 ...

A Mouse Fetal Skin Model of Scarless Wound Repair

Early in utero, but not in postnatal life, cutaneous wounds undergo regeneration and heal without formation of a scar. Scarless fetal wound healing occurs across species but is age dependent. The transition from a scarless to scarring phenotype occurs in the third trimester of pregnancy in humans and around embryonic day 18 (E18) in mice. However, this varies with the size of the wound with larger defects generating a scar at an earlier gestational age. The emergence of lineage tracing and other genetic tools in the mouse has opened promising new avenues for investigation of fetal scarless wound healing. However, given the inherently high rates of morbidity and premature uterine contraction associated with fetal surgery, investigations of fetal scarless wound healing in vivo require a precise and reproducible surgical model. Here we detail a reliable model of fetal scarless wound healing in the dorsum of E16.5 (scarless) and E18.5 (scarring) mouse embryos.

During mammalian development, early gestational skin wounds heal without a scar. Here we detail a reliable and reproducible model of fetal scarless wound healing in the cutaneous dorsum of E16.5 (scarless) and E18.5 (scarring) mouse embryos.

对无瘢痕修复鼠标胎儿皮肤模型

Early in utero, but not in postnatal life, cutaneous wounds undergo regeneration and heal without formation of a scar. Scarless fetal wound healing occurs ...

Rat Model of Photochemically-Induced Posterior Ischemic Optic Neuropathy

Posterior Ischemic optic neuropathy (PION) is a sight-devastating disease in clinical practice. However, its pathogenesis and natural history have&#160;remained poorly understood. Recently, we developed a reliable, reproducible animal model of PION and tested the treatment effect of some neurotrophic factors in this model1. The purpose of this video is to demonstrate our photochemically induced model of posterior ischemic optic neuropathy, and to evaluate its effects with retrograde labeling of retinal ganglion cells. Following surgical exposure of the posterior optic nerve, a photosensitizing dye, erythrosin B, is intravenously injected and a laser beam is focused onto the optic nerve surface. Photochemical interaction of erythrosin B and the laser during irradiation damages the vascular endothelium, prompting microvascular occlusion mediated by platelet thrombosis and edematous compression. The resulting ischemic injury yields a gradual but pronounced retinal ganglion cell dieback, owing to a loss of axonal input - a remote, injury-induced and clinically relevant outcome. Thus, this model provides a novel platform to study the pathophysiologic course of PION,&#160;and can be further optimized for testing therapeutic approaches for optic neuropathies as well as other CNS ischemic diseases.

The goal of this protocol is to photochemically induce ischemic injury to the posterior optic nerve in rat. This model is critical to studies of the pathophysiology of posterior ischemic optic neuropathy, and therapeutic approaches for this and other optic neuropathies, as well as of other CNS ischemic diseases.

光化学诱导后部缺血性视神经病变的大鼠模型

Posterior Ischemic optic neuropathy (PION) is a sight-devastating disease in clinical practice. However, its pathogenesis and natural history ...

A Piglet Model of Neonatal Hypoxic-Ischemic Encephalopathy

Birth asphyxia, which causes hypoxic-ischemic encephalopathy (HIE), accounts for 0.66 million deaths worldwide each year, about a quarter of the world&#8217;s 2.9 million neonatal deaths. Animal models of HIE have contributed to the understanding of the pathophysiology in HIE, and have highlighted the dynamic process that occur in brain injury due to perinatal asphyxia. Thus, animal studies have suggested a time-window for post-insult treatment strategies. Hypothermia has been tested as a treatment for HIE in pdiglet models and subsequently proven effective in clinical trials. Variations of the model have been applied in the study of adjunctive neuroprotective methods and piglet studies of xenon and melatonin have led to clinical phase I and II trials1,2. The piglet HIE model is further used for neonatal resuscitation- and hemodynamic studies as well as in investigations of cerebral hypoxia on a cellular level. However, it is a technically challenging model and variations in the protocol may result in either too mild or too severe brain injury. In this article, we demonstrate the technical procedures necessary for establishing a stable piglet model of neonatal HIE. First, the newborn piglet (&#60; 24 hr old, median weight 1500 g) is anesthetized, intubated, and monitored in a setup comparable to that found in a neonatal intensive care unit. Global hypoxia-ischemia is induced by lowering the inspiratory oxygen fraction to achieve global hypoxia, ischemia through hypotension and a flat trace amplitude integrated EEG (aEEG) indicative of cerebral hypoxia. Survival is promoted by adjusting oxygenation according to the aEEG response and blood pressure. Brain injury is quantified by histopathology and magnetic resonance imaging after 72 hr.

Hypoxic-ischemic encephalopathy following perinatal asphyxia can be studied using animal models. We demonstrate the procedures necessary for establishing a piglet model of neonatal hypoxic-ischemic encephalopathy.

新生儿缺氧缺血性脑病的小猪模型

Birth asphyxia, which causes hypoxic-ischemic encephalopathy (HIE), accounts for 0.66 million deaths worldwide each year, about a quarter of the ...

The Rodent Model of Nonarteritic Anterior Ischemic Optic Neuropathy (rNAION)

Nonarteritic anterior ischemic optic neuropathy (NAION) is a focal ischemic lesion of the optic nerve that affects 1/700 individuals throughout their lifetime. NAION results in optic nerve edema, selective loss of the retinal ganglion cell neurons (RGCs) and atrophy of the optic nerve. A rodent model of NAION that expresses most NAION features and sequelae has been developed, which is applicable to both rats and mice. This model utilizes a focal laser application of 532 nm wavelength to illuminate a photoactive dye, Rose Bengal (RB), to cause capillary damage and leakage at the targeted anterior optic nerve (the laminar region). After rNAION induction, there is an early optic nerve ischemia, optic nerve edema, and intraneural inflammation, followed by selective RGC and axonal loss. Since the optic nerve is a CNS white matter tract, the rNAION model is applicable to mechanistic studies of selective white matter ischemia, as well as neuroprotective analyses and short and long-term mechanisms of glial and neuronal response to ischemia.

The Rodent Model of Nonarteritic Anterior Ischemic Optic Neuropathy rNAION

The following report describes how to replicate the rodent model of nonarteritic anterior ischemic optic neuropathy (rNAION), using the appropriate dye, contact lens and laser parameters. We also reveal the appropriate steps for evaluating the rNAION lesion in vivo.

非动脉炎性前部缺血性视神经病变的啮齿动物模型（rNAION）

Nonarteritic anterior ischemic optic neuropathy (NAION) is a focal ischemic lesion of the optic nerve that affects 1/700 individuals throughout their ...