Name: demonstration of the rat ischemic skin wound model
Uploaded: 2015-04-01T15:00:00
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).

참고 : 아래에 제시된 모든 동물 절차에 대한 승인은 사우스 플로리다의 동물 관리위원회 (IACUC)의 대학에서 얻은 동물 복지법 및 실험실 동물의 관리 및 사용을위한 설명서의 모든 요구 사항을 준수했다. 실리콘 시트 및 수술 도구 1. 준비 <ol><li> 규격에 맞게 자르는 스트립 (10.5 cm X 3.0 cm) 0.01 두께, 의료용 실리콘 시트를 비는 강화 및 오토 클레이브를 사용하여 소독. </li><li> ...

<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>

<html> 쥐의 상처 치유는 종종 인해 감염된 상처와 interanimal 변화 (5)의 높은 비율을 치유 할 수있는 능력에 논쟁의 대상이되어왔다. 개발 중 모델의 원래 목표 중 하나는이 편차를 감소하는 것이었다. 특정 위치와 상처의 수를 감소 플랩의 폭에 대한 수정이, 실리콘 시트 및 소개 (일관성 cranio-꼬리 위치와 플랩을 중심으로)이 목표를 달성했다. 수축에 의해 상처 치유는 또한 인간에서와 같이,...

효과적인 약물 개발 및 다른 상처 치유 치료제는 인간의 치료 1 동물 모델에서 연구 결과를 번역하는 알려진 문제점에도 불구하고, 생체 내 모델에서 적절한 필요합니다. 다음은 병리 기전 상처 치유의 추가적인 이해를 조사하는 허혈성 피부 상처 치유 쥐 모델의 사용을위한 프로토콜의 상세한 설명이다. 쥐의 종, 종종 넓은 가용성, 크기로 인해 고용하고 절개 및 절제 상처, 이미징 및 조직 2 컬렉션에 적합한 피부 영역을 제공하기에 충분히 큰 같이 유순 자연 상처 치유 연구에 사용됩니다. 그러나 쥐가 느슨한 피부 동물 언급되면서, 쥐와 인간의 피부 해부학 적 차이가 심하게 고려해야한다. 이 독특한 특성은 래트 피부의 폐쇄 w 크게 기여하기보다는 오히려 상피화, 상처 수축 허​​용ounds 2. 또한, 쥐의 피하 panniculus 카르노 서스 근육의 존재는, 수축 모두 콜라겐 형성에 의해 3,4- 치유에 기여한다. 이러한 매우 중요한 해부학 적 차이는 허혈 래트의 피부 상처 모델의 개발을 고려하고, 특정 변형 상처 수축을 감소시키고 panniculus 카르노 서스의 근육 (5)의 영향력을 감소시키기 위해 실행되었다. 당뇨병 성 족부 궤양, 다리 궤양, 및 욕창에서 치유가 지연되고 이러한 상처는 만성 간주됩니다. 상처는 상처 수리 (6)의 다음 단계로 진행되지 않도록 상처 과도한 염증을 특징으로한다. 만성 상처의 개발에 주요 요인 중 하나는 염증을 취소 무능력에 기여 지역화 된 조직의 허혈 (감소 혈액의 흐름) 5입니다. 이 모델은 2003-4 년에 (개발 및 유효성을 검사 한 동안시), 상처 침대, 정상적인 상처 치유 동안 키 단계와이 모델 5 개발을위한 동기 부여 혈관 신생 유도를 테스트하기에 충분한 조직을 제공 할 수있는 표준화 된 동물 모델은 없었다. 즉, 여기에 제시된 모델은 첸 등에 의해 변형 된 형태로 사용 원래 슈워츠 등. 7, 이후 설명하는 허혈성 상처 모델의 변형입니다 말했다. (8) 변성 허혈성 상처 모델에서 변경 오히려 상피화보다 수축에 의해 치유 이어질 래트 상술 한 해부학 적 특징을 우회 이루어져왔다 : (1) 두 전층 절제 상처는 bipedicled 등쪽 피부 플랩 내에 생성된다 panniculus 카르노 서스 근육은 단지 근육의 근막 위의 해부에 의해 침대 상처에서 제거됩니다. (2) 자체가 플랩 t의 중간에 위치하는 더 좁은 혈액 공급이 랜덤 인 것을 보장 차원 및 상처를 가지고그는 플랩 허혈성 있습니다. (3) 실리콘 시트는 하부 조직으로부터 플랩의 재관류 재 부착을 방지 플랩 아래에 삽입된다. 상처 수축 고정 또는 실리콘 시트 (5)에 의해 봉합 플랩 (제거되지 않은) 제한된다. 이 모델은 최근 세 쥐 11 대 젊은 허혈성 상처 치유에 9,10 치유 허혈성 상처에 고압 산소의 효과에 이르기까지 연구에 사용되었으며, 장기간 조직 허혈의 신뢰할 수있는 모델로 입증되었습니다. bipedicled 플랩의 크기는 돼지 (12)와 마우스를 포함하여 다른 종 (3.0-3.5 cm 폭 10.5 cm 길이)와 F344 쥐 (2cm 폭으로 11cm 길이) 스프 라그을 포함한 다른 쥐의 변종에 맞게 조정되었습니다 13, 14. 이 비디오는 허혈성 피부 상처 모델의 데모에서 F344 근친 쥐의 피로를 사용합니다. 아래에 제시된 모든 동물 절차에 대한 승인 획득했다사우스 플로리다의 동물 관리위원회 (IACUC)의 대학에서 에드와 동물 복지법 및 관리에 대한 가이드 및 실험 동물의 사용의 모든 요구 사항을 준수.

<table border="0" cellpadding="0" cellspacing="0" width="605">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Name of Reagent/ Equipment</td>
			<td style="width:103px;">Company</td>
			<td style="width:119px;">Catalog Number</td>
			<td style="width:167px;">Comments/Description</td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:216px;">Sil-Tec medical grade sheeting</td>
			<td style="width:103px;">Technical Products Inc.</td>
			<td style="width:119px;">500-3</td>
			<td>nonreinforced, 0.01 inches in thickness</td>
		</tr>
		<tr height="61">
			<td height="61" style="height:61px;width:216px;">Mini Iris scissors, 8cm, curved, SS</td>
			<td style="width:103px;">World Precision Instruments</td>
			<td style="width:119px;">#503671</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Ethilon Nylon Sutures</td>
			<td style="width:103px;">Ethicon</td>
			<td style="width:119px;">1964G</td>
			<td>black, size 4.0, PC-3 16mm needles (3/8 circle)</td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:216px;">Laser Doppler Imager</td>
			<td style="width:103px;">Moor Instruments</td>
			<td style="width:119px;">moorLDI2-IR</td>
			<td>Standard blood flow imager: http://us.moor.co.uk/product/moorldi2-laser-doppler-imager/8</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">ImageJ</td>
			<td style="width:103px;">NIH</td>
			<td style="width:119px;">free download</td>
			<td>http://rsb.info.nih.gov/ij/</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Mastisol</td>
			<td style="width:103px;">Henry Schein</td>
			<td style="width:119px;">Cat # 7289210</td>
			<td>Fisher Scientific NC9774929</td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:216px;">Tegaderm</td>
			<td style="width:103px;">Medical Specialties&#160;</td>
			<td style="width:119px;">3M1624W</td>
			<td>Fisher Scientific NC9922128</td>
		</tr>
	</tbody>
</table>

demonstration of the rat ischemic skin wound model

인간의 만성 상처에 대한 성향 등으로 인해 부동에 당뇨병과 장애인 심혈관 기능 및 누그러 압력 노화, 질병 상태에 따라 증가한다. 동물 모델은 만성 상처의 복잡성의 이해를 발전하기위한 목적으로 이러한 조건을 모방하는 것을 시도 개발되었다. 본 명세서에 기재된 모델은 허혈성되고 만성 상처 표현형 (혈관 감소, 염증 및 지연 증가 상처 봉합)을 닮은 상처 발생 혈류 감소 연장을 허용 래트 피변 허혈 모델이다. 그것은 컨트롤 등의 플랩 측 방향의 중앙 배치의 2 허혈성 상처와이 비 허혈성 상처와 bipedicled 등의 플랩으로 구성되어 있습니다. 이 피변 허혈 모델 신규 추가 배리어 및 혈관 재 형성을 방지 및 상처 치유로 수축을 감소시키기 부목로서 기능 플랩 아래에 실리콘 시트의 배치이다. 에도 불구하고때문에 사람에 비해 자신의 매우 독특한 해부학 및 생리 학적 차이에 대한 연구를 상처 치유에 대한 쥐를 사용하는 논쟁 (즉, panniculus의 카르노 서스 근육, 짧은 수명의 존재, 모낭의 수가 증가하고, 감염된 상처를 치유 할 수있는 능력) 이 모델에 사용 된 수정은 이전에 개발 된 허혈성 피부 플랩 모델에 귀중한 대체합니다.

효율적으로 수행 한 경우 쥐의 허혈성 상처 치유 모델 프로토콜은 동물 당 약 20 분 소요됩니다. 이전 그림 1G에 나타낸 바와 같이 나타납니다 모델을 드레싱의 응용 프로그램. 그것은 bipedicled 플랩과 상처 안에 허혈성 있는지 확인하는 것이 중요 할 것입니다. 상처의 수준에서 피하 산소 장력 (PSCO 2) 두 허혈성 상처 사이의 피하 조직에 폴라 전극을 배치하여이 모델 5의...

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

쥐가 크기 때문에, 가용성, 오히려 유순 한 행동에, 몇 년 동안 연구 모델로 이용되고있다. 이 프로토콜의 목적은 만성 상처의 병태 생리에 대한 통찰력을 제공하는 허혈성 상처 치유 피부 모델로서 쥐를 이용하는 것이다.

쥐 허혈성 피부 상처 모델의 데모

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

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&amp;#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.

전체 혈액 자극 분석을 사용하여 프로 염증성 시토킨 IL - 1β의 정확하고 간단한 측정

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

연구

의학

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.

허혈성 상처 모델 생산을위한 단순화된 테크닉

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.

쥐의 레이저 도플러 Flowmetry 안내와 허혈성 뇌졸중에 대한 엔도 테린-1 유도 중간 뇌 동맥 폐색 모델

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.

허혈성 쥐 폐의 혈관 신생

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.

상처 치유의 생쥐 모델

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.

마우스의 등쪽 피하 지방 상공 회의소에서 허혈성 조직 손상 : 피부 플랩 모델은 급성 영구 허혈을 조사하기

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.

흉터 상처 수리의 마우스 태아의 피부 모델

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.

광 화학적으로 유도 된 후방 허혈성 시신경 병증의 쥐 모델

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.

신생아 저산소 - 허혈성 뇌증의 돼지 모델

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.

Nonarteritic 전방 허혈성 시신경 병증의 설치류 모델 (rNAION)

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