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> 청소하고 (수술 중 멸균 필드를 만들 가위, 집게와 커튼이나 수건) 적절한 수술 도구를 소독. </li></ol> 2. 실험 동물 <ol><li> 성인 남성 또는 상업적 개종에서 얻은 250-350g 범위에 무게 암컷 쥐를 사용합니다. 세 래트가 이용되는 경우, 더 잘 수술 후 생존을 보장하기 위해 ≥350 g이어야한다. 모든 실험의 시작에 앞서 적어도 모든 동물을 순응먹이와 물은 마음대로 먹게하면서 12 시간 명암주기의 표준 조건 하에서 칠일. </li></ol> 3. 마취, 수술 전 진통과 수술 준비 <ol><li> 유도 챔버를 통해 3 % -4 %를 이소 플루 란을 사용하여 전신 마취를 유도하고 피부 준비 및 수술 중 O 2 1 %에서 (코 콘의 사용을 통해) -2 %를 유지한다. 속도와 호흡, 인터 ​​핀치 또는 눈꺼풀 깜빡임과 깊이의 관찰에 의해 마취의 깊이를 모니터링합니다. 주 :이 때, 수의사 연고 동물이 마취 상태에서 건조를 방지하기 위해 눈 상에 배치 될 수있다. </li><li> 무균 수술 영역에서 위치 원격에서 발생하기 쉬운 위치에 쥐를 배치하고 목 아래 약 11cm의 기지에서 가위로 배부 면도. 영구 마커 스텐실, 3.0 cm X 10.5 cm 플랩에 대한 개요는 (그림 1A 참조). </li><li> 깨끗하고 지정된 surgica에 쥐를 이동승인 된 가열 패드 및 무균 수술 커튼이나 수건을 갖추고 리터 영역입니다. 통증 관리를위한 제 절개 수술 5 ㎎ / kg 케토 프로 피하 주사 종래. 필요에 따라 추가의 유체 (생리 식염수)를 피하에 (최대 5를 CC)을들 수있다. </li><li> 멸균 필드를 만들 멸균 커튼을 적용 후, 70 % 이소 프로필 알코올 0.2 % 클로르헥시딘과 초 먼저 보라고하여 더 피부를 준비합니다. 10 % 포비돈 - 요오드 (베타 딘)도 사용할 수있다. 참고 : 항생제 (15 ㎎ / ㎏에서 암피실린)를 피하 투여 할 수 있지만, 좋은 무균 기술을 사용하는 경우, 그것은 필요하지 않습니다. </li></ol> 4. 절제 상처와 Bipedicled 플랩 만들기 <ol><li> 멸균 일회용 6mm 생검 펀치 도구를 사용하여, 지정된 영역 플랩 (도 1b)의 중심에 두 개의 원형 &quot;허혈&quot;상처를 만든다. 상처의 깊이는 아래 (안 통해)에 panniculus의 carno의 기본 근막해야SUS 근육 (그림 1B 인세 트). </li><li> 사용 집게 펀치 생검에 의해 생성 된 상처 윤곽의 중간에 피부를 들어 올린 다음 (panniculus의 카르노 서스 근육 포함) 조직의 원형 조각을 절제하는 (곡선 팁) 홍채 가위를 사용합니다. 결과는 상처의 기부로서 근막 권취 전층 것이다. 참고 : 적출 한 조직 (상처 플러그) 액체 질소에 냉동 또는 제어와 같은 후 처리, 정상 피부에 10 % 완충 포르말린 O / N에 고정 스냅 할 수 있습니다. </li><li> 떨어져 길이 10.5 cm 3.0 cm있는 사전으로 그린 선 (그림 1C)을 따라 허혈성 상처의 각 측면에 멸균 메스로 절개를함으로써 bipedicled 플랩을 만듭니다. 절개의 깊이는 아래 paraspinous 근육해야한다. 홍채 가위를 사용하여, (6mm 펀치의 &quot;기본&quot;으로 그대로의 밴드되지 않도록 조심스럽게 paraspinous 근육에서 F를 panniculus의 카르노 서스 근막을 분리igure 1D). </li><li> 1 멸균 프리 컷 실리콘 시트를 가지고 panniculus의 카르노 서스 근막 시트가 구부러 지거나 접하지 않는 것을 보장 paraspinous 근육 (그림 1E) 사이를에 배치합니다. 블랙, 비 흡수성 봉합사 (크기 4.0)를 사용하여 플랩 (도 1F 및 1G)의 길이를 따라, 각각의 측면상의 적어도 8 중단 바늘 피부에 실리콘 시트를 고정함으로써 두 절개를 닫는다. </li><li> 멸균 일회용 생검 펀치 도구를 사용하여, 1cm가 플랩 (그림 1G)의 양쪽에있는 허혈성 상처에 대해 측면 (panniculus의 카르노 서스 근육의 앞쪽 근막 아래로)이 내부 통제 &quot;비 허혈성&quot;상처를 만들 수 있습니다. </li><li> 상처 아래 통치자를 놓고 상처 측정을 목적으로 (그림 3A 참조) 디지털 사진을 찍어. 이때, 혈류 (재관류) 레이저 도플러 또는 다른 조작이 수행 (국소 약물 배치)를 이용하여 모니터링 할 수있다.</li><li> 뇌와 꼬리 상처와 촉촉하고 (멸균) 깨끗한 상처 환경을 유지하기 위해 드레싱 투명 필름을 모두 승인 된 액체 접착제를 적용합니다. 추가적인 드레싱 가장 꼬리 봉합사를 제거하는 동물을 방지 플랩의 꼬리 단부에 배치 될 수있다. </li></ol> 5. 수술 후 절차 <ol><li> 공급 문지르는 수술 부위를 방지하기 위해 얕은 공급기가 장착 케이지 (단독으로 보관)에있는 동물을 놓습니다. 동물 방치 또는 흉골 드러 누움을 유지하고 목적이있는 운동을 전시 할 수있는 충분한 의식을 되 찾을 때까지 다른 동물의 회사에 반환 할 수 없습니다. 난방 매트를 복구하는 동안 최대 2 일 동안 케이지의 절반 아래에 배치해야합니다. </li><li> 수술 후 통증을 관리하려면, 최대 48 시간의 수술 후는 다음 아침과 하루에 1 배 동물에 피하 케토 프로 펜 (5 ㎎ / ㎏)을 관리 할 수​​ 있습니다. 동물도 매일 모니터링해야통증, 체중 감소 또는 수술 부위 감염의 징후 연장. </li></ol> 6. 후속 상처 측정 및 드레싱 변경 <ol><li> 유도 챔버를 통해 3 %에서 이소 플루 란을 사용하여 전신 마취 자주 -4 %를 허혈성 및 비 허혈성 상처를 측정하고 단계 3.1에서와 같이 O 2와 1 % -2 %에서 (코 콘을 통해) 유지. </li><li> 피부 접착제를 잡아 당기지하기로 부드럽게 드레싱을 제거합니다. 추가 디지털 사진 감아 측정에 찍은이 때, 국소 치료 재인가, 레이저 도플러 영상 (LDI) 또는 연구자의 필요에 맞게 다른 조작을 수행. </li><li> 접착제 깨끗한 드레싱을 적용하고 동물이 단계 5.1에서와 같이 복구 할 수 있습니다. </li></ol> 7. 상처 수집 및 안락사 <ol><li> 동물 공업을 통해 3 % -4 %를 이소 플루 란을 사용하여 전신 마취 상태에서 수확 허혈성 및 비 허혈성 상처 (날이 조사는 적절하다고 판단)uction 챔버 및 단계 3.1에서와 같이 O 2와 1 % -2 %에서 (코 콘을 통해) 유지. </li><li> 메스를 사용하여, 상처 주위의 건강한 조직을 일부 포함하는 상처 주위에 사각형 모양의 절개를 만든다. 1.5 ml의에 절제를 놓고 캡 튜브 스냅와 미래의 분자 분석 (-80 ° C에서 저장) 액체 질소에 동결 스냅 또는 조직 학적 처리를 위해 RT에서 10 % 포르말린의 O에서 / N을 배양한다. 주 : 상처 절개도 분석을 위해 더 많은 샘플을 제공하기 위해 반으로 잘라 될 수있다. </li><li> 상처 조직을 제거한 후, CO 2 흡입의 승인 된 방법을 사용하여 동물을 안락사. </li></ol>

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The authors have nothing to disclose.

<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><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

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

21.3K 조회수.  University of South Florida.  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. 

비디오: Demonstration of the Rat Ischemic Skin Wound Model

Acute Myocardial Infarction in Rats

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

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

고양이에서 급성 심근 경색

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

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 ...

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. ...

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.

Myocutaneous 국소 빈혈 reperfusion 상해의 쥐 모델 : 현장 가로 직근 abdominis의 Myocutaneous 플랩의

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.

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 ...

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

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

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

등지성 폴리비닐 알코올 스폰지 이식 및 절제 꼬리 피부 상처 모델을 사용하여 급성 상처 치유의 평가.

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

면역학 및 감염병학

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

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

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

전층 피부 열 화상 및 감염을 연구하기 위한 쥐 화상 모델

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

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

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

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

재건 미세 수술에서 허혈 재관류 손상의 연구를위한 전 임상 쥐 모델

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.

심근 허혈 및 재관류 손상의 개선 된 설치류 모델

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

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

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

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

제1형 당뇨병의 쥐 상처 모델을 구축하기 위한 프로토콜

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