우리는 폐의 캐스팅에 지원 한 박사 Adlah Sukkar, MD의 작품을 인정합니다. 이 작품은 NHLBI, HL088005로 운영되었습니다.

쥐에서 수행 모든 프로토콜은 존스 홉킨스 대학 동물 관리 및 사용위원회와 NIH의 지침에 따라 승인되었습니다. 가능하면 동물은 수술 외과 사이트의 오염을 최소화하기 위해 수술 영역에서 별도의 지역에 준비해야합니다. 1. 마취 / 무통 <ol><li> 3퍼센트 isoflurane과 함께 주입 유도 챔버에서, 장소 쥐 (할렌, 인디애나 폴리스, IN Sprague Dawley 남성의 쥐, 125~150그램). </li><li> 3퍼센트 isoflurane 마취와 코 콘과 환풍기에 연결된 수술 보드에 anesthetized 쥐를 놓습니다. 모든 절차에 대한 무균 기술을 사용합니다. 멸균 운영 필드를 보장하기 위해 쥐를 내리면 t. </li></ol> 2. 삽관 법 <ol><li> 외과 테이프를 사용하여 부정사 위치에 부속을 고정. </li><li> 코 콘에서 쥐를 제거 패딩 집게로 혀를 확장합니다. </li><li> 가이드로 무딘 금속 stilet와 14 게이지 intracath을 사용합니다. 에 뒤에 슬라이드기관에 ngue. </li><li> 기관에 흰색 플라스틱 intracath을 떠날 금속 stilet를 제거합니다. 쥐 호흡하며 공기가 튜브를 통하여 흐르는 확인하십시오. </li><li> 카테터에 직접 어댑터 코 콘을 교체, 환풍기 (, 8 ML / kg 갯벌 볼륨, 설치류 환풍기 모델 683, 하버드 장치, Holliston, 매사추세츠 90 숨 / 분)에 쥐를 연결합니다. </li><li> Puralube (버틀러 Schein, 더블린, OH) 눈에 수의학 연고를 적용 할 수 있습니다. </li></ol> 3. Thoracotomy <ol><li> 장소 쥐 오른쪽 아래, 외과 테이프를 사용하여 부속을 고정. </li><li> 왼쪽 흉곽 영역을 면도. </li><li> 작은 진공과 초과 털을 제거합니다. </li><li> Povidone - 요오드 swabstick (Dynarex 공사, Orangebur, NY)에 의해 다음 알코올로 닦고하여 살균 필드를 확인합니다. 이 과정을 두 번 이상 (3 스크럽 총)을 반복합니다. </li><li> 필드의 중앙에 메스 멸균 해부 가위 또는 멸균과 횡 방향의 절개를합니다. </li><li> 리터를 해부 퉁명아래 갈비 (마지막 층은 갈비를 덮고 얇은 막입니다)에 조직과 지방의 에어. </li><li> 3 늑간 공간을 결정하기 위해 갈비뼈를 계산합니다. </li><li> 멸균 45 ° Graefe 집게를 사용하여 3, 4 번째 갈비뼈 사이의 무딘 절개를합니다. </li><li> , 갈비뼈 구분 기호를 삽입 부드럽게 개방 유지하기 위해 봉합에 열려있는 폐의 전체 시각화와 무효, 장소 테이프를 만드는 당긴다. </li></ol> 4. 좌측 폐동맥의 결합 <ol><li> 90 ° Graefe의 포셉으로 오른손으로 다시 왼쪽 폐를 이동합니다. </li><li> 뒤몽 패턴을 # 5 바로 집게를 사용하여, 왼쪽 손으로 왼쪽 폐 동맥과기도를 잡아. 왼쪽 폐동맥기도 위에 누워 것입니다. 포셉은 테이블에 직접 수직 있는지 확인합니다. 또한, 그것은 가장 말초 위치 (실질에 가장 가까운)에서 좌측 폐동맥 왼쪽 / mainstem의 기관지을 수령 것이 가장 효과적입니다. 이 왼쪽 기동과 옆 환기 왼쪽 폐를 밀어 넣습니다. </li><li> 뒤몽을 사용패턴 # 5 45 °의 자연 국경에서 왼쪽 mainstem의 기관지에서 왼쪽 폐동맥을 분리 곡선 포셉. 이 분리 라인은 두 개인 구조 사이의 얇고 흰 나타납니다. </li><li> 혈관을하지 않고 동맥에서 바로 분리, 부드럽게 분리를 따라 함께 개최 포셉 팁을 끼 웁니다. </li><li> 포셉의 팁이 가시적 왼쪽 폐 동맥과 왼쪽 mainstem의 기관지를 분리 할 때까지 계속합니다. 팁의 작은 점은 완전히 통해 있어야합니다. 혈액 포셉의 끝에서 시각화 할 수 있다면, 그것은을 통해 완전히되지 않고 결합을 시도하지 않아야합니다. 일단 공간을 통해 왼쪽 폐동맥 포셉의 곡선에 누워 것입니다. 이 위치에 잡아. </li><li> 부드럽게 직선 무딘 집게 그립 (왼쪽)을 공개하고 사전 절단 봉합 (~ 2~3인치, 폴리 프로필렌 봉합사 크기 6-0, Myco 의료, 캐리, NC)의 조각을 잡아. </li><li> 좌측 폐동맥와 g를 데리고 곡선 포셉을 엽니 다랩의 봉합. 부드럽게 포셉의 곡선에 상대적으로 상승 운동에 좌측 폐동맥과 왼쪽 mainstem의 기관지 사이의 공간을 통해 봉합사를 당겨. </li><li> 사각형 매듭과 왼쪽 폐, 봉합의주의 싹둑 나머지 부분을 occluding 아래로 묶어주세요. </li><li> (파란색 모노 필라멘트) 폴리 프로필렌으로 두 번 갈비뼈와 봉합을 보유하는 무딘 집게를 사용하여 닫기 갈비 크기주의하지로하는 것 19mm, hemostat에 바늘을 (Myco 의료, 캐리, NC) 절단 8분의 3 원 역에 첨부 4-0 봉합 피부 (단 갈비). </li><li> 느슨한 광장 매듭을 완료, 긍정적 인 엔드 - 날숨 압력 (따위가 나오기 시작, 2-5 cmH 2 O)에, 장소를 폐를 부풀려, 단단히 폐 후 안전한 매듭을 hyperinflate하고, 봉합의 나머지 부분을 snipping 전에 다른 모든 매듭을합니다. 따위가 나오기 시작에서 제거하고 폐는 붕괴하지 않도록 30 초에 표시합니다. 5 물방울 Bupivicaine을 (APP 제약, Schaumbur, IL) 적용됩니다. </li><li> 닫기 상처를 조직 접착제를 삽입하여 피부와 함께 피부를 밀어은 전용 다시 사용면 팁 작은 주걱의 D. 이후 절개의 위치 24 시간마다 8 시간이나 동물이 정상적인 활동을 다시 시작 때까지 Bupivicaine (2.0 밀리그램 / kg 피하)를 제공합니다. 이 지역의 피하 층은 매우 얇은으로, 이것 자체로는 치료해야 쉽지 않다. 조직 접착제가 적용되고 피부가 두명이 경우 또한 피하 층을 닫습니다. </li><li> isoflurane 가스를 해제하지만, 자발적인 움직임을 반환 할 때까지 실내 공기에 1-2 분 동안 쥐를 환기하고 있습니다. 환풍기에서 tracheal 튜브를 분리하고 쥐를 제거하기 전에 자발적 호흡이되어 있는지 확인합니다. </li><li> 면봉 및 모니터 운동 및 복구와 눈에서 Puralube을 닦아주십시오. (0.05 MG / kg intraperitoneal, 버틀러 Schein, 더블린 OH) buprenorphine 히드로 클로라이드를 삽입. 수술 후 48 시간에 대한 진통제 매 12 시간 배달을 계속합니다. </li></ol> 5. 왼쪽 경동맥 동맥 Cannulation 기관지 perfus의 크기를 평가하려면좌측 폐동맥의 결합 후 원하는 시간 지점에서 허혈성 왼쪽 폐의 이온은, 대동맥으로 왼쪽 경동맥을 통해 표시 마이크로를 삽입. 1-2 위와 같이 쥐를 준비합니다. <ol><li> 목을 따라 중간 선을 잘라 기관과 왼쪽 경동맥 (microsphere 분사 사이트)를 나타 내기 위해 해부 퉁명. </li><li> 25g 바늘, 4 방향 꼭지, 1 ML의 주사기에 연결된 용기로 PE20 튜브 (Becton 디킨슨, MD를 스파크)의 heparinized 생리, 무딘 팁이 가득 카테터를 삽입합니다. </li><li> 물 sonicator에서 30 초 동안, 마이크로 (Invitrogen, 유진, 또는 X 10 6 영역 / ML 15 μm 진홍의 폴리스티렌 형광 마이크로, 1) 장소 병. </li><li> 병, 소용돌이를 제거하고 20g 바늘을 통해 1 ML 해밀턴 유리 주사기 (해밀턴 회사, 리노 NV)에 0.5 ML (50 마이크로을)를 그립니다. </li><li> 4 방식 꼭지에 해밀턴 주사기를 첨부 주사기 펌프 (속도로 마이크로을 불러 일으킬 : 500 μl / 분, 지니 플러스, 켄트 과학, Torringt) 코네티컷 있습니다. </li><li> 500 μl / 분에서 heparized 생리의 1 ML과 해밀턴 주사기와 수세식 장치를 제거합니다. </li><li> 전체 가슴 thoracotomy을 수행하고 아래 대정맥을 severing하여 쥐를 ...에게서 피를 뽑다. </li><li> 왼쪽 폐와 관심의 다른 조직을 제거합니다. </li><li> 조직에서 마이크로 압축을 풀려면 일어난 과다 출혈 후 쥐의 전체 좌측 폐를 타고 2M 코 (4-6 ML)에 배치합니다. 55 ° C의 물을 욕조에 장소와 조직 소화 밤새 둡니다. , 구슬을 씻어 소용돌이 (10 초)과 원심 분리기를 (; 10 분 20 ° C 2,000 RPM) 십대 초반 80 (0.25 %)를 추가합니다. , 표면에 뜨는을 제거합니다 2 ethoxyethyl 초산 (1 ML), 소용돌이를 추가하고 1 시간에 서 보자. 소용돌이 정지 및 원심 분리기 (2,000 RPM, (20 ° C 10 분 용) 히타치 F-2500 형광 분광 광도계를 사용하여 큐벳과 방법으로 형광, 장소를 포함하는 2 ethoxyethyl 초산 수용액 층 (여기 612/emission 618를 제거합니다. , Digilab, Holliston, MA). </li></ol>

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관심 없음 충돌이 선언 없습니다.

테스트 모든 종의 좌측 폐동맥의 결합은 허혈성 폐의 강력한 조직 neovascularization로 연결됩니다. 우리는 쥐 모델에서 수술 방법의 세부 사항을 제시하고 있습니다. 우리의 결과는 혈관 주조, 조직 병리학에서 생산 및 생체 라벨에 기관지 동맥 세포 분열 따위에 의해 번식을 명시하고 폐 실질을 perfuse. 따라서, 기관지 혈관 신생의 메커니즘은 만성 폐 thromboembolism의 인간의 상태를 패러랠 동물 ...

폐의 조직 혈관 신생은 잘 인식됩니다. 이러한 천식 2, 간질 폐 섬유증이 3 암 4 및 만성 폐 thromboembolism 5의 전신 vasculature과 폐 실질 폐 proliferates 주변과 침공과 같은 질병 상태에서. 그러나, 조직의 차동 활성화보다는 폐 순환을 공부하는 동물 모델은 몇 가지 있습니다. 아마도 성인 포유 동물의 폐에 조직 neovascularization의 가장 재현 모델은 만성 폐 동맥 국소 빈혈을 유도 한 후 발생하는 것입니다. 5-7 인간 좌측 폐동맥 폐쇄에 대한 반응은, 개 8, 돼지 9, 양 10, 기니 돼지 11, 쥐 1, 12, 13 및 마우스 14 기관지 동맥뿐만 아니라 갈비 사이 동맥의 급속한 확산이다. 전신 neovasculari에 대한 책임 메커니즘폐 국소 빈혈 후 폐의 zation은 대부분 알 수없는 널리 연구되지 ​​않았습니다. 좌측 폐동맥 폐쇄 후 쥐의 기관지 혈관 신생의 시간 코스는 신중하게 Weibel 1의 histologic 작품에 설명되어 있습니다. 쥐에이 작품 확장, 저희 연구실이 과정에서 중요한 성장 요인뿐만 아니라 폐에서이 neovasculature의 physiologic 결과 모두에 초점을 맞추고있다. 결과가 CXC 케모카인의 CINC-3은 초기 국소 빈혈 후에 CXCR2에 중화 항체를 쥐를 치료 올라간다 보여, CINC-3 수용체는 혈관 신생 13 attenuates. 십사일 폐 국소 빈혈의 발병 후 새로 설립 된 기관지 vasculature 수가 크게 증가 단백질 투자율 15 이상으로 표시했습니다. 왼쪽 폐 기능은 diffusing 용량과 폐 볼륨 15 감소 감소 일반 게재되지 않았습니다. neovasculature가 공동있을 수 있습니다 있지만만성 폐 국소 빈혈 동안 폐 조직의 보존에 ntributed, 그것은 정상하고 폐 기능의 지속적인 감소에 기여할 수는 없습니다 나타납니다. 아마도이 모델의 가장 호기심 측면 중 하나는 혈관을 proliferating의 공간적 분포에 관한 것이다. 국소 빈혈로 인한 폐 실질 조직 내에서의 성장 요인의 출시에도 불구하고, neovasculature이 상대적으로 큰 업스트림 기관지 동맥에서 유래 된 것입니다. 정상적인 기관지 동맥은 대동맥에서 작은 지점으로 발생하고 카리나의기도 트리를 침공. 따라서 성장 요인 arteriogenesis의 초기 단계를 유도하는하여 메커니즘은 명확하지 않습니다. 우리는 쥐가 인간과 비슷한 혈관 해부학 적으로 폐 국소 빈혈 동안 전신 혈관 신생을 담당하는 메커니즘을 연구 할 수있는 독특한 기회를 제공하는 것이 좋습니다. 좌측 폐동맥의 완전한 방해가 인간의 과목에서 드문이지만하면,증가 기관지 vascularity는 유사 환자에서 유도 할 것으로 보인다 폐동맥 폐쇄 (16)의 어떤 사이트와 크기입니다. 따라서, 우리는 쥐의 왼쪽 폐 동맥과 혈관 신생의 정도를 수량화 할 수있는 수단을 ligate 할 수있는 수술 방법에 대한 자세한 설명을 제공합니다.

<table border="1"><tbody><tr><td> 시약 : </td><td> 회사 </td><td></td><td></td></tr><tr><td> buprenorphine 히드로 클로라이드, Puralube </td><td> 버틀러 Schein </td><td></td><td></td></tr><tr><td> bupivicaine </td><td> APP 제약 </td><td></td><td></td></tr><tr><td> Povidone - 요오드 swabstick </td><td> Dynarex 공사 </td><td></td><td></td></tr><tr><td> 폴리 프로필렌의 봉합 크기 6 대 0으로 8분의 3 원 역 절단 바늘 </td><td> Myco 의료 </td><td></td><td></td></tr><tr><td> PE20 튜브 </td><td> Becton 디킨슨 </td><td></td><td></td></tr><tr><td> 15 μm 진홍의 폴리스티렌 fluorospheres </td><td> Invitrogen </td><td></td><td></td></tr><tr><td> 한 ML 해밀턴 유리 주사기 </td><td> 해밀턴 회사 </td><td> &lt;/ TD&gt; <td></td></tr><tr><td> 장비 : </td><td></td><td></td><td></td></tr><tr><td> 램프의 요정 플러스 주사기 펌프 </td><td> 켄트 과학 </td><td></td><td></td></tr><tr><td> 형광 분광 </td><td> Digilab </td><td></td><td></td></tr><tr><td> 쥐 환풍기 모델 683 </td><td> 하버드 장치 </td><td></td><td></td></tr><tr><td></td><td></td><td></td><td> 표 1. 특정 시약 및 장비 표. </td></tr></tbody></table>

angiogenesis in the ischemic rat lung

성인 폐는 전신 기관지 동맥과 폐동맥 동맥을 통해 흐르는 전체 정맥 반환 모두에 의해 perfused 있습니다. 대부분의 폐 pathologies에서는 향상된 폐 관류에 대한 필요에 응답하고 강력한 neovascularization을 보여주는 작은 조직 vasculature입니다. 폐 동맥 방해에 의해 유도 폐 혈관 국소 빈혈은 사람의 급속한 전신 동맥 혈관 신생뿐만 아니라 여러 동물 모델에서 결과로 표시되었습니다. 쥐의 기관지 동맥 확산의 시간 코스의 histologic 평가는 신중하게 Weibel 1 설명되었지만, 새로운 혈관의 조직 성장을위한 책임이 메커니즘은 명확하지 않습니다. 우리는 기관지 neovascularization로 연결 쥐에서 좌측 폐동맥의 국소 빈혈을 일으킬 외과 세부 정보를 제공합니다. 혈관 신생의 정도의 정량화는 폐 내의 두 혈관 침대의 존재로 인해 추가 과제를 제공합니다. 방법표시 microsphere 주사에 따라 기능 혈관 신생을 결정하기 위해 제공됩니다.

혈관 캐스트 : 쥐의 좌측 폐동맥의 국소 빈혈의 효과의 결과는 그림 1에 설명되어 있습니다. 표시하면 기관지 vasculature의 메타 크릴 레이트 캐스트와 28일 LPAL 후 왼쪽기도 트리의 광범위한 vascularity입니다. 이 캐스트를 얻으려면 조직 vasculature는 내림차순으로 대동맥에 역행, 메타 크릴 레이트 혼합물 (빨간색)로 주입하였으며, 기관은 cannulated과 흰색 실리콘 기반 소재로 주입되었다. ?...

Watch this Scientific Journal Video about Angiogenesis in the Ischemic Rat Lung at JoVE.com

Angiogenesis in the Ischemic Rat Lung

폐는 전신 기관지 동맥과 폐동맥 동맥 모두에서 perfused 있습니다. 대부분의 폐 pathologies에서는 강력한 neovascularization을 보여주는 작은 조직 vasculature입니다. 폐 혈액 흐름의 중단이 활발 기관지 혈관 신생을 촉진합니다. 우리는 기관지 neovascularization을 촉진 좌측 폐동맥의 국소 빈혈을 일으킬 외과 세부 정보를 제공합니다.

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 ...

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Medicine

15.5K 조회수.  Johns Hopkins University. 폐는 전신 기관지 동맥과 폐동맥 동맥 모두에서 perfused 있습니다. 대부분의 폐 pathologies에서는 강력한 neovascularization을 보여주는 작은 조직 vasculature입니다. 폐 혈액 흐름의 중단이 활발 기관지 혈관 신생을 촉진합니다. 우리는 기관지 neovascularization을 촉진 좌측 폐동맥의 국소 빈혈을 일으킬 외과 세부 정보를 제공합니다.

비디오: Angiogenesis in the Ischemic Rat Lung

High-frequency High-resolution Echocardiography: First Evidence on Non-invasive Repeated Measure of Myocardial Strain, Contractility, and Mitral Regurgitation in the Ischemia-reperfused Murine Heart

Ischemia-reperfusion (IR) was surgically performed in murine hearts which were then subjected to repeated imaging to monitor temporal changes in functional parameters of key clinical significance. Two-dimensional movies were acquired at high frame rate (8 kHz) and were utilized to estimate high-quality myocardial strain. Two-dimensional elastograms (strain images), as well as strain profiles, were visualized. Results were powerful in quantitatively assessing IR-induced changes in cardiac events including left-ventricular (LV) contraction, LV relaxation and isovolumetric phases of both pre-IR and post-IR beating hearts in intact mice. In addition, compromised sector-wise wall motion and anatomical deformation in the infarcted myocardium were visualized.  The elastograms were uniquely able to provide information on the following parameters in addition to standard physiological indices that are known to be affected by myocardial infarction in the mouse: internal diameters of mitral valve orifice and aorta, effective regurgitant orifice, myocardial strain (circumferential as well as radial), turbulence in blood flow pattern as revealed by the color Doppler movies and velocity profiles, asynchrony in LV sector, and changes in the length and direction of vectors demonstrating slower and asymmetrical wall movement. This work emphasizes on the visual demonstration of how such analyses are performed. 

High-frequency High-resolution Echocardiography: First Evidence on Non-invasive Repeated Measure of Myocardial Strain, Contractility, and Mitral Regurgitation in the Ischemia-reperfused Murine Heart 

High frequency Doppler ultrasound is a novel technology for assessing regional myocardial function. This work presents first evidence demonstrating applicability of this versatile imaging platform for the repeated measure of myocardial strain, dp/dt, and mitral regurgitation in the ischemia-reperfused (IR) murine heart.

고주파 고해상도 Echocardiography : 국소 빈혈 - reperfused Murine 중심부에 심근 스트레인, 수축성 및 승모판 병리 현상의 비침습 반복 측정에 대한 최초의 증거

Ischemia-reperfusion (IR) was surgically performed in murine hearts which were then subjected to repeated imaging to monitor temporal changes in ...

연구

의학

Multi-modal Imaging of Angiogenesis in a Nude Rat Model of Breast Cancer Bone Metastasis Using Magnetic Resonance Imaging, Volumetric Computed Tomography and Ultrasound

Angiogenesis is an essential feature of cancer growth and metastasis formation. In bone metastasis, angiogenic factors are pivotal for tumor cell proliferation in the bone marrow cavity as well as for interaction of tumor and bone cells resulting in local bone destruction. Our aim was to develop a model of experimental bone metastasis that allows in vivo assessment of angiogenesis in skeletal lesions using non-invasive imaging techniques.
For this purpose, we injected 105 MDA-MB-231 human breast cancer cells into the superficial epigastric artery, which precludes the growth of metastases in body areas other than the respective hind leg1. Following 25-30 days after tumor cell inoculation, site-specific bone metastases develop, restricted to the distal femur, proximal tibia and proximal fibula1. Morphological and functional aspects of angiogenesis can be investigated longitudinally in bone metastases using magnetic resonance imaging (MRI), volumetric computed tomography (VCT) and ultrasound (US).
MRI displays morphologic information on the soft tissue part of bone metastases that is initially confined to the bone marrow cavity and subsequently exceeds cortical bone while progressing. Using dynamic contrast-enhanced MRI (DCE-MRI) functional data including regional blood volume, perfusion and vessel permeability can be obtained and quantified2-4. Bone destruction is captured in high resolution using morphological VCT imaging. Complementary to MRI findings, osteolytic lesions can be located adjacent to sites of intramedullary tumor growth. After contrast agent application, VCT angiography reveals the macrovessel architecture in bone metastases in high resolution, and DCE-VCT enables insight in the microcirculation of these lesions5,6. US is applicable to assess morphological and functional features from skeletal lesions due to local osteolysis of cortical bone. Using B-mode and Doppler techniques, structure and perfusion of the soft tissue metastases can be evaluated, respectively. DCE-US allows for real-time imaging of vascularization in bone metastases after injection of microbubbles7.
In conclusion, in a model of site-specific breast cancer bone metastases multi-modal imaging techniques including MRI, VCT and US offer complementary information on morphology and functional parameters of angiogenesis in these skeletal lesions.

In the pathogenesis of bone metastasis, angiogenesis is a crucial process and therefore represents a target for imaging and therapy. Here, we present a rat model of site-specific breast cancer bone metastasis and describe strategies to non-invasively image angiogenesis in vivo using magnetic resonance imaging, volumetric computed tomography and ultrasound.

자기 공명 영상, 체적 계산의 Tomography 및 초음파를 이용한 유방암의 뼈 전이의 누드 쥐 모델에서 Angiogenesis의 멀티 모달 이미징

Angiogenesis is an essential feature of cancer growth and metastasis formation. In bone metastasis, angiogenic factors are pivotal for tumor cell ...

Retinal Detachment Model in Rodents by Subretinal Injection of Sodium Hyaluronate

Subretinal injection of sodium hyaluronate is a widely accepted method of inducing retinal detachment (RD). However, the height and duration of RD or the occurrence of subretinal hemorrhage can affect photoreceptor cell death in the detached retina. Hence, it is advantageous to create reproducible RDs without subretinal hemorrhage for evaluating photoreceptor cell death. We modified a previously reported method to create bullous and persistent RDs in a reproducible location with rare occurrence of subretinal hemorrhage. The critical step of this modified method is the creation of a self-sealing scleral incision, which can prevent leakage of sodium hyaluronate after injection into the subretinal space. To make the self-sealing scleral incision, a scleral tunnel is created, followed by scleral penetration into the choroid with a 30 G needle. Although choroidal hemorrhage may occur during this step, astriction with a surgical spear reduces the rate of choroidal hemorrhage. This method allows a more reproducible and reliable model of photoreceptor death in diseases that involve RD such as rhegmatogenous RD, retinopathy of prematurity, diabetic retinopathy, central serous chorioretinopathy, and age-related macular degeneration (AMD).

Creating experimental retinal detachments with a reproducible and sustained height of detachment, and without subretinal hemorrhage, is important for studying the pathophysiology of photoreceptor cell loss in retinal disease and evaluating potential therapeutic interventions. Here, we report such a method in detail.

나트륨 히알루 론산의 망막 주사 설치류에있는 망막 박리 모델

Subretinal injection of sodium hyaluronate is a widely accepted method of inducing retinal detachment (RD). However, the height and duration of RD or the ...

Implantation of Fibrin Gel on Mouse Lung to Study Lung-specific Angiogenesis

Recent significant advances in stem cell research and bioengineering techniques have made great progress in utilizing biomaterials to regenerate and repair damage in simple tissues in the orthopedic and periodontal fields. However, attempts to regenerate the structures and functions of more complex three-dimensional (3D) organs such as lungs have not been very successful because the biological processes of organ regeneration have not been well explored. It is becoming clear that angiogenesis, the formation of new blood vessels, plays key roles in organ regeneration. Newly formed vasculatures not only deliver oxygen, nutrients and various cell components that are required for organ regeneration but also provide instructive signals to the regenerating local tissues. Therefore, to successfully regenerate lungs in an adult, it is necessary to recapitulate the lung-specific microenvironments in which angiogenesis drives regeneration of local lung tissues. Although conventional in vivo angiogenesis assays, such as subcutaneous implantation of extracellular matrix (ECM)-rich hydrogels (e.g., fibrin or collagen gels or Matrigel - ECM protein mixture secreted by Engelbreth-Holm-Swarm mouse sarcoma cells), are extensively utilized to explore the general mechanisms of angiogenesis, lung-specific angiogenesis has not been well characterized because methods for orthotopic implantation of biomaterials in the lung have not been well established. The goal of this protocol is to introduce a unique method to implant fibrin gel on the lung surface of living adult mouse, allowing for the successful recapitulation of host lung-derived angiogenesis inside the gel. This approach enables researchers to explore the mechanisms by which the lung-specific microenvironment controls angiogenesis and alveolar regeneration in both normal and pathological conditions. Since implanted biomaterials release and supply physical and chemical signals to adjacent lung tissues, implantation of these biomaterials on diseased lung can potentially normalize the adjacent diseased tissues, enabling researchers to develop new therapeutic approaches for various types of lung diseases.

Recapitulation of the organ-specific microenvironment, which stimulates local angiogenesis, is indispensable for successful regeneration of damaged tissues. This report demonstrates a novel method to implant fibrin gels on the lung surface of living mouse in order to explore how the lung-specific microenvironment modulates angiogenesis and alveolar regeneration in adult mouse.

마우스 폐에 섬유소 젤의 주입은 폐 관련 신생 혈관 연구에

Recent significant advances in stem cell research and bioengineering techniques have made great progress in utilizing biomaterials to regenerate and ...

Method of Isolated Ex Vivo Lung Perfusion in a Rat Model: Lessons Learned from Developing a Rat EVLP Program

The number of acceptable donor lungs available for lung transplantation is severely limited due to poor quality. Ex-Vivo Lung Perfusion (EVLP) has allowed lung transplantation in humans to become more readily available by enabling the ability to assess organs and expand the donor pool. As this technology expands and improves, the ability to potentially evaluate and improve the quality of substandard lungs prior to transplant is a critical need. In order to more rigorously evaluate these approaches, a reproducible animal model needs to be established that would allow for testing of improved techniques and management of the donated lungs as well as to the lung-transplant recipient. In addition, an EVLP animal model of associated pathologies, e.g., ventilation induced lung injury (VILI), would provide a novel method to evaluate treatments for these pathologies. Here, we describe the development of a rat EVLP lung program and refinements to this method that allow for a reproducible model for future expansion. We also describe the application of this EVLP system to model VILI in rat lungs. The goal is to provide the research community with key information and &#8220;pearls of wisdom&#8221;/techniques that arose from trial and error and are critical to establishing an EVLP system that is robust and reproducible.

Method of Isolated Ex Vivo Lung Perfusion in a Rat Model: Lessons Learned from Developing a Rat EVLP Program

Ex-Vivo Lung Perfusion (EVLP) has allowed lung transplantation in humans to become more readily available by enabling the ability to assess organs and expand the donor pool. Here, we describe the development of a rat EVLP program and refinements that allow for a reproducible model for future expansion.

고립의 방법<em&gt; 전의 VIVO</em쥐 모델에서&gt; 폐 관류 : 쥐 EVLP 프로그램 개발에서 교훈을

The number of acceptable donor lungs available for lung transplantation is severely limited due to poor quality. Ex-Vivo Lung Perfusion (EVLP) has allowed ...

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 ...

Rat Heterotopic Abdominal Heart/Single-lung Transplantation in a Volume-loaded Configuration

Herein, we describe a novel technique for heterotopic abdominal heart-lung transplantation (HAHLT) in rats. The configuration of the transplant graft involves anastomosis of donor inferior vena cava (IVC) to recipient IVC, and donor ascending aorta (Ao) to recipient abdominal Ao. The right upper and middle lung lobes are preserved and function as conduits for blood flow from right heart to left heart.
There are several advantages to using this technique, and it lends itself to a broad range of applications. Because the graft is transplanted in a configuration that allows for dyamic volume-loading, cardiac function may be directly assessed in vivo. The use of pressure-volume conductance catheters permits characterization of load-dependent and load-independent hemodynamic parameters. The graft may be converted to a loaded configuration by applying a clamp to the recipient&#8217;s infra-hepatic IVC. We describe modified surgical techniques for both donor and recipient operations, and an ideal myocardial protection strategy. Depending on the experimental aim, this model may be adapted for use in both acute and chronic studies of graft function, immunologic status, and variable ventricular loading conditions. The conducting airways to the transplanted lung are preserved, and allow for acute lung re-ventilation. This facilitates analysis of the effects of the mixed venous and arterial blood providing coronary perfusion to the graft.
A limitation of this model is its technical complexity. There is a significant learning curve for new operators, who should ideally be mentored in the technique. A surgical training background is advantageous for those wishing to apply this model. Despite its complexity, we aim to present the model in a clear and easily applicable format. Because of the physiologic similarity of this model to orthotopic transplantation, and its broad range of study applications, the effort invested in learning the technique is likely to be worthwhile.

We describe a novel technique for heterotopic abdominal heart-lung transplantation (HAHLT) in rats. The transplant configuration results in a partially loaded graft circulation, allowing direct functional assessment. This model may be employed for acute or chronic studies of function and immunologic status of the transplanted graft.

쥐 이소성 복부 심장 / 볼륨로드 구성에서 단일 폐 이식

Herein, we describe a novel technique for heterotopic abdominal heart-lung transplantation (HAHLT) in rats. The configuration of the transplant graft ...

Demonstration of the Rat Ischemic Skin Wound Model

The propensity for chronic wounds in humans increases with ageing, disease conditions such as diabetes and impaired cardiovascular function, and unrelieved pressure due to immobility. Animal models have been developed that attempt to mimic these conditions for the purpose of furthering our understanding of the complexity of chronic wounds. The model described herein is a rat ischemic skin flap model that permits a prolonged reduction of blood flow resulting in wounds that become ischemic and resemble a chronic wound phenotype (reduced vascularization, increased inflammation and delayed wound closure). It consists of a bipedicled dorsal flap with 2 ischemic wounds placed centrally and 2 non-ischemic wounds lateral to the flap as controls. A novel addition to this ischemic skin flap model is the placement of a silicone sheet beneath the flap that functions as a barrier and a splint to prevent revascularization and reduce contraction as the wounds heal. Despite the debate of using rats for wound healing studies due to their quite distinct anatomic and physiologic differences compared to humans (i.e., the presence of a panniculus carnosus muscle, short life-span, increased number of hair follicles, and their ability to heal infected wounds) the modifications employed in this model make it a valuable alternative to previously developed ischemic skin flap models.

The rat, due to its size, availability, and rather docile behavior, has been utilized as a research model for many years. The goal of this protocol is to utilize the rat as an ischemic skin wound healing model to provide valuable insight into the pathophysiology of chronic wounds.

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 ...

Assessing Therapeutic Angiogenesis in a Murine Model of Hindlimb Ischemia

Critical limb ischemia (CLI) is a serious condition that entails a high risk of lower limb amputation. Despite revascularization being the gold-standard therapy, a considerable number of CLI patients are not suited for either surgical or endovascular revascularization. Angiogenic therapies are emerging as an option for these patients but are currently still under investigation. Before application in humans, those therapies must be tested in animal models and its mechanisms must be clearly understood. An animal model of hindlimb ischemia (HLI) has been developed by the ligation and excision of the distal external iliac and femoral arteries and veins in mice. A comprehensive panel of tests was assembled to assess the effects of ischemia and putative angiogenic therapies at functional, histologic and molecular levels. Laser Doppler was used for the flow measurement and functional assessment of perfusion. Tissue response was evaluated by the analysis of capillary density after staining with the anti-CD31 antibody on histological sections of gastrocnemius muscle and by measurement of collateral vessel density after diaphonization. Expression of angiogenic genes was quantified by RT-PCR targeting selected angiogenic factors exclusively in endothelial cells (ECs) after laser capture microdissection from mice gastrocnemius muscles. These methods were sensitive in identifying differences between ischemic and non-ischemic limbs and between treated and non-treated limbs. This protocol provides a reproducible model of CLI and a framework for testing angiogenic therapies.

Here, a critical hindlimb ischemia experimental model is presented followed by a battery of functional, histologic and molecular tests&#160;to assess the effectiveness of angiogenic therapies.&#8203;

힌들보 리 허 혈의 뮤린 모델에서 치료 혈관 신생 평가

Critical limb ischemia (CLI) is a serious condition that entails a high risk of lower limb amputation. Despite revascularization being the gold-standard ...

In Vitro Model of Coronary Angiogenesis

Here, we describe an in vitro culture assay to study coronary angiogenesis. Coronary vessels feed the heart muscle and are of clinical importance. Defects in these vessels represent severe health risks such as in atherosclerosis, which can lead to myocardial infarctions and heart failures in patients. Consequently, coronary artery disease is one of the leading causes of death worldwide. Despite its clinical importance, relatively little progress has been made on how to regenerate damaged coronary arteries. Nevertheless, recent progress has been made in understanding the cellular origin and differentiation pathways of coronary vessel development. The advent of tools and technologies that allow researchers to fluorescently label progenitor cells, follow their fate, and visualize progenies in vivo have been instrumental in understanding coronary vessel development. In vivo studies are valuable, but have limitations in terms of speed, accessibility, and flexibility in experimental design. Alternatively, accurate in vitro models of coronary angiogenesis can circumvent these limitations and allow researchers to interrogate important biological questions with speed and flexibility. The lack of appropriate in vitro model systems may have hindered the progress in understanding the cellular and molecular mechanisms of coronary vessel growth. Here, we describe an in vitro culture system to grow coronary vessels from the sinus venosus (SV) and endocardium (Endo), the two progenitor tissues from which many of the coronary vessels arise. We also confirmed that the cultures accurately recapitulate some of the known in vivo mechanisms. For instance, we show that the angiogenic sprouts in culture from SV downregulate COUP-TFII expression similar to what is observed in vivo. In addition, we show that VEGF-A, a well-known angiogenic factor in vivo, robustly stimulates angiogenesis from both the SV and Endo cultures. Collectively, we have devised an accurate in vitro culture model to study coronary angiogenesis.

In vitro models of coronary angiogenesis can be utilized for the discovery of the cellular and molecular mechanisms of coronary angiogenesis. In vitro explant cultures of sinus venosus and endocardium tissues show robust growth in response to VEGF-A and display a similar pattern of COUP-TFII expression as in vivo.

관상 동맥 혈관 신생의 체외 모델

Here, we describe an in vitro culture assay to study coronary angiogenesis. Coronary vessels feed the heart muscle and are of clinical importance. Defects ...