저자는 보스턴, MA 초안을 교정하기위한 교정 및 원고를 편집하고 Paskus 예레미야, 보스턴 아동 병원, 대한, 로리 폰 Melchner, 베른 대학교 의과 대학 신경 외과학 교실, 베른, 스위스 감사합니다. 우리는 다니엘 메틀, DVM, 최대 뮐러, DVM, 다니엘 Zalokar 및 Olgica Beslac, 실험 외과 학회, 임상 연구학과, 베른 대학, 베른, 스위스 동물 보호, 마취 및 수술 지원의 숙련 된 관리를 주셔서 감사합니다. 우리는 생리 학적 매개 변수의 실시간 데이터 모니터링 및 사후 처리를 위해, 마이클 Lensch, 헤드 연구 간호사, 중환자 의학 교실, 베른 대학 병원 베른 대학, 베른, 스위스 감사합니다. 우리는 우수한 실험실 기술 및 수술 지원, 에딘 Nevzati, 칼 Muroi, 그리고 살로메 Erhardt 감사합니다. 이 작품은 인텐시브의 부서에 의해 지원되었다전자 케어 의학, 베른 대학 병원 베른, 베른, 스위스, 임상 연구학과, 베른, 베른, 스위스의 대학의 대학 및 Kantonsspital 아라우, 아라우, 스위스에서 연구 기금. 우리는 그림 1과 2에 대한 무단 전재 허가를, 엘스 비어 감사합니다.

2.5 무​​게 3 개월 된 여성 뉴질랜드 토끼 - 3.5 kg이 절차를 사용 하였다. 이 연구는 실험 동물의 관리 및 사용을위한 건강 지침의 국립 연구소에 따라 스위스, 베른 (승인 번호 13분의 105)의 광저우의 동물 관리위원회의 승인을 수행 하였다. 모든 수술은 베른, 스위스 베른 대학 병원에서 임상 연구학과의 실험 외과 연구소에서 무균 조건 하에서 수행 하였다. 동물 용 마취는 수술 중 및 복구에 걸쳐 동물을 모니터링. 1 동물 준비, 위치 및 쇄골 하 동맥 삽관 <ol><li> 근육 마취제의 주입 토끼에서 전신 마취를 유도 (30 ㎎ / ㎏, Ketalar, 50 ㎎ / ㎖) 및 자일 라진 (kg 6 밀리그램 /; Xylapan 20 ㎎ / ㎖) 유해에 토끼의 반응을 확인하여 마취의 컨트롤 깊이 stimulat이온 (예를 들면, 발가락 핀치). 긍정적 인 반응의 경우에는 1.7를 참조하십시오. </li><li> 양안의 하부 뚜껑 풀다운 건조 더욱 자극을 막기 위해 연고 눈꺼풀에 소량의 적용. </li><li> , 20 Gbutterfly의 venflon (20 G 혈관 카테터)와 측면 귀 정맥 카테 테르를 꽂다 접착 테이프로 고정하고, 0.9 %의 염화나트륨을 포함하는 중력 가방 (500 ㎖ / 24 시간)와 케타민을 연결 (40 ㎎ / ㎏ / 시간) 연속 정맥 마취 / 자일 라진 (xylazine) (4 ㎎ / ㎏ / 시간). 추가 진통제마다 15 분 IV (펜타닐, 1 MCG / kg)을 관리합니다. 참고 : CPP 감소와 연관된 휘발성 가스 마취제, CBF 증가하고, 산소 뇌 대사율의 감소를 피 11 정맥 마취제가 뇌 공부를 할 때 맨 중요하다 CBF 및 뇌 혈관 수축, 12을 유지하여 neuroanesthesia 더 이상적인 특성을 제공한다. 혈관 경련. 또한, 비록 사망률의 증가 spontaneously 동물을 호흡, 더 나은 급성 SAH의 인간의 상황을 모방 할 수 있습니다. </li><li> 호 기말 이산화탄소를 할 수있는 마스크를 통해 - (2 L / 분 1) (ETCO 2) 모니터링 산소를 제공합니다. </li><li> 토끼의 복부 측면에있는 삼각형 배열에서 3 채널 심전도 (ECG) .Place 세 피하 전극을 설치합니다 특히, 두 팔다리에 분포 하복부에 두 개의 전극 (쇄골 하 동맥 삽관에 대한 멸균 면도 필드 거리) 오른쪽 midthoracic 영역에 하나의 전극을 배치합니다. </li><li> 호흡, ECG 신호에서 모니터링 심박수 (HR), 유해 자극에 반응에 따라 마취의 깊이에게 수술 중 매 15 분을 모니터링합니다. </li><li> 유해 자극 (발가락 핀치)에 긍정적 인 응답의 경우에, 케타민 러스 (6 밀리그램 / kg) IV 및 자일 라진 러스 (0.05 밀리그램 / kg) IV 및 / 또는 펜타닐과 추가적인 진통제의 볼 루스 (1 MC함으로써 마취 깊이를 적응g / kg) IV </li><li> 아래로 머리를 20 °를 기울 쇄골 하 동맥이 노출 될에 약간 옆으로 반대측으로 돌려, 몸 온난화 접시에 앙와위에서 토끼를 수정합니다. </li><li> 눈 연고를 적용하고 쇄골의 중간 셋째 주위에 오른쪽 가슴 근육 위에 머리를 면도로 수술 영역을 준비하고, frontal- parietal- 및 후두부 두개골, 목을 통해, 오른쪽 대퇴 동맥을 통해. </li><li> 살균 넓은 스펙트럼, 예를 들면., 포비돈 요오드로 3 분 동안 피부를 소독. </li><li> 멸균 시트와 토끼를 커버. 용기의 조작 및 지역 감염에 의한 동맥 혈관 경련을 방지하기 위해 국소 멸균 조건에서 모든 추가 절차를 수행하고 자주 염산 및 항생제 용액 (네오 마이신 황산염 5 ㎎ / ㎖) papaverin 4 %를 적용합니다. </li><li> 국소 마취제 (리도카인 1 % 최대 6 ㎎ / ㎏)과 가슴 근육을 침투. 흉골 피부 절개를하고 준비가슴 근육. 현미경을 사용하여, 쇄골 하 동맥을 해부 노출 끝 주위 근위 및 원위 합자 (4-0 polyfilament 봉합) 고정합니다. 카테터를 확보하고 말단 혈관을 결찰하는 장소에 인접 제어에 가까운 일 합자를 유지합니다. </li><li> 곡선 microscissor와 동맥을 절개하여 쇄골 하 동맥의 벽에 arteriotomy을 수행하고 작은 intravasal 3 방향 스톱 콕이 역행 쇄골 하 동맥을 cannulate. 동맥 비틀림 또는 동맥의 근위 부분의 굴곡을 방지하기위한 미끄러짐 또는 출혈을 방지하기 위해 원위 봉합사를 향해 이중 매듭 합자 의해 카테터를 고정. </li></ol> 2 혈압 및 동맥 혈액 가스 모니터링 <ol><li> 내가 3 방향 스톱 콕) intravasal 동맥혈 가스 캐 뉼러 (ABG) 분석, 산도, 파코 2, PAO 2, 수소 나트륨, 기본 초과하고, SO 2를 연결하고, ⅱ) 침략 동맥혈압력 측정 장치, 및 ⅲ) 션트 장치. </li><li> ABG 상태에 대한 혈액 샘플 (파코 2, PAO 2)를 수집하고 지속적으로 아날로그 - 표준 심혈관 및 호흡기 매개 변수 (혈압, HR, ECG, 호흡 및 기말 CO 2) 아날로그 출력 인터페이스를 통해 및 전송 데이터를 모니터링 디지털 컨버터 / 데이터 로거 및 저장. NOTE : 압력은 전후 각 세션 후에 심장 수준 제로화되고 시리즈가 시작되기 전에, 아날로그 / 디지털 변환기와 데이터 로깅 시스템의 압력 조정이 한번 수행 될 것이다. </li></ol> 3 기준 디지털 감산 혈관 조영술 <ol><li> 혈관 조영술을 교정하기 위해 모두 하악 각도를 통해 외부 크기 조정 장치 (작은 영역)을 놓습니다. 참고 :이 기준의 사후 측정의 정확한 비교를 할 수 있으며 혈관 직경을 따릅니다. </li><li> 역행 내에서 디지털 감산 혈관 조영술 (DSA)을 수행비 이온 Iopamidol rterial의 일시 주사를 통한 동맥 삽관 (0.6 ㎖ / kg, 2 초 동안 5 ㎖ / 초)와 후자의 폐색을 방지하기 위해 식염수 일시 주사 직후 뉼러를 플러시. </li><li> 5 ° 왼쪽 전방 경사 위치에서 빠른 연속 혈관 조영 촬영에 vertebrobasilar 시스템의 이미지 (14 초에서 7 이미지)를 얻습니다. </li><li> 국소 마취제 (리도카인 1 %, 최대 6 ㎎ / ㎏)와 오른쪽 대퇴 동맥 주위를 침투. 작은 사타구니의 피부 절개를합니다. 시각화 현미경을 사용하여, 대퇴 동맥을 해부 노출 끝 주위 근위 및 원위 합자 (4-0 polyfilament 봉합) 고정합니다. </li><li> arteriotomy 후, 5-F 칼집으로 대퇴 동맥을 cannulate. 식염수 시스의 측면 포트를 세척. </li><li> 투시 하에서 시스를 통해 완두 동맥에 5-F 카테터를 전진. 로드맵을 만든 다음 토륨 가이드 와이어를 전진전자 vertebrobasilar 시스템. 단계 3.2에 설명 된대로 DSA에 대한 (0.6 ㎖ / kg, 2 초에서 5 ㎖ / 초) 비 이온 Iopamidol의 루스를 주입한다. </li></ol> 수 그린 위치로 4 회전 <ol><li> 기준 DSA 다음, 앙와위에서 발생하기 쉬운 위치로 토끼의 위치를​​ 변경. 조작 또는 동맥 카테터의 위치를​​ 재배치하지 않도록주의하십시오. </li><li> 머리를 아래를 지향, 30 ° 각도로 머리 홀더 머리를 놓습니다. </li></ol> 5 Cisterna 마그나 관통 <ol><li> 1 분마다 포비돈 요오드의 3 배와 머리와 목을 통해 피부를 소독, 멸균 시트와 수술 영역을 커버. </li><li> 사전 피부 절개 나 근육의 변위없이 transcutaneously cisterna 마그나에 22 G X 40mm 소아 척추 액세스 바늘을 삽입합니다. </li><li> 동물이 완전히 뼈 외부 후두 융기를 따라 아래로 바늘을 밀기 전에 발가락 - 핀치 응답의 부족을 보장함으로써 마취되어 있는지 확인갭이 검출 될 때까지; 더 바늘을 밀어하지 않습니다. </li><li> 몇 분 동안 20 ° 각도로 아래로 기울어 진 토끼의 머리와 뇌척수액의 자연 떨어지는 관찰하여 바늘의 정확한 위치를 확인합니다. </li></ol> 두개 내 압력 및 대뇌 혈액 흐름 모니터링의 6 설치 <ol><li> 중간 선 피부와 머리 덮개 절개 한 후, 작은 수술 견인기를 삽입합니다. </li><li> neuromonitoring 장치 경우의 배치 후각 망울 및 양자 정면 위에 세 라운드 절골술 (2 mm 직경) 외 두개골 도시 따른 두개골의 정면 부분에서 고속 microdrill를 사용하여 (도 1) (9), 즉, 확인 필요. 다음과 같이 버 구멍 배치를위한 좌표를 결정하기 위해 밀리미터 스케일자를 사용 : midpupillary 라인에 두개 내압 (ICP) 모니터링, midsagittal 줄에서 1~2밀리미터; 실질 내출혈 레이저 도플러 프로브4~5mm 전방 및 브레 그마에 횡 방향 (그림 1). </li><li> 경질을 시각화하고, 세심한 지혈을 수행의 압전 조치 덕분에 뼈 지혈을 위해 뼈 왁스를 사용하여 경질의 바이폴라 응고를 사용하여 로컬 지혈을 수행합니다. </li><li> 2mm의 깊이에 대한 권리 후각 망울로 실질 내출혈 두개 내압 (ICP) 모니터 팁을 배치하고 교정. </li><li> 외부 클램프 시스템을 사용하여 2 개의 레이저 도플러 flowmetry 미세 바늘 프로브를 배치하고, 좌우 정면 반구 모두 뇌척수액에 방해가되지 않도록 중앙선에서 심실 시스템, 즉 측 방향에서 대응하는 버어 구멍에 삽입. 2.5 mm의 깊이로 바늘 프로브를 놓습니다. </li><li> neuromonitoring 프로브를 배치 한 후에, 두개골 유체 기밀을 유지하기 위해 뼈 왁스의 두께 플러그 버어 모든 구멍을 밀봉. </li><li> 평균 동맥 혈압의 파라미터를 측정 기준선 (MAP), ICP 및 다중 매개 모니터와 네 채널 레이저 도플러 조직 혈류 모니터를 사용하여 뇌 혈류 (CBF). </li></ol> 7 션트 유도 <ol><li> 혈액 충전 압력 모니터링 튜브를 통해 이전에 catheterized 쇄골 하 동맥 cisterna 마그나에서 척추 액세스 바늘을 연결합니다. 혈압 측정을위한 혈액 샘플링 포트와 3 방향 스톱 콕을 사용한다. NOTE : SAH의 심각성은 혈액의 양에 의존하고, 대략 5,11 뇌 수확시 지주막 응고 extensiveness 의해 추정 될 수있다. </li><li> 지속적으로 SAH 후 최소 20 분까지 전 6 분에서 1 Hz의 샘플링 속도에서 MAP, HR, ECG, 호흡과 호 기말 이산화탄소를 모니터링 할 수 있습니다. </li><li> 동물이 완전히 쇄골 하 동맥과 cisterna 마그나 압력 gradie에 의해 SAH을 유도 사이의 병렬 연결을 열기 전에 발가락 핀치 응답의 부족을 보장함으로써 마취되어 있는지 확인NT. NOTE : (. 예 ICP의 원하는 수준으로) 제어 된 SAH는 어느 시점에서 션트를 폐쇄함으로써 성취 될 수있다. </li><li> 약 15 분의 기간 동안 정상 값을 기록한다. </li><li> ICP가 절정에 도달 한 후 ICP가 기준 값에 가까운 정상 상태로 돌아올 때까지 그 자리에 척추 액세스 바늘을 유지합니다. ICP 고원은 10 개 이상의 초 동안 유지 또는 ICP가 자발적으로 감소하면, 션트를 닫됩니다. </li><li> (연속 출혈 카테터 조작 높은 이환율과 사망률 및 감염 속도를 증가되기 때문에, 쇄골 카테터를 포함하여) 모든 카테터를 제거, 뼈 왁스와 버 구멍을 연결, CBF 미세 바늘 프로브 및 ICP 프로브를 제거와 엄격한 상처 관개를 수행 네오 마이신 황산염과 피부를 봉합. </li></ol> (8) 수술 후 관리 <ol><li> 절차는 약 2 시간 동안 지속된다. 자일 라진 및 케타민 인해, 애니의 회복 시간의 반감기에약 1 시간 - 리터는 매우 짧다. 동물은 복구 중에 가열 램프 아래에 보관됩니다. 추가 유체는 제공되지 않습니다. 초기 수술 후 회복 단계에서, 부 프레 노르 핀 0.02 밀리그램 / kg SC 24 시간마다 8 시간이 적용됩니다. </li><li> 다음 72 시간 동안 진통 효과에 대한 동물의 면도 목 지역에서 12.5 μg / 시간을 방출 경피 펜타닐 매트릭스 패치를 적용합니다. </li><li> 이 흉골 드러 누움을 유지하기 위해 충분한 의식을 회복 할 때까지 무인 동물을 두지 마십시오. </li><li> 완전히 회복 될 때까지 다른 동물의 회사에서 수술을했다 동물을 반환하지 않습니다. </li></ol> 3 일에 DCVS을 평가하기 위해 디지털 감산 혈관 조영술을 따라 9. <ol><li> 상술 한 바와 같이 3.6 단계 1.1를 수행합니다. </li><li> 나트륨 thiopenthal의 동맥 내 일시 주사 (40 ㎎ / ㎏)에 의해 동물을 안락사 (펜 토탈, Ospedalia AG, Hünenberg, 스위스). 경우 조직 학적 및 immunohist에서ochemistry 필요하다 100cm의 H 2 O의 관류 압력 RT에서 심장 내 관류 고정을 수행 </li></ol>

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None. The authors have no financial or commercial interest in any of the drugs, materials, or equipment used. No specific funding was received for this work. The authors are solely responsible for the design and conduct of the presented study and report no conflict of interest concerning the materials and methods used in this study or the findings specified in the paper. They confirm the adherence of ethical standards. The study was performed in accordance with the National Institutes of Health guidelines for the care and use of experimental animals and with the approval of the Animal Care Committee of the Canton of Bern, Switzerland (approval #109/07 and #107/09).

션트 모델 급성 SAH 3,8,10 후에 인간에서 관찰되는 것과 유사한 병리 현상을 생성한다. 그것은 EBI는 악화 유지하고 심지어 DCVS 12 트리거 등이 모델 SAH를 다음 EBI와 DCVS 상호 작용을 포함하여 초기와 후기 DCVS 단계를 모두 조사에 도움이 될 수로 수 있음을 제안하고있다. 특히, 반복 생체 DCVS DSA (13)을 포함하여 모니터링 기술, 15 개 readibly 작은 실험 동물에 비해 ...

동맥류 성 지주막 하 출혈 (SAH)은 자주 영구적 인 신경 손상 또는 사망 한 선도, 신경 병리학 적 조건을 위협하는 가장 삶의 하나입니다. 과거의 연구는 SAH 2와 관련된 신경 학적 결손의 주요 원인으로 지연 뇌 혈관 연축 (DCVS)에 초점을 맞추고있다. 그러나, 혈관 경련의 치료 후 SAH 고통받는 환자의 일반적 나쁨 임상 결과는 SAH 3 초 후 뇌 손상 (EBI)의 효과를 포함하는 연구 초점이 팽창되었다. SAH 후 나쁨 임상 결과에 기여하고 EBI DCVS 모두의 중요성은 더 큰 이해 효과적인 치료 전략의 개발이 필수적이다. 지금까지, cisterna 마그나에 단일 및 이중자가 혈액 주입 DCVS 2-6의 연구를위한 SAH 유도를위한 표준 방법이었다. 일반적으로 이전의 연구에서 사용했지만,이 모델은 대부분 SAH와 관련된 신경 병리학 주요 변경 사항을 재생하지 못하는 EBI 7을 유도. 대조적으로, 혈관 천공 부분적 EBI (7)의 증상을 모방 중증 급성 병리 생리 학적 변화를 생성하는 것으로 알려져있다. 이 보고서는 SAH의 신규 한 토끼 모델이 기재된 방법으로, 표준 cisterna 마그나 모델 동맥 시스템을 연결하여 구성된다. 이에 SAH 유도 병리학 8-10의 더 정확한 특성화를 허용 EBI 및 DCVS 양쪽의 조사를 가능하게하기위한 설명 체외 션트를 통해 쇄골 하 동맥과 cisterna 마그나의. 혈류시켜 토끼의 생리학에 링크와 동맥혈 및 두개 내압 사이의 압력 구배에 의해 구동된다. 뇌내 압력 (ICP)는 이완기 혈압이 같고 션트 시스템에서 혈액이 응고 할 때 출혈이 멈 춥니 다. 호스트 &amp; #을 활용8217;의 생리 안정적으로 EBI와 DCVS 표현형 3,8-10 모두를 생산 SAH의 일관성 모델에지도 심사관에 의존 SAH 유도를 줄일 수 있습니다.

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			<td style="width:236px;">Zeiss, Jena, Germany</td>
			<td colspan="2">Zeiss, OPMI-MD surgical microscope</td>
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			<td style="width:236px;">Camino Multi-Parameter Monitor, Integra, Plainsboro, NJ, US</td>
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			<td style="width:236px;">BIOPAC Systems, Inc., Goleta, CA, USA</td>
			<td style="width:223px;">Biopac MP100 and acqKnowledge software,version 3.8.1</td>
			<td></td>
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			<td height="60" style="height:60px;">software analysis</td>
			<td style="width:236px;">ImagePro Discovery, MediaCybernetics, Silver Spring, MD, USA</td>
			<td style="width:223px;">image-Pro Plus version&#160;</td>
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			<td height="20" style="height:20px;">angiography apparatus</td>
			<td style="width:236px;">DFP 2000 A-Toshiba</td>
			<td style="width:223px;">MIIXR0001EAA</td>
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			<td>B. Braun, Germay</td>
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			<td>GE Medical Systems, Switzerland&#160;</td>
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			<td height="40" style="height:40px;">5.5F three-lumen central venous catheter&#160;</td>
			<td style="width:236px;">Connectors, Tagelswangen, Switzerland</td>
			<td>silicone catheter STH-C040</td>
			<td></td>
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			<td height="20" style="height:20px;">22Gx40mm needle&#160;</td>
			<td style="width:236px;">Emergo Group Inc., Netherlands</td>
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			<td></td>
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			<td height="20" style="height:20px;">high-speed microdrill</td>
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			<td>5400-15&#160;</td>
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			<td style="width:236px;">Aesculap, Inc., PA, US</td>
			<td>US349SP&#160;</td>
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			<td style="width:223px;"></td>
			<td>Any generic product</td>
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			<td style="width:223px;"></td>
			<td>Any generic product</td>
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			<td style="width:223px;"></td>
			<td>Any generic product</td>
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			<td height="40" style="height:40px;">Neomycin sulfate&#160;</td>
			<td style="width:236px;">Research Organics Inc., OH, USA</td>
			<td style="width:223px;"></td>
			<td>Any generic product</td>
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			<td>Any generic product</td>
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			<td>Any generic product</td>
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			<td height="20" style="height:20px;width:279px;">Iopamidol&#160;</td>
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			<td>Any generic product</td>
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		<tr height="20">
			<td height="20" style="height:20px;width:279px;">3-0 resorbable suture</td>
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		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:279px;">5-0 non absorbable suture</td>
			<td style="width:236px;">Ethicon Inc., USA</td>
			<td>8618G</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:279px;">4-0 polyfilament sutures</td>
			<td style="width:236px;">Ethicon Inc., USA</td>
			<td style="width:223px;">VCP284G</td>
			<td></td>
		</tr>
	</tbody>
</table>

the rabbit blood-shunt model for the study of acute and late sequelae of subarachnoid hemorrhage: technical aspects

초기 뇌 손상 및 지연 뇌 혈관 경련은 모두 지주막 하 출혈 (SAH) 후 불리한 결과에 기여한다. 두 조건을 시뮬레이션 재현 및 제어 동물 모델은 현재 드물다. 따라서, 새로운 모델은 SAH로 인한 인간의 병태 생리 학적 조건을 모방하기 위해 필요하다. 이 보고서는 뇌내 압력 (ICP)의 제어를 가능하게 토끼 피 션트 SAH 모델의 기술적 인 뉘앙스를 설명합니다. 체외 션트는 동맥 폐쇄 시스템 및 두개골에 시험관 독립적 SAH있게 막밑 공간 사이에 배치된다. 단계별 절차 명령들 및 필요한 장비를 기술뿐만 아니라, 기술적 고려 최소 사망률과 질병률과 모델을 생성한다. 이 강력한 간단하고 일관성있는 ICP-제어 SAH 토끼 모델의 성공적인 수술 작성에 필요한 중요한 세부 사항은 설명한다.

<html> SAH의 토끼 피 션트 모델은이 보고서에서 설명하는 것은, 기저 피질 (그림 2A, B) 및 대뇌 혈관 (그림 2C)와 같은 초 24로 시간 부상 후 (B 그림 2A) 해마에서 EBI를 생산하고 특성을 나타낸다 혈액 분포 (그림 2D) 8. 또한,이 모델은 SAH 유도 (그림 3) 10 일 이후 세에 DCVS의 심한 정도가 적당한 트리거합니다. 급성 SAH시 호흡 ?...

Watch this Scientific Journal Video about The Rabbit Blood-shunt Model for the Study of Acute and Late Sequelae of Subarachnoid Hemorrhage: Technical Aspects at JoVE.com

The Rabbit Blood-shunt Model for the Study of Acute and Late Sequelae of Subarachnoid Hemorrhage: Technical Aspects

The experimental intracranial pressure-controlled blood shunt subarachnoid hemorrhage (SAH) model in the rabbit combines the standard procedures &#8212; subclavian artery cannulation and transcutaneous cisterna magna puncture, which enables close mimicking of human pathophysiological conditions after SAH. We present step-by-step instructions and discuss key surgical points for successful experimental SAH creation.

지주막 하 출혈의 급성과 늦은 후유증의 연구에 대한 토끼 피 션트 모델 : 기술적 측면

Early brain injury and delayed cerebral vasospasm both contribute to unfavorable outcomes after subarachnoid hemorrhage (SAH). Reproducible and ...

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13.5K 조회수.  University and Bern University Hospital (Inselspital). The experimental intracranial pressure-controlled blood shunt subarachnoid hemorrhage (SAH) model in the rabbit combines the standard procedures — subclavian artery cannulation and transcutaneous cisterna magna puncture, which enables close mimicking of human pathophysiological conditions after SAH. We present step-by-step instructions and discuss key surgical points for successful experimental SAH creation.

비디오: The Rabbit Blood-shunt Model for the Study of Acute and Late Sequelae of Subarachnoid Hemorrhage: Technical Aspects

Microsurgical Venous Pouch Arterial-Bifurcation Aneurysms in the Rabbit Model: Technical Aspects

For ruptured human cerebral aneurysms endovascular embolization has become an equivalent alternative to aneurysm clipping.1 However, large clinical trials have shown disappointing long-term results with unacceptable high rates of aneurysm recanalization and delayed aneurysm rupture.2 To overcome these problems, animal experimental studies are crucial for the development of better endovascular devices.3-5
Several animal models in rats, rabbits, canines and swine are available.6-8 Comparisons of the different animal models showed the superiority of the rabbit model with regard to hemodynamics and comparability of the coagulation system and cost-effectiveness.9-11 
The venous pouch arterial bifurcation model in rabbits is formed by a venous pouch sutured into an artificially created true bifurcation of both common carotid arteries (CCA). The main advantage of this model are true bifurcational hemodynamics.12 The major drawbacks are the sofar high microsurgical technical demands and high morbidity and mortality rates of up to 50%.13 These limitations have resulted in less frequent use of this aneurysm model in the recent years. These shortcomings could be overcome with improved surgical procedures and modified peri- and postoperative analgetic management and anticoagulation.14-16 Our techniques reported in this paper demonstrate this optimized technique for microsurgical creation of arterial bifurcation aneurysms.

An optimized technique for the microsurgical creation of arterial bifurcation aneurysms mimicking bifurcation human cerebral aneurysms is described. A venous pouch is sutured into an artificially created true bifurcation of both common carotid arteries. Facilitated microsurgical techniques and aggressive postoperative anticoagulation and analgesia lead to minimized morbidity rates and high aneurysm patency rates.

토끼 모델에서 Microsurgical 정맥 주머니 아티 - 분기점의 동맥류 : 기술적인 관점

For ruptured human cerebral aneurysms endovascular embolization has become an equivalent alternative to aneurysm clipping.1 However, large clinical trials ...

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의학

Biomarkers in an Animal Model for Revealing Neural, Hematologic, and Behavioral Correlates of PTSD

Identification of biomarkers representing the evolution of the pathophysiology of Post Traumatic Stress Disorder (PTSD) is vitally important, not only for objective diagnosis but also for the evaluation of therapeutic efficacy and resilience to trauma. Ongoing research is directed at identifying molecular biomarkers for PTSD, including traumatic stress induced proteins, transcriptomes, genomic variances and genetic modulators, using biologic samples from subjects' blood, saliva, urine, and postmortem brain tissues. However, the correlation of these biomarker molecules in peripheral or postmortem samples to altered brain functions associated with psychiatric symptoms in PTSD remains unresolved. Here, we present an animal model of PTSD in which both peripheral blood and central brain biomarkers, as well as behavioral phenotype, can be collected and measured, thus providing the needed correlation of the central biomarkers of PTSD, which are mechanistic and pathognomonic but cannot be collected from people, with the peripheral biomarkers and behavioral phenotypes, which can.
Our animal model of PTSD employs restraint and tail shocks repeated for three continuous days - the inescapable tail-shock model (ITS) in rats. This ITS model mimics the pathophysiology of PTSD 17, 7, 4, 10. We and others have verified that the ITS model induces behavioral and neurobiological alterations similar to those found in PTSD subjects 17, 7, 10, 9. Specifically, these stressed rats exhibit (1) a delayed and exaggerated startle response appearing several days after stressor cessation, which given the compressed time scale of the rat's life compared to a humans, corresponds to the one to three months delay of symptoms in PTSD patients (DSM-IV-TR PTSD Criterian D/E 13), (2) enhanced plasma corticosterone (CORT) for several days, indicating compromise of the hypothalamopituitary axis (HPA), and (3) retarded body weight gain after stressor cessation, indicating dysfunction of metabolic regulation.
The experimental paradigms employed for this model are: (1) a learned helplessness paradigm in the rat assayed by measurement of acoustic startle response (ASR) and a charting of body mass; (2) microdissection of the rat brain into regions and nuclei; (3) enzyme-linked immunosorbent assay (ELISA) for blood levels of CORT; (4) a gene expression microarray plus related bioinformatics tools 18. This microarray, dubbed rMNChip, focuses on mitochondrial and mitochondria-related nuclear genes in the rat so as to specifically address the neuronal bioenergetics hypothesized to be involved in PTSD.

We describe a rat model of post traumatic stress disorder (PTSD) that reveals the persistent alterations in neuroendocrine function and the delayed long-term, exaggerated fear response, characteristic of PTSD patients. The animal model and methods described here are useful for correlating biomarkers in brain nuclei, which are mechanistic but cannot be measured in patients, with biomarkers in peripheral white blood cells, which can.

PTSD의, 신경 Hematologic, 그리고 행동 상관 관계를 누설하는 행위에 대해서는 동물 모델에서 생체

Identification of biomarkers representing the evolution of the pathophysiology of Post Traumatic Stress Disorder (PTSD) is vitally important, not only for ...

NADH Fluorescence Imaging of Isolated Biventricular Working Rabbit Hearts

Since its inception by Langendorff1, the isolated perfused heart remains a prominent tool for studying cardiac physiology2. However, it is not well-suited for studies of cardiac metabolism, which require the heart to perform work within the context of physiologic preload and afterload pressures. Neely introduced modifications to the Langendorff technique to establish appropriate left ventricular (LV) preload and afterload pressures3. The model is known as the isolated LV working heart model and has been used extensively to study LV performance and metabolism4-6. This model, however, does not provide a properly loaded right ventricle (RV). Demmy et al. first reported a biventricular model as a modification of the LV working heart model7, 8. They found that stroke volume, cardiac output, and pressure development improved in hearts converted from working LV mode to biventricular working mode8. A properly loaded RV also diminishes abnormal pressure gradients across the septum to improve septal function. Biventricular working hearts have been shown to maintain aortic output, pulmonary flow, mean aortic pressure, heart rate, and myocardial ATP levels for up to 3 hours8.
When studying the metabolic effects of myocardial injury, such as ischemia, it is often necessary to identify the location of the affected tissue. This can be done by imaging the fluorescence of NADH (the reduced form of nicotinamide adenine dinucleotide)9-11, a coenzyme found in large quantities in the mitochondria. NADH fluorescence (fNADH) displays a near linearly inverse relationship with local oxygen concentration12 and provides a measure of mitochondrial redox state13. fNADH imaging during hypoxic and ischemic conditions has been used as a dye-free method to identify hypoxic regions14, 15 and to monitor the progression of hypoxic conditions over time10.
The objective of the method is to monitor the mitochondrial redox state of biventricular working hearts during protocols that alter the rate of myocyte metabolism or induce hypoxia or create a combination of the two. Hearts from New Zealand white rabbits were connected to a biventricular working heart system (Hugo Sachs Elektronik) and perfused with modified Krebs-Henseleit solution16 at 37 &deg;C. Aortic, LV, pulmonary artery, and left &amp; right atrial pressures were recorded. Electrical activity was measured using a monophasic action potential electrode. To image fNADH, light from a mercury lamp was filtered (350&plusmn;25 nm) and used to illuminate the epicardium. Emitted light was filtered (460&plusmn;20 nm) and imaged using a CCD camera. Changes in the epicardial fNADH of biventricular working hearts during different pacing rates are presented. The combination of the heart model and fNADH imaging provides a new and valuable experimental tool for studying acute cardiac pathologies within the context of realistic physiological conditions.

The objective is to monitor the mitochondrial redox state of isolated hearts within the context of physiologic preload and afterload pressures. A biventricular working rabbit heart model is presented. High spatiotemporal resolution fluorescence imaging of NADH is used to monitor the mitochondrial redox state of epicardial tissue.

절연 Biventricular의 NADH 형광 이미징은 토끼 하트 워킹

Since its inception by Langendorff1, the isolated perfused heart remains a prominent tool for studying cardiac physiology2. However, it is not well-suited ...

A Low Mortality Rat Model to Assess Delayed Cerebral Vasospasm After Experimental Subarachnoid Hemorrhage

Objective: To characterize and establish a reproducible model that demonstrates delayed cerebral vasospasm after aneurysmal subarachnoid hemorrhage (SAH) in rats, in order to identify the initiating events, pathophysiological changes and potential targets for treatment.
Methods: Twenty-eight male Sprague-Dawley rats (250 - 300 g) were arbitrarily assigned to one of two groups - SAH or saline control. Rat subarachnoid hemorrhage in the SAH group (n=15) was induced by double injection of autologous blood, 48 hr apart, into the cisterna magna. Similarly, normal saline (n=13) was injected into the cisterna magna of the saline control group. Rats were sacrificed on day five after the second blood injection and the brains were preserved for histological analysis. The degree of vasospasm was measured using sections of the basilar artery, by measuring the internal luminal cross sectional area using NIH Image-J software. The significance was tested using Tukey/Kramer's statistical analysis.
Results: After analysis of histological sections, basilar artery luminal cross sectional area were smaller in the SAH than in the saline group, consistent with cerebral vasospasm in the former group. In the SAH group, basilar artery internal area (.056 &mu;m &plusmn; 3) were significantly smaller from vasospasm five days after the second blood injection (seven days after the initial blood injection), compared to the saline control group with internal area (.069 &plusmn; 3; p=0.004). There were no mortalities from cerebral vasospasm.
Conclusion: The rat double SAH model induces a mild, survivable, basilar artery vasospasm that can be used to study the pathophysiological mechanisms of cerebral vasospasm in a small animal model. A low and acceptable mortality rate is a significant criterion to be satisfied for an ideal SAH animal model so that the mechanisms of vasospasm can be elucidated 7, 8. Further modifications of the model can be made to adjust for increased severity of vasospasm and neurological exams.

Aneurysmal subarachnoid hemorrhage (SAH) is bleeding that occurs into the subarachnoid space when an aneurysm ruptures. While the morbidity and mortality from this event has been on a decline due to improved treatment approaches, the risk of vasospasm after subarachnoid hemorrhage continues to be the same as it was several years ago. The importance of establishing a comprehensive and reproducible animal model to identify initiating events of cerebral vasospasm has been the focus of research since the first use of rats in an experimental vasospasm model in 1979 by Barry et al. Early work in rats demonstrated that a single injection of autologous blood into the cisterna magna led to acute (within minutes) but not delayed cerebral vasospasm 3, 6, 14. Here we characterize a low mortality SAH rat model that results in reproducible delayed vasospasm.

실험 Subarachnoid의 출혈 후 뇌 지연 Vasospasm을 평가하기 위해 낮은 사망 쥐 모델

Objective: To characterize and establish a reproducible model that demonstrates delayed cerebral vasospasm after aneurysmal subarachnoid hemorrhage (SAH) ...

Human Neuroendocrine Tumor Cell Lines as a Three-Dimensional Model for the Study of Human Neuroendocrine Tumor Therapy

Neuroendocrine tumors (NETs) are rare tumors, with an incidence of two per 100,&#x200A;000 individuals per year, and they account for 0.5% of all human malignancies.1 Other than surgery for the minority of patients who present with localized disease, there is little or no survival benefit of systemic therapy. Therefore, there is a great need to better understand the biology of NETs, and in particular define new therapeutic targets for patients with nonresectable or metastatic neuroendocrine tumors. 3D cell culture is becoming a popular method for drug screening due to its relevance in modeling the in vivo tumor tissue organization and microenvironment.2,3 The 3D multicellular spheroids could provide valuable information in a more timely and less expensive manner than directly proceeding from 2D cell culture experiments to animal (murine) models.
To facilitate the discovery of new therapeutics for NET patients, we have developed an in vitro 3D multicellular spheroids model using the human NET cell lines. The NET cells are plated in a non-adhesive agarose-coated 24-well plate and incubated under physiological conditions (5% CO2, 37 &deg;C) with a very slow agitation for 16-24 hr after plating. The cells form multicellular spheroids starting on the 3rd or 4th day. The spheroids become more spherical by the 6th day, at which point the drug treatments are initiated. The efficacy of the drug treatments on the NET spheroids is monitored based on the morphology, shape and size of the spheroids with a phase-contrast light microscope. The size of the spheroids is estimated automatically using a custom-developed MATLAB program based on an active contour algorithm. Further, we demonstrate a simple method to process the HistoGel embedding on these 3D spheroids, allowing the use of standard histological and immunohistochemical techniques. 
This is the first report on generating 3D spheroids using NET cell lines to examine the effect of therapeutic drugs. We have also performed histology on these 3D spheroids, and displayed an example of a single drug's effect on growth and proliferation of the NET spheroids. Our results support that the NET spheroids are valuable for further studies of NET biology and drug development.

We present a simple agarose overlay platform to grow 3D multicellular spheroids using neuroendocrine cancer cell lines. This method provides a very convenient way to examine the effect of therapeutic drugs on the neuroendocrine tumor cells. It could also help us establish human neuroendocrine tumor spheroids for cancer therapy.

인간 Neuroendocrine 종양 치료의 연구를위한 3 차원 모델로 인간 Neuroendocrine 종양 세포 라인

Neuroendocrine tumors (NETs) are rare tumors, with an incidence of two per 100,&#x200A;000 individuals per year, and they account for 0.5% of all human ...

A Murine Model of Stent Implantation in the Carotid Artery for the Study of Restenosis

Despite the considerable progress made in the stent development in the last decades, cardiovascular diseases remain the main cause of death in western countries. Beside the benefits offered by the development of different drug-eluting stents, the coronary revascularization bears also the life-threatening risks of in-stent thrombosis and restenosis. Research on new therapeutic strategies is impaired by the lack of appropriate methods to study stent implantation and restenosis processes. Here, we describe a rapid and accessible procedure of stent implantation in mouse carotid artery, which offers the possibility to study in a convenient way the molecular mechanisms of vessel remodeling and the effects of different drug coatings.

A model of stent implantation in mouse carotid artery is described. Compared to other similar methods, this procedure is very rapid, simple and accessible, offering the possibility to study in a convenient way the vascular wall reaction to different drug-eluting stents and the molecular mechanisms of restenosis.

재 협착의 연구에 대한 경동맥 스텐트 시술의 쥐과 모형

Despite  the considerable progress made in the stent development in the last decades,  cardiovascular diseases remain the main cause of death in western ...

Technical Aspects of the Mouse Aortocaval Fistula

Technical aspects of creating an arteriovenous fistula in the mouse are discussed. Under general anesthesia, an abdominal incision is made, and the aorta and inferior vena cava (IVC) are exposed. The proximal infrarenal aorta and the distal aorta are dissected for clamp placement and needle puncture, respectively. Special attention is paid to avoid dissection between the aorta and the IVC. After clamping the aorta, a 25 G needle is used to puncture both walls of the aorta into the IVC. The surrounding connective tissue is used for hemostatic compression. Successful creation of the AVF will show pulsatile arterial blood flow in the IVC. Further confirmation of successful AVF can be achieved by post-operative Doppler ultrasound.

The goal is to produce an arteriovenous fistula that is simple and reproducible. This method does not use sutures or glue adhesive. Therefore the samples can be used with the least amount of foreign materials for analysis.

마우스 Aortocaval 누관의 기술적 측면

Technical aspects of creating an arteriovenous fistula in the mouse are discussed. Under general anesthesia, an abdominal incision is made, and the aorta ...

A Murine Model of Subarachnoid Hemorrhage

In this video publication a standardized mouse model of subarachnoid hemorrhage (SAH) is presented. Bleeding is induced by endovascular Circle of Willis perforation (CWp) and proven by intracranial pressure (ICP) monitoring. Thereby a homogenous blood distribution in subarachnoid spaces surrounding the arterial circulation and cerebellar fissures is achieved. Animal physiology is maintained by intubation, mechanical ventilation, and continuous on-line monitoring of various physiological and cardiovascular parameters: body temperature, systemic blood pressure, heart rate, and hemoglobin saturation. Thereby the cerebral perfusion pressure can be tightly monitored resulting in a less variable volume of extravasated blood. This allows a better standardization of endovascular filament perforation in mice and makes the whole model highly reproducible. Thus it is readily available for pharmacological and pathophysiological studies in wild type and genetically altered mice.

A standardized mouse model of subarachnoid hemorrhage by intraluminal Circle of Willis perforation is described. Vessel perforation and subarachnoid bleeding are monitored by intracranial pressure monitoring. In addition various vital parameters are recorded and controlled to maintain physiologic conditions.

지주막 하 출혈의 생쥐 모델

In this video publication a standardized mouse model of subarachnoid  hemorrhage (SAH) is presented. Bleeding is induced by endovascular Circle of  Willis ...

A Murine Model of Arterial Restenosis: Technical Aspects of Femoral Wire Injury

Cardiovascular disease caused by atherosclerosis is the leading cause of death in the developed world. Narrowing of the vessel lumen, due to atherosclerotic plaque development or the rupturing of established plaques, interrupts normal blood flow leading to various morbidities such as myocardial infarction and stroke. In the clinic endovascular procedures such as angioplasty are commonly performed to reopen the lumen. However, these treatments inevitably damage the vessel wall as well as the vascular endothelium, triggering an excessive healing response and the development of a neointimal plaque that extends into the lumen causing vessel restenosis (re-narrowing). Restenosis remains a major cause of failure of endovascular treatments for atherosclerosis. Thus, preclinical animal models of restenosis are vitally important for investigating the pathophysiological mechanisms as well as translational approaches to vascular interventions. Among several murine experimental models, femoral artery wire injury is widely accepted as the most suitable for studies of post-angioplasty restenosis because it closely resembles the angioplasty procedure that injures both endothelium and vessel wall. However, many researchers have difficulty utilizing this model due to its high degree of technical difficulty. This is primarily because a metal wire needs to be inserted into the femoral artery, which is approximately three times thinner than the wire, to generate sufficient injury to induce prominent neointima. Here, we describe the essential surgical details to effectively overcome the major technical difficulties of this model. By following the presented procedures, performing the mouse femoral artery wire injury becomes easier. Once familiarized, the whole procedure can be completed within 20 min.

The mouse femoral artery wire injury model of restenosis is technically challenging. In this protocol we show the key technical details essential for successfully performing wire injury to induce consistent&#160;neointima for studies of restenosis.

동맥 재 협착의 쥐 모델 : 대퇴 와이어 부상의 기술적 측면

Cardiovascular disease caused by atherosclerosis is the leading cause of death in the developed world. Narrowing of the vessel lumen, due to ...

Isometric Contractility Measurement of the Mouse Mesenteric Artery Using Wire Myography

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and drugs. It is widely used in many studies on the physiological functions of vascular smooth muscle, the pathogenesis of vascular diseases such as hypertension, and the development of smooth muscle relaxant drugs. The mouse is a widely used model animal with a large pool of disease models and genetically modified strains. We introduced this method to measure the isometric contraction of mouse mesenteric artery in detail. A 1.4-mm segment of mouse mesenteric resistance artery was isolated and mounted on a myograph chamber by passing two steel wires through its lumen. After equilibration and normalization steps, the vessel segment was potentiated by a high-K+ solution twice prior to the contraction assay. As an example of the application of this method in drug development, we measured the relaxant effect of a novel natural substance, neoliensinine, isolated from a Chinese herb, embryos of the lotus seed (Nelumbo nucifera Gaertn.) on mouse mesenteric arteries. The vessel segments mounted on the myograph chamber were stimulated with a high-K+ solution. When the force tension reached a stable sustained phase, cumulative doses of neoliensinine were added to the chamber. We found that neoliensinine had a dose-dependent relaxant effect on smooth muscle contraction, thus suggesting that it bears potential activity against hypertension. In addition, as the vessel segment can survive at least 4 hours after mounting and maintain contractility induced by the high-K+ solution for many times, we suggest that the wire myograph system may be used for the time-consuming process of drug screening.

The wire myograph technique is used to investigate vascular smooth muscle functions and screen new drugs. We report a detailed protocol for measuring the isometric contractility of the mouse mesenteric artery and for screening new relaxants of vascular smooth muscle.

Myography 와이어를 사용 하 여 마우스 Mesenteric 동맥의 isometric 수축 측정

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and ...