Name: assessment of plasma coagulation on liver tissue in a large animal model in vivo
Uploaded: 2018-08-04T13:00:00.000+00:00
Duration: 6 min 23 s
Description: Watch this Scientific Journal Video about Assessment of Plasma Coagulation on Liver Tissue in a Large Animal Model In Vivo at JoVE.com

저자 아무 승인 있다.

동물 연구로 원리의 실험실 동물 관리 (건강의 국가 학회 간행물 출간: 8, 2011 년) 독일 입법에 의해 경 세 하는 규칙을 따라 했다. 공식 허가 정부 동물 관리 사무실 (Landesamt 위한 Natur, Umwelt und Verbraucherschutz 노르 트 베스트 팔 렌, Recklinghausen, 독일)에서 부여 됩니다.
1입니다. 동물
<ol><li>열린 감 금 소에 여성 독일 landrace 돼지 (25-30 kg 무게)를 사용 합니다.</li>
	<li>그룹 (아르곤과 헬륨) 당 5 동물을 사용 합니다.</li>
	<li>실험 하기 전에 적어도 1 주일에 대 한 환경에 순응을 동물 허용. 물에 대 한 무료 액세스와 함께 수술 이전 24 시간에 대 한 빠른 동물.</li>
</ol>2입니다. 마 취
<ol><li>Premedicate 동물 마 취 제 (15 mg/kg 몸 무게 [BW]), xylazine (10 mg/kg BW), 및 아트로핀 (0.1 mg/kg BW) 근육 주사와 마 취의 유도 하기 전에 10 분.</li>
	<li>주변 정 맥 접근 배치 22 게이지 정의 귀 정 맥에 의해 설정 됩니다.</li>
	<li>Propofol 2 mg/kg 체중의 i.v. 주입 하 여 전신 마 취를 유도.</li>
	<li>부정사 위치에 동물을 놓고 2 cm. 피하 조직의 무딘 준비를 통해 정 맥 찾기의 길이 jugular 홈에 경도 피부 절 개를 수행 합니다. 정, 다음 Seldinger 와이어를 삽입 합니다.</li>
	<li>정을 철회 하 고 가이드를 통해 14 백. 카 테 터를 삽입 합니다. 가이드 와이어를 철회. 카 테 터는 내선 번호로 연결 하 고 스트랩 또는 봉합 하 여 카 테 터를 흥분 시키는.</li>
	<li>바로 후 두 경 삽입를 혀 밖으로 당기고. 풀 다운은 후두개는 후 두 경의 팁을 사용 합니다. 성 대를 통해 튜브를 삽입 합니다. Glottis 아래 갑을 놓고 팽창 합니다.</li>
	<li>36와 42 mm Hg 사이 끝 갯벌 부분 탄소 이산화 긴장을 유지 하 20-26 호흡/min과 10 ml/kg의 해 일 볼륨에서 40% 산소로 환기.</li>
	<li>1-1.5%의 농도에서 isoflurane와 3-4 µ g/k g/h의 농도에 펜타닐 마 취를 유지 합니다.</li>
	<li>4 mL/kg의 초기 속도로 벨의 젖 산 솔루션 공급/h, 8 mL/kg의 일정 한 주입 속도에 개복 술 후 증가/h.</li>
</ol>3. 수술 및 플라즈마 응고
<ol><li>표준 수술 테이블에 부정사 위치에 동물을 배치 합니다.</li>
	<li>표준 외과 살 균 제를 적용 하 여 피부를 소독 (2-프로 판 올 45 g/100 g, 1-프로 판 올 10 g/100 g, 비페닐-2-ol 0.2 g/100 g) 3 번 수술 면봉으로.</li>
	<li>메스와 액을 넓은 중간 선 개복 술 칼 프로세스에서 수행 하 고 외과 견인 기를 설치.</li>
	<li>플라즈마 응고 장치에 스위치, 아르곤 또는 헬륨 가스 병 사용된 캐리어 가스에 따라 오픈. 원하는 대로 3 L/분 선택 응고 장치 출력 전력에 가스 흐름을 조정 합니다. 참고: 두 노블 가스, 아르곤 또는 헬륨, 플라즈마 응고에 대 한 사용할 수 있습니다. 응고 효과 비교할 수 있습니다. 자세한 내용은 참조7 를 참조 하십시오.</li>
	<li>앞에서 설명한7왼쪽된 간 엽에 플라즈마 응고를 수행 합니다. 티타늄 형 (사각 조리개 1 x 1 c m2)를 사용 하 여 응고 영역을 표준화. 5 응고 프로브 거리 1 cm의 s. 5 mm (그림 1)의 coagulations 사이의 짧은 거리와 다른 전원 설정을 coagulations는 수행할 수 있습니다.</li>
	<li>간의 수확에 대 한 모든 인 대 연결 간을 나눕니다. 분리 하 고 문맥 및 일반적인 담 관의 긴 부분을 떠나 우수한 십이지 장 굴곡 위 간 작은 꽃 자루를 나눕니다. 위와 간 아래 caval 정 맥을 분할 하 고 기관 검색.</li>
	<li>간, 수확 후 돼지 0.16 g/kg BW pentobarbital i.v. 관리에 의해 안락사 되었다.</li>
	<li>버스트 압력 실험에 대 한 날카로운가 위 왼쪽된 중간 간 엽의 절반을 resect. 플라즈마 응고 컷 (100W 출력) 표면 또는 섬유 소 실 란 트 (그림 2)와 함께 잘라 표면 밀봉.</li>
</ol>4. 미세 측정
참고: 레이저 도플러 분광학은 적혈구의 움직임으로 인 한 도플러 이동 측정을 통해 조직에 혈액 흐름을 확인할 수 있습니다. 레이저 신호 이동 적혈구의 수와 상관 한다. 레이저 도플러 분광학 임상 사용 (예: 이식 의학)과 여러 번15확인.
<ol><li>레이저 도플러 유량 계 및 분 광 광도 계에 전환 합니다. 플랫 프로브를 사용 합니다.</li>
	<li>흐름 및 속도 대 한 기준 측정을 가져가 라. 저장 하거나 값을 확인 합니다.</li>
	<li>3.5에서 설명 된 대로 응고를 수행 합니다.</li>
	<li>응고 사이트 및 측정 흐름 및 속도에 플랫 프로브를 배치 합니다. 다시 저장 하거나 값을 참고 합니다.</li>
	<li>Coagulator 장치의 모든 전원 설정에 대 한 반복 합니다.</li>
</ol>5. 온도 측정
<ol><li>(열감지 카메라, 노트북, 및 적외선 온도계)에 시스템을 전환 하 고 측정을 수행 하기 전에 적어도 1 시간 동안 실행 하자.</li>
	<li>초점을 조정 하 고 응고 사이트에 열감지 카메라에 프레임을 봅니다. 20 보다 더 큰 온도 해상도 1024 x 768 픽셀의 해상도와 적외선 시퀀스를 검출 될 수 있다 mK. 고려, 그 응고와 주변 조직의 지역-열 전달에 의해 영향을-보기 중간에 위치 하 고 있습니다. 참고: 그것은 가능한 최적 공간 해상도 대 한 프레임의 픽셀을 포함 해야 합니다.</li>
	<li>2 분 동안 열감지 카메라와 함께 간 표면에 플라즈마 coagulator과 응고 과정을 기록 합니다.</li>
	<li>온도 기록 분석 소프트웨어와 함께 이미지 시퀀스 분석: 관심의 영역을 정의. 참고: 소프트웨어 시간 해당 평균 온도 과정을 계산합니다.</li>
</ol>6. 응고 깊이 측정
<ol><li>날카로운가 위 왼쪽된 중간 간 엽 수확.</li>
	<li>1 cm 두께가 응고 사이트를 삭제할. 추가 처리를 위해 3 m m 두꺼운 경도 세그먼트를 잘라.</li>
	<li>밤새 중립 10%와 4 ° C에서 수정 조직 샘플 버퍼링 말린. 열 용융 점 이상 2 ° C 파라핀 하 고 분할 영역을 포함. 프로세스 하룻밤입니다.</li>
	<li>되며/오신 얼룩을 수행 합니다.<ol>
<li>Deparaffinize 및 이후 2에 담거 서 조직을 수 화 x 크 실 렌, 100% 에탄올 (EtOH), 95 %EtOH, 70 %EtOH, 이온된 H2O 2 분.</li>
		<li>3 분 위한 마이어의 되며 솔루션 조직 샘플을 얼룩.</li>
		<li>이제 5 분 동안 수돗물에 씻어.</li>
		<li>3 분 오신 솔루션 조직 얼룩.</li>
		<li>3 분 2 x EtOH 95% 그리고 크 실 렌에 린스. 표준 설치 매체와 함께 탑재.</li>
	</ol>
</li>
	<li>(현미경 연결 된 카메라, 이미징 소프트웨어)에 시스템을 전환 합니다. 40 X 확대와 함께 모든 섹션을 보기.</li>
	<li>40 X의 확대는 개체 마이크로미터의 이미지를 가져가 라. 재조정 목표 창에서 버튼을 누릅니다. 수동 보정을 선택 합니다. 100 µ m. 대화 상자에서 m m 입력 0, 1 마이크로미터 이미지에 라인을 그릴 하 고 확인을 누릅니다.</li>
	<li>보기에서 길이 선택 > 분석 컨트롤 > 주석 및 측정 창. 마우스로 응고 여백에 간 표면에서 측정 합니다. 내보내기 또는 결과 참고. 같은 슬라이드에 다른 위치에 측정을 반복 합니다. 참고: 응고 깊이 쉽게 정상적인 hepatocyte 코드와 수축 된 세포질, pyknotic 핵 및 출혈 영역 괴의 영역 사이의 날카로운 차이로 정상적인 간 조직에서 분화 될 수 있다.</li>
	<li>두 측정의 평균을 계산 합니다.</li>
</ol>7. 버스트 압력 측정
<ol><li>(자동 펌프, 압력 미터)에 시스템을 전환 합니다. 3.7 단계에 따라 간 견본을 준비 합니다. 참고: 3-방법-자 지를 통해 연결 된 두 개의 병렬 펌프를 사용 합니다. 1500 mm Hg의 최대 압력은 단일 펌프와 함께 얻을 수 없습니다.</li>
	<li>햅 틱 작은 꽃 자루가 위 문맥, 일반적인 간 동맥 및 담 즙 덕트를 격리 합니다. 문맥 overholt 집게와 클램프와 monofil 봉합 4-0와 선. 일반적인 간 동맥 overholt 집게를 죄 십시오 그리고 monofil 봉합 4-0와 선.</li>
	<li>일반적인 담 관으로 채널 16 카 테 터를 삽입 하 고 2-0 비단 봉합 사로 선. 자동 펌프 카 테 터를 연결, 압력 미터 (그림 3)와 함께 3 자 지를 설치 합니다.</li>
	<li>식 염 수로 관류 주사기를 채우십시오.</li>
	<li>99 mL/h의 배달 속도와 자동 펌프를 시작 합니다.</li>
	<li>버스트 압력 누설 및 기록에 대 한 잘라 간 표면과 압력 미터를 모니터링 합니다. 참고: 누설의 더 쉬운 인식을 위해 특허 블루는 식 염 수 (2 mL 특허 블루 + 18 mL 식 염 수)을 추가할 수 있습니다. 버스트 압력 압력 미터에 압력의 손실의 시간을 주의 하 여 관찰 하는 것이 쉽습니다.</li>
</ol>

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M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Adhesive+strength+of+fibrinogen-coated+collagen+patch+or+liquid+fibrin+sealant+in+an+experimental+liver+resection+model+in+pigs.">Adhesive strength of fibrinogen-coated collagen patch or liquid fibrin sealant in an experimental liver resection model in pigs.</a> Eur Surg Res Eur Chir Forsch Rech Chir Eur. 41 (3), 298-302 (2008).</li><li>Knobloch, K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Microcirculation+of+the+sternum+following+harvesting+of+the+left+internal+mammary+artery.">Microcirculation of the sternum following harvesting of the left internal mammary artery.</a> ThoracCardiovascSurg. 51, 255-259 (2003).</li><li>Kanzler, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Recommendation+for+severity+assessment+following+liver+resection+and+liver+transplantation+in+rats:+Part+I.">Recommendation for severity assessment following liver resection and liver transplantation in rats: Part I.</a> Lab Anim. 50 (6), 459-467 (2016).</li><li>Pehb&#246;ck, D., Dietrich, H., Klima, G., Paal, P., Lindner, K. H., Wenzel, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Anesthesia+in+swine+optimizing+a+laboratory+model+to+optimize+translational+research.">Anesthesia in swine optimizing a laboratory model to optimize translational research.</a> Anaesthesist. 64 (1), 65-70 (2015).</li><li>Nykonenko, A., V&#225;vra, P., Zon&#269;a, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Anatomic+Peculiarities+of+Pig+and+Human+Liver.">Anatomic Peculiarities of Pig and Human Liver.</a> Exp Clin Transplant Off J Middle East Soc Organ Transplant. 15 (1), 21-26 (2017).</li><li>Fechner, G., von Pezold, J., Luzar, O., Hauser, S., Tolba, R. H., M&#252;ller, S. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Modified+spectrometry+(O2C+device)+of+intraoperative+microperfusion+predicts+organ+function+after+kidney+transplantation:+a+pilot+study.">Modified spectrometry (O2C device) of intraoperative microperfusion predicts organ function after kidney transplantation: a pilot study.</a> Transplant Proc. 41 (9), 3575-3579 (2009).</li><li>Patterson, E. J., Scudamore, C. H., Owen, D. A., Nagy, A. G., Buczkowski, A. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Radiofrequency+ablation+of+porcine+liver+in+vivo:+effects+of+blood+flow+and+treatment+time+on+lesion+size.">Radiofrequency ablation of porcine liver in vivo: effects of blood flow and treatment time on lesion size.</a> AnnSurg. 227, 559-565 (1998).</li></ol>

저자는 공개 없다.

설치류 모델 간 수술에 대 한 오랜 시간16에 대 한 설정 됩니다. 그럼에도 불구 하 고, 큰 동물 모델 특정 이점을 제공:로 적용할 수 있는 인 간에 대 한 표준 수술 장비, 수술 기법 비교 임상 사용 표준 임상 평가 방법 수 없음 microsurgical 장비 필요 전송 실험. 예를 들어 표준 임상 혈액 검사 특별 한 실험실 테스트 방법 (그림 10)에 대 한 필요 없이 수행할 수 있...

아르곤 플라즈마 응고 (APC) 3 년간1,2보다 더 복 부 수술에서 널리 사용 되는 악기 이다. 그것은 이차 hemostasis의 성취에 대 한 표준 기술 간 씰링 하 여 주요 hepatectomy 잘라 나중 출혈3을 방지 하기 위해 표면. 플라즈마 응고는 이온화 가스의 아크를 통해 전기 에너지를 전달 하는 고주파 electrocautery의 특수 형태 이다. Monopolar electrothermal hemostasis, 제공이 비접촉 기술 조직4에 충실 하는 전극 방지의 이점이 있다. 이온화 가스 빔 자동으로 낮은 전기 저항 지역으로 이동 하 고 아직 포장해 다른 지역에 건조로 인해 저항 상승 때 멀리 설정 되어. 이 응고5,6의 균일 한 제한 된 깊이 생성합니다. 응고 효과 영향을 미치는 요소는 활성화 시간, 응고 장치 및 조직에는 프로브에서 거리의 전원 설정. 헬륨은 플라즈마 응고7사용할 수 있는 다른 캐리어 가스 이다. 최근 임상 연구에 집중된 임상 결과 보다는 조직학 및 기능적인 결과3,8,9, 체 외에서 조사10에 초점을 맞춘 실험 연구 하면서 또는 격리 끼얹는다 장기11에 실험.
기본 프로토콜 돼지에 인간 표준 장비를 사용 하 여 임상 응용에 가까운 큰 동물 모델에서 플라즈마 응고의 효과의 연구를 수 있습니다: 레이저 도플러 유량 계에 의해 미세 noninvasively 평가 및 분 광 광도 계,이 표시12,13표준 임상 도구입니다. 응고 하는 동안 온도 변화를 적외선 온도계와 열감지 카메라 모니터링 된다. 조직학 되며/오신의 조직 샘플의 수확 후 섹션 스테인드 응고의 깊이 측정 됩니다. 이차 hemostasis에 대 한 다른 방법 가진 비교에 대 한 파열 압력 실험 수행 됩니다. 앞에서 설명한 기법14, 달리 이러한 혈압 제외 explanted 장기에 실시 하는 효과 관련. 플라즈마 응고의 로컬 효과에 설명된 조사, 뿐만 아니라 돼지 모델에서 또한 표준 혈액 검사를 맡아 수 있다.

<table><tbody><tr><td>Xylazine 20 mg/mL</td><td>Vetoquinol GmbH</td><td>Xylapan</td><td></td></tr><tr><td>Ketamine 100 mg/mL</td><td>Ceva GmbH</td><td>Ceva Ketamine Injection</td><td></td></tr><tr><td>Atropine 100 mg / 10 mL</td><td>Dr. Franz K&amp;ouml;hler Chemie GmbH</td><td>Atropinsulfat K&amp;ouml;hler 100mg Amp.</td><td></td></tr><tr><td>Propofol</td><td>Fresenius Kabi GmbH</td><td>Propofol 1% MCT Fresenius</td><td></td></tr><tr><td>Fentanyl</td><td>KG Rotexmedica GmbH</td><td>Fentanyl 0,5mg Rotexmedica</td><td></td></tr><tr><td>Isoflurane</td><td>Abbot GmbH</td><td>Forene 100% (V/V) 250 mL</td><td></td></tr><tr><td>Ringer&#039;s lactate solution</td><td>Baxter Deutschland GmbH</td><td></td><td>sodium 131mmol/l, potassium 5 mmol/l, calcium 2 mmol/l, cloride 111 mmol/l, lactate 29 mmol/l</td></tr><tr><td>Surgical disinfactant</td><td>Sch&amp;uuml;lke &amp;amp; Mayr GmbH</td><td>Kodan Tinktur forte gef&amp;auml;rbt 1l 104804</td><td></td></tr><tr><td>Motorized microscope</td><td>Nikon Instruments Europe</td><td>Eclipse TE2000-E</td><td></td></tr><tr><td>Microscope camera</td><td>Nikon Instruments Europe</td><td>Digitalsight DS-Qi1Mc</td><td></td></tr><tr><td>Imaging software</td><td>Nikon Instruments Europe</td><td>NIS elements Vers. 4.40</td><td></td></tr><tr><td>Plasma coagulator</td><td>S&amp;ouml;ring GmbH</td><td>CPC-1000</td><td></td></tr><tr><td>Argon gas</td><td>Linde AG</td><td>Argon 4.8&amp;nbsp;</td><td></td></tr><tr><td>Helium gas</td><td>Linde AG</td><td>Helium 4.8</td><td></td></tr><tr><td>O2C</td><td>LEA Medizintechnik GmbH</td><td>O2C Version 1212</td><td>with LF-2 or LF-3 probe</td></tr><tr><td>Infrared thermometer</td><td>Voltcraft</td><td>VOLTCRAFT IR 260-8S</td><td></td></tr><tr><td>Thermographic camera</td><td>InfraTec GmbH</td><td>VarioCAM HD head 820</td><td></td></tr><tr><td>Thermographic analysis software</td><td>InfraTec GmbH</td><td>IRBIS 3</td><td></td></tr><tr><td>Mayer&#039;s Hematoxylin solution</td><td></td><td>Merck 1.09249</td><td></td></tr><tr><td>Eosin solution</td><td>VWR International GmbH</td><td>Merck 1.09844</td><td></td></tr><tr><td>Rollerpump Masterflex L/S easy Load</td><td>Cole-Parmer Instrument Company</td><td>model 7518-10</td><td></td></tr><tr><td>Perfusorpump</td><td>B. Braun Melsungen AG</td><td>Perfusor secura FT</td><td></td></tr><tr><td>Digital pressure meter</td><td>Greisinger electronic</td><td>GMH 3161</td><td></td></tr><tr><td>Perfusorsyringe, 50 mL</td><td>B. Braun Melsungen AG</td><td>REF 8728810 F</td><td></td></tr><tr><td>Perfusor line, Type IV Standard, PVC Luer lock</td><td>B. Braun Melsungen AG</td><td>REF 8722960</td><td></td></tr><tr><td>3-Way stopcock, Dicofix C35C</td><td>B. Braun Melsungen AG</td><td>REF 16494 C</td><td></td></tr><tr><td>Silk 2-0. 3 metric</td><td>Resorba</td><td>REF H5F</td><td></td></tr><tr><td>Vicryl 4-0 Sutupak</td><td>Ethicon</td><td>V1224H</td><td></td></tr><tr><td>NaCl 0.9 %</td><td>B. Braun Melsungen AG</td><td></td><td></td></tr></tbody></table>

assessment of plasma coagulation on liver tissue in a large animal model in vivo

플라즈마 응고 electrocautery의 형태로 나중 단계에서 출혈을 방지 하기 위해 주요 hepatectomy 후 큰 간 잘라 표면 밀봉 수십 년 간 수술에 사용 됩니다. 간 조직에 플라즈마 응고의 정확한 효과 가난 하 게 검사 합니다. 우리의 돼지 모델에서 응고 효과 임상 응용 프로그램에 가까운 시험 될 수 있다. 결합 된 레이저 도플러 유량 계 및 분 광 광도 계 문서 미세 변경 8 m m 조직 깊이에서 응고 하는 동안 noninvasively, 주관적인 임상 인상 넘어 hemostasis에 대 한 정량 정보를 제공 합니다. 응고 사이트에서 온도 응고 하는 동안 적외선 온도계 사전 및 사후 응고와 열감지 카메라 평가, 가스 빔 온도 측정 소자의 상한값 때문에 가능 하지 않습니다. 응고의 깊이 되며/오신 교정 후 개체 마이크로미터 섹션을 물 들일에 현미경으로 측정 되 고 전원 설정-응고 깊이 관계에 대 한 정확한 정보를 제공. 그것은 큰 혈관을 밀봉 하기 위하여 플라즈마 coagulator 불가능으로 씰링 효과 담 관에 시험 된다. 버스트 압력 실험 수행 됩니다 밖으로 배제 혈압 explanted 장기 효과 관련.

미세: 플라즈마 응고 다음 hemostasis 위한 진단 장치를 활용 하 여 미세 혈관 변화에 의해 설명할 수 있습니다. 142.7 ± 76.08의 기준 값에서 모 세관 혈액 흐름 (임의의 단위 (AU)로 표시) 감소 57.78 ± 49.57 하는 누구나 48.5 ± 7.26에 25 W 장치 출력 전력, AU AU 50 W와 5.04 ± 1.31 AU 100 W (그림 4).
온도:</...

Watch this Scientific Journal Video about Assessment of Plasma Coagulation on Liver Tissue in a Large Animal Model In Vivo at JoVE.com

Assessment of Plasma Coagulation on Liver Tissue in a Large Animal Model In Vivo

여기 선물이 비보에간 조직에 플라즈마 응고를 실험적으로 평가 하는 프로토콜. 돼지 모델에서 미세 레이저 도플러 검사, 응고 깊이 조직학 측정, 온도 적외선 온도계와 열감지 카메라와 덕트 씰링 효과 의해 응고 사이트에서 버스트 압력에 의해 문서화 실험입니다.

플라즈마 응고는 큰 동물 모델에서 에서 Vivo에서 간 조직에의 평가

Plasma coagulation as a form of electrocautery is used in liver surgery for decades to seal the large liver cut surface after major hepatectomy to prevent ...

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Medicine

Assessment of Plasma Coagulation on Liver Tissue in a Large Animal Model In Vivo

8.8K 조회수.  University of Bonn. 여기 선물이 비보에간 조직에 플라즈마 응고를 실험적으로 평가 하는 프로토콜. 돼지 모델에서 미세 레이저 도플러 검사, 응고 깊이 조직학 측정, 온도 적외선 온도계와 열감지 카메라와 덕트 씰링 효과 의해 응고 사이트에서 버스트 압력에 의해 문서화 실험입니다.

비디오: Assessment of Plasma Coagulation on Liver Tissue in a Large Animal Model In Vivo

A Novel Surgical Technique As a Foundation for In Vivo Partial Liver Engineering in Rat

Organ engineering is a novel strategy to generate liver organ substitutes that can potentially be used in transplantation. Recently, in vivo liver engineering, including in vivo organ decellularization followed by repopulation, has emerged as a promising approach over ex vivo liver engineering. However, postoperative survival was not achieved. The aim of this study is to develop a novel surgical technique of in vivo selective liver lobe perfusion in rats as a prerequisite for in vivo liver engineering. We generate a circuit bypass only through the left lateral lobe. Then, the left lateral lobe is perfused with heparinized saline. The experiment is performed with 4 groups (n = 3 rats per group) based on different perfusion times of 20 min, 2 h, 3 h, and 4 h. Survival, as well as the macroscopically visible change of color and the histologically determined absence of blood cells in the portal triad and the sinusoids, is taken as an indicator for a successful model establishment. After selective perfusion of the left lateral lobe, we observe that the left lateral lobe, indeed, turned from red to faint yellow. In a histological assessment, no blood cells are visible in the branch of the portal vein, the central vein, and the sinusoids. The left lateral lobe turns red after reopening the blocked vessels. 12/12 rats survived the procedure for more than one week. We are the first to report a surgical model for in vivo single liver lobe perfusion with a long survival period of more than one week. In contrast to the previously published report, the most important advantage of the technique presented here is that perfusion of 70% of the liver is maintained throughout the whole procedure. The establishment of this technique provides a foundation for in vivo partial liver engineering in rats, including decellularization and recellularization.

A Novel Surgical Technique As a Foundation for In Vivo Partial Liver Engineering in Rat

We establish a novel surgical technique for an in vivo single liver lobe perfusion model in rat as a prerequisite for further studying in vivo partial liver engineering in the future.

Vivo에서 부분 간 엔지니어링을에서 위한 쥐를 기초로 새로운 수술 기법

Organ engineering is a novel strategy to generate liver organ substitutes that can potentially be used in transplantation. Recently, in vivo liver ...

연구

생체공학

Uncontrolled Hemorrhagic Shock Modeled via Liver Laceration in Mice with Real Time Hemodynamic Monitoring

Uncontrolled hemorrhage is an important cause of preventable deaths among trauma patients. We have developed a murine model of uncontrolled hemorrhage via a liver laceration that results in consistent blood loss, hemodynamic alterations, and survival.
Mice undergo a standardized resection of the left-middle lobe of the liver. They are allowed to bleed without mechanical intervention. Hemostatic agents can be administered as pre-treatment or rescue therapy depending on the interest of the investigator. During the time of hemorrhage, real-time hemodynamic monitoring via a left femoral arterial line is performed. Mice are then sacrificed, blood loss is quantified, blood is collected for further analysis, and organs are harvested for analysis of injury. Experimental design is described to allow for simultaneous testing of multiple animals.
Liver hemorrhage as a model of uncontrolled hemorrhage exists in the literature, primarily in rat and porcine models. Some of these models utilize hemodynamic monitoring or quantify blood loss but lack consistency. The present model incorporates quantification of blood loss, real-time hemodynamic monitoring in a murine model that offers the advantage of using transgenic lines and a high-throughput mechanism to further investigate the pathophysiologic mechanisms in uncontrolled hemorrhage.

Uncontrolled hemorrhage, an important cause of mortality among trauma patients, can be modeled using a standard liver laceration in a murine model. This model results in consistent blood loss, survival, and allows for testing hemostatic agents. This article provides the step-by-step process to perform this valuable model.

실시간 혈류 역학 모니터링을 이용한 마우스의 간경화로 모델링 된 통제 할 수없는 출혈성 쇼크

Uncontrolled hemorrhage is an important cause of preventable deaths among trauma patients. We have developed a murine model of uncontrolled hemorrhage via ...

의학

Rat Model of the Associating Liver Partition and Portal Vein Ligation for Staged Hepatectomy (ALPPS) Procedure

Recent clinical data support an aggressive surgical approach to both primary and metastatic liver tumors. For some indications, like colorectal liver metastases, the amount of liver tissue left behind after liver resection has become the main limiting factor of resectability of large or multiple liver tumors. A minimal amount of functional tissue is required to avoid the severe complication of post-hepatectomy liver failure, which has high morbidity and mortality. Inducing liver growth of the prospective remnant prior to resection has become more established in liver surgery, either in the form of portal vein embolization by interventional radiologists or in the form of portal vein ligation several weeks prior to resection. Recently, it was shown that liver regeneration is more extensive and rapid, when the parenchymal transection is added to portal vein ligation in a first stage and then, after only one week of waiting, resection performed in a second stage (Associating Liver Partition and Portal vein ligation for Staged hepatectomy = ALPPS). ALPPS has rapidly become popular across the world, but has been criticized for its high perioperative mortality. The mechanism of accelerated and extensive growth induced by this procedure has not been well understood. Animal models have been developed to explore both the physiological and molecular mechanisms of accelerated liver regeneration in ALPPS. This protocol presents a rat model that allows mechanistic exploration of accelerated regeneration.

Rat Model of the Associating Liver Partition and Portal Vein Ligation for Staged Hepatectomy ALPPS Procedure

Inducing rapid liver hypertrophy using Associating Liver Partition and Portal vein ligation for a Staged hepatectomy (ALPPS) has been proposed for resection of borderline resectable liver tumors. This model may elucidate mechanisms involved in rapid hypertrophy and allows testing of drugs that promote or block the acceleration of regeneration.

연결 간 파티션과 무대 Hepatectomy (ALPPS) 절차에 대 한 포털 정 맥 결 찰의 쥐 모델

Recent clinical data support an aggressive surgical approach to both primary and metastatic liver tumors. For some indications, like colorectal liver ...

Invasive Hemodynamic Characterization of the Portal-hypertensive Syndrome in Cirrhotic Rats

This is a detailed protocol describing invasive hemodynamic measurements in cirrhotic rats for the characterization of portal hypertensive syndrome. Portal hypertension (PHT) due to cirrhosis is responsible for the most severe complications in patients with liver disease. The full picture of the portal hypertensive syndrome is characterized by increased portal pressure (PP) due to the increased intrahepatic vascular resistance (IHVR), hyperdynamic circulation, and increased splanchnic blood flow. Progressive splanchnic arterial vasodilation and increased cardiac output with elevated heart rate (HR) but low arterial pressure characterizes the portal hypertensive syndrome.
Novel therapies are currently being developed that aim to decrease PP by either targeting IHVR or increased splanchnic blood flow &#8212; but side effects on systemic hemodynamics may occur. Thus, a detailed characterization of portal venous, splanchnic, and systemic hemodynamic parameters, including measurement of PP, portal venous blood flow (PVBF), mesenteric arterial blood flow, mean arterial pressure (MAP), and HR is needed for preclinical evaluation of the efficacy of novel treatments for PHT. Our video article provides the reader with a structured protocol for performing invasive hemodynamic measurements in cirrhotic rats. In particular, we describe the catheterization of the femoral artery and the portal vein via an ileocolic vein and the measurement of portal venous and splanchnic blood flow via perivascular Doppler-ultrasound flow probes. Representative results of different rat models of PHT are shown.

Here we describe a detailed protocol for invasive measurements of hemodynamic parameters including portal pressure, splanchnic blood flow, and systemic hemodynamics in order to characterize the portal hypertensive syndrome in rats.

포털-고혈압 증후군 Cirrhotic 쥐에서의 침입 Hemodynamic 특성화

This is a detailed protocol describing invasive hemodynamic measurements in cirrhotic rats for the characterization of portal hypertensive syndrome. ...

A Small Animal Model of Ex Vivo Normothermic Liver Perfusion

There is a significant shortage of liver allografts available for transplantation, and in response the donor criteria have been expanded. As a result, normothermic ex vivo liver perfusion (NEVLP) has been introduced as a method to evaluate and modify organ function. NEVLP has many advantages in comparison to hypothermic and subnormothermic perfusion including reduced preservation injury, restoration of normal organ function under physiologic conditions, assessment of organ performance, and as a platform for organ repair, remodeling, and modification. Both murine and porcine NEVLP models have been described. We demonstrate a rat model of NEVLP and use this model to show one of its important applications &#8212; the use of a therapeutic molecule added to liver perfusate. Catalase is an endogenous reactive oxygen species (ROS) scavenger and has been demonstrated to decrease ischemia-reperfusion in the eye, brain, and lung. Pegylation has been shown to target catalase to the endothelium. Here, we added pegylated-catalase (PEG-CAT) to the base perfusate and demonstrated its ability to mitigate liver preservation injury. An advantage of our rodent NEVLP model is that it is inexpensive in comparison to larger animal models. A limitation of this study is that it does not currently include post-perfusion liver transplantation. Therefore, prediction of the function of the organ post-transplantation cannot be made with certainty. However, the rat liver transplant model is well established and certainly could be used in conjunction with this model. In conclusion, we have demonstrated an inexpensive, simple, easily replicable NEVLP model using rats. Applications of this model can include testing novel perfusates and perfusate additives, testing software designed for organ evaluation, and experiments designed to repair organs.

A Small Animal Model of Ex Vivo Normothermic Liver Perfusion

There is a significant liver donor shortage, and criteria for liver donors have been expanded. Normothermic ex vivo liver perfusion (NEVLP) has been developed to evaluate and modify organ function. This study demonstrates a rat model of NEVLP and tests the ability of pegylated-catalase, to mitigate liver preservation injury.

Ex Vivo Normothermic 간 관류의 작은 동물 모델

There is a significant shortage of liver allografts available for transplantation, and in response the donor criteria have been expanded. As a result, ...

Ferric Chloride-induced Canine Carotid Artery Thrombosis: A Large Animal Model of Vascular Injury

Occlusive arterial thrombosis leading to cerebral ischemic stroke and myocardial infarction contributes to ~13 million deaths every year globally. Here, we have translated a vascular injury model from a small animal into a large animal (canine), with slight modifications that can be used for pre-clinical screening of prophylactic and thrombolytic agents. In addition to the surgical methods, the modified protocol describes the step-by-step methods to assess carotid artery canalization by angiography, detailed instructions to process both the brain and carotid artery for histological analysis to verify carotid canalization and cerebral hemorrhage, and specific parameters to complete an assessment of downstream thromboembolic events by utilizing magnetic resonance imaging (MRI). In addition, specific procedural changes from the previously well-established small animal model necessary to translate into a large animal (canine) vascular injury are discussed.

Here, we present the modifications necessary to a well characterized and commonly used small animal ferric chloride-induced (FeCl3) carotid artery injury model for use in a large animal vascular injury model. The resulting model can be utilized for pre-clinical trial assessment of both prophylactic and thrombolytic pharmacological and mechanical interventions.

제 2 철 염화 유도 개 경 동맥 혈전 증: 혈관 손상의 큰 동물 모델

Occlusive arterial thrombosis leading to cerebral ischemic stroke and myocardial infarction contributes to ~13 million deaths every year globally. Here, ...

Murine Precision-Cut Liver Slices as an Ex Vivo Model of Liver Biology

Understanding the mechanisms of liver injury, hepatic fibrosis, and cirrhosis that underlie chronic liver diseases (i.e., viral hepatitis, non-alcoholic fatty liver disease, metabolic liver disease, and liver cancer) requires experimental manipulation of animal models and in vitro cell cultures. Both techniques have limitations, such as the requirement of large numbers of animals for in vivo manipulation. However, in vitro cell cultures do not reproduce the structure and function of the multicellular hepatic environment. The use of precision-cut liver slices is a technique in which uniform slices of viable mouse liver are maintained in laboratory tissue culture for experimental manipulation. This technique occupies an experimental niche that exists between animal studies and in vitro cell culture methods. The presented protocol describes a straightforward and reliable method to isolate and culture precision-cut liver slices from mice. As an application of this technique, ex vivo liver slices are treated with bile acids to simulate cholestatic liver injury and ultimately assess the mechanisms of hepatic fibrogenesis.

This protocol provides a simple and reliable method for the production of viable precision-cut liver slices from mice. The ex vivo tissue samples can be maintained under laboratory tissue culture conditions for multiple days, providing a flexible model to examine liver pathobiology.

뮤린 정밀 절단 간 슬라이스 간 생물학의 Ex Vivo 모델로

Understanding the mechanisms of liver injury, hepatic fibrosis, and cirrhosis that underlie chronic liver diseases (i.e., viral hepatitis, non-alcoholic ...

Porcine Liver Transplantation Without Veno-Venous Bypass As an Extended Criteria Donor Model

Liver transplantation is regarded as the gold standard for the treatment of a variety of fatal hepatic diseases. However, unsolved issues of chronic graft failure, ongoing organ donor shortages, and the increased use of marginal grafts call for the improvement of current concepts, such as the implementation of organ machine perfusion. In order to evaluate new methods of graft reconditioning and modulation, translational models are required. With respect to anatomical and physiological similarities to humans and recent progress in the field of xenotransplantation, pigs have become the main large animal species used in transplantation models. After the initial introduction of a porcine orthotopic liver transplant model by Garnier et al. in 1965, several modifications have been published over the past 60 years.
Due to specifies-specific anatomical traits, a veno-venous bypass during the anhepatic phase is regarded as a necessity to reduce intestinal congestion and ischemia resulting in hemodynamic instability and perioperative mortality. However, the implementation of a bypass increases the technical and logistical complexity of the procedure. Furthermore, associated complications such as air embolism, hemorrhage, and the need for a simultaneous splenectomy have been reported previously.
In this protocol, we describe a model of porcine orthotopic liver transplantation without the use of a veno-venous bypass. The engraftment of donor livers after static cold storage of 20 h - simulating extended criteria donor conditions - demonstrates that this simplified approach can be performed without significant hemodynamic alterations or intraoperative mortality and with regular uptake of liver function (as defined by bile production and liver-specific CYP1A2 metabolism). The success of this approach is ensured by an optimized surgical technique and a sophisticated anesthesiologic volume and vasopressor management.
This model should be of special interest for workgroups focusing on the immediate postoperative course, ischemia-reperfusion injury, associated immunological mechanisms, and the reconditioning of extended criteria donor organs.

In this protocol, a model of porcine orthotopic liver transplantation after static cold storage of donor organs for 20 h without the use of a veno-venous bypass during engraftment is described. The approach uses a simplified surgical technique with minimization of the anhepatic phase and sophisticated volume and vasopressor management.

확장 기준 기증자 모델로서의 정맥 우회술이 없는 돼지 간 이식

Liver transplantation is regarded as the gold standard for the treatment of a variety of fatal hepatic diseases. However, unsolved issues of chronic graft ...

Tail Vein Transection Bleeding Model in Fully Anesthetized Hemophilia A Mice

Tail bleeding models are important tools in hemophilia research, specifically for the assessment of procoagulant effects. The tail vein transection (TVT) survival model has been preferred in many settings due to sensitivity to clinically relevant doses of FVIII, whereas other established models, such as the tail clip model, require higher levels of procoagulant compounds. To avoid using survival as an endpoint, we developed a TVT model establishing blood loss and bleeding time as endpoints and full anesthesia during the entire experiment. Briefly, anesthetized mice are positioned with the tail submerged in temperate saline (37&#176;C) and dosed with the test compound in the right lateral tail vein. After 5 min, the left lateral tail vein is transected using a template guide, the tail is returned to the saline, and all bleeding episodes are monitored and recorded for 40 min while collecting the blood. If no bleeding occurs at 10 min, 20 min, or 30 min post-injury, the clot is challenged gently by wiping the cut twice with a wet gauze swab. After 40 min, blood loss is quantified by the amount of hemoglobin bled into the saline. This fast and relatively simple procedure results in consistent and reproducible bleeds. Compared to the TVT survival model, it uses a more humane procedure without compromising sensitivity to pharmacological intervention. Furthermore, it is possible to use both genders, reducing the total number of animals that need to be bred, in adherence with the principles of 3R&#39;s. A potential limitation in bleeding models is the stochastic nature of hemostasis, which can reduce the reproducibility of the model. To counter this, manual clot disruption ensures that the clot is challenged during monitoring, preventing primary (platelet) hemostasis from stopping bleeding. This addition to the catalog of bleeding injury models provides an option to characterize procoagulant effects in a standardized and humane manner.

The refined tail vein transection (TVT) bleeding model in anesthetized mice is a sensitive in vivo method for the assessment of hemophilic bleeding. This optimized TVT bleeding model uses blood loss and bleeding time as endpoints, refining other models and avoiding death as an endpoint.

꼬리 정맥 transection 출혈 모델 완전 마취 혈우병 A 마우스에서

Tail bleeding models are important tools in hemophilia research, specifically for the assessment of procoagulant effects. The tail vein transection (TVT) ...

생물학

Intravital Microscopy to Study Platelet-Leukocyte-Endothelial Interactions in the Mouse Liver

Inflammation and thrombosis are complex processes that occur primarily in the microcirculation. Although standard histology may provide insight into the end pathway for both inflammation and thrombosis, it is not capable of showing the temporal changes that occur throughout the time course of these processes. Intravital microscopy (IVM) is the use of live-animal imaging to gain temporal insight into physiologic processes in vivo. This method is particularly powerful when assessing cellular and protein interactions within the circulation due to the rapid and sequential events that are often necessary for these interactions to occur. While IVM is an extremely powerful imaging methodology capable of viewing complex processes in vivo, there are a number of methodological factors that are important to consider when planning an IVM study. This paper outlines the process of conducting intravital imaging of the liver, identifying important considerations and potential pitfalls that may arise. Thus, this paper describes the use of IVM to study platelet-leukocyte-endothelial interactions in liver sinusoids to study the relative contributions of each in different models of acute liver injury.

Intravital microscopy is a powerful tool that provides insight into both the temporal and spatial relationships of rapid and/or sequential processes. Herein, we describe a protocol to assess both protein-protein interactions and platelet-neutrophil-endothelial interactions in liver sinusoids in a murine model of experimental sepsis (endotoxemia).

Inflammation and thrombosis are complex processes that occur primarily in the microcirculation. Although standard histology may provide insight into the ...