Este estudo foi financiado pela &quot;Deutsche Forschungsgemeinschaft&quot; (DFG). K. Schleimer foi apoiado pelo Lise Meitner Sociedade do Ministério da Ciência e Pesquisa da NRW. Gostaríamos de agradecer a senhora Mary-Joan Blümich pelo seu excelente assistência edição. Dr. Maria Kokozidou foi financiada a Fresenius 2011_A61.

1. Animais Experimentais <ol><li> Obter uma aprovação do Comitê de Ética local para todos os experimentos que serão realizados, de acordo com as diretrizes de proteção animal FELASA (K 40, 17/99). </li><li> Use ratos Lewis machos disponíveis para compra a partir de qualquer criador comercial (por exemplo, Charles River Wiga GmbH, Sulzfeld, Alemanha). Aqui, 224 animais (112 doadores, 112 destinatários) foram utilizados para realizar 112 transplantes. Os receptores foram sacrificados seis semanas de pós-operatório. No momento da operação, os animais devem pesar 250-350 g. </li><li> Manter os animais em quartos climatizados com acesso gratuito à comida e água. </li></ol> 2 Anestesia e preparação operativo <ol><li> Colocar o rato dentro de uma caixa de plexiglas ligado à máquina de anestesia com uma taxa de fluxo de oxigénio de 1 L / min, e um vaporizador de isofluorano com 5% de gás anestésico para induzir a anestesia. Manter a anestesia em 1,5% isofluorano. </li><li> Colocar os ratos emdecúbito dorsal sobre uma placa de aquecimento. </li><li> Retire a pele do abdômen com um creme de depilação. </li><li> Prepare um campo cirúrgico estéril da seguinte forma: Use um desinfetante para limpar a bancada. Para a desinfecção de clorexidina a utilização local de incisão primeira, segunda e álcool, finalmente, uma solução de esfrega (por exemplo, cloro-hexidina e 70% de álcool isopropílico). Usar um vestido cirúrgico descartável e luvas cirúrgicas. </li><li> Use instrumentos de microcirurgia estéreis. </li><li> Use um microscópio de operação em ampliação de 12-20 vezes. </li></ol> 3 Procedimento Cirúrgico <ol><li> Operação do Doador Preparação <ol><li> Realize uma laparotomia mediana e ressecção do lobo lateral esquerdo eo lobo médio do enxerto, como descrito por Higgins. 14 </li><li> Em seguida, preparar o ligamento hepatoduodenal. Primeiro ligadura do ducto biliar, a fim de induzir a congestão e dilatação do ducto biliar. Em seguida, ligar e transecto a gastroduodenal veia e artéria. Em seguida, isolar o tronco celíaco e da artéria hepática comum e transecto da artéria gástrica esquerda e artéria esplênica. Inciso do ducto biliar congestionado, e introduzir um stent 20-G para ele. O posicionamento do stent tem de assegurar a drenagem de todo o sistema de ductos biliares hilares. Transecto do ducto biliar distal ao stent. </li><li> Isolar o segmento infrahepatic da veia cava logo acima da veia renal direita. Perfusão </li><li> Após ligadura da aorta acima do tronco celíaco, fixar a veia porta e inciso lo. Introduzir o cateter de perfusão (14 L) para dentro da veia portal, e efectuar a perfusão com 30 ml de triptofano--cetoglutarato de histidina (HTK) -solution (12 cm de H2O, 4 ° C). </li><li> Cortar o diafragma e a veia cava supra-hepática para permitir que o perfusato para fluir para fora. </li><li> Extirpar o tronco celíaco, juntamente com um remendo da aorta e lave-o com uma solução de histidina-triptofano-cetoglutarato. Explantação </li><li> Transecto a veia cava infrahepatic logo acima da veia renal direita. </li><li> Em seguida, coloque um stent com um diâmetro de 0,3 milímetros na veia porta e corrigi-lo com uma ligadura. Durante a operação seguinte, na veia porta é arterializada através da artéria renal direita. Transecto a veia portal distal ao stent. </li><li> Após ligadura da veia cava supra-hepática, transecto-lo juntamente com um manguito de diafragma. </li><li> Explante enxerto e armazená-lo em histidina-triptofano-cetoglutarato (HTK) -solution a 4 ° Celsius em um saco plástico com gelo. Suturas lugar de canto (8-0 Ethilon) na veia cava infrahepatic. </li></ol></li><li> Operação Destinatário <ol><li> Realize uma laparotomia mediana e preparar a veia cava infrahepatic. Em seguida, isolar a artéria renal direita e do ureter, ligar o ureter e transecção ele. Fixar a artéria renal direita em sua origem, em seguida, ligar a veia renal direita. Em seguida, após a transecção da re tantovasos Nal, realizar a nefrectomia. Posteriormente, encurtar a artéria renal direita, lave-o com solução de heparina salina (soro fisiológico 40 UI de heparina / ml). </li><li> Transecto a veia renal direita proximal à ligadura após o pinçamento proximal e distal da veia cava infrahepatic para criar um lúmen ovais na veia cava infrahepatic. Lavar a veia cava com uma solução salina com heparina. Implantar o enxerto na região renal direita do abdômen do destinatário. </li><li> Anastomosar infrahepatic fim-lateral da veia cava do doador para a veia cava recipient`s com sutura contínua 8-0 Ethilon. </li><li> Após lavagem com solução de heparina / solução salina, introdução do stent na veia portal para a artéria renal direita (Figura 3). Ligue a veia porta ea artéria renal com dois pontos (8-0 Ethilon). Fixe o stent ligando na artéria renal com uma ligadura (trançado de seda preta 7-0). </li><li> Para permitir a reperfusão, primeiro retire a braçadeira arterial, em seguida, ambosbraçadeiras venoso (Figura 4). </li><li> Prepare a aorta infra-renal, em seguida, coloque uma proximal e distal e inciso microclampe a aorta direita entre os dois grampos. Anastomose término-lateral entre a aorta e tronco celíaco do doador, usando um tapa-aórtica, com sutura contínua 10-0 Ethilon. </li><li> Inserir o canal biliar para o duodeno: Aplicar uma sutura em bolsa na parede do duodeno, fazer uma pequena incisão na parede do duodeno à direita no meio do em bolsa. Introduzir o ducto biliar stent para o duodeno, e aperte o sutura em bolsa. Finalmente, segurar o duodeno e do ducto biliar firmemente junto com dois pontos (8-0 Ethilon) (Figura 5). </li><li> Em seguida, realizar uma ressecção do fígado nativo, primeiro descrito por Emond et al. para imitar insuficiência hepática aguda. sete </li><li> Fixar a veia cava supra-hepática do enxerto, em conjunto com um rotador de diafragma, a parede abdominal lateral do recipient com Vicryl 4-0 sutura. Lavar o abdômen com solução salina (Figura 6). </li><li> Para a camada muscular utilizar um fio de sutura 4-0 Vicryl execução e para a sutura contínua cutânea 3-0 Vicryl. </li></ol></li></ol> 4 Recuperação e manejo pós-operatório <ol><li> Deixe os animais recuperar da anestesia sob uma lâmpada de aquecimento infravermelho. </li><li> Injectar buprenorfina a uma dose de 0,03-0,05 mg / kg de peso corporal todas as 8-12 horas sc durante 2 dias. </li><li> Examine os animais relativas ao estado geral e sinais de doença ou infecções. </li></ol> 5 Cálculo do fígado inicial Peso Native Calcula-se o peso inicial de fígado nativo da seguinte maneira: subtrair o peso dos lóbulos do fígado ressecados a partir do peso do fígado completo (2,66% do peso corporal (este valor, com um desvio padrão de 0,49% +/- foi determinada em experiências anteriores com n = 80 animais da mesma estirpe e umage)) 15.

<ol>
	<li>Boudjema, 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=Auxiliary+liver+transplantation+for+fulminant+and+subfulminant+hepatic+failure.">Auxiliary liver transplantation for fulminant and subfulminant hepatic failure.</a> Transplantation. 59, 218-223 (1995).</li><li>Boudjema, K., Jaeck, D., Sim&#233;oni, U., Bientz, J., Chenard, M. P., Brunot, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Temporary+auxiliary+liver+transplantation+for+subacute+liver+failure+in+a+child.">Temporary auxiliary liver transplantation for subacute liver failure in a child.</a> The Lancet. 342, 778-779 (1993).</li><li>Chenard-Neu, M. P., 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=Auxiliary+liver+transplantation:+Regeneration+of+the+native+liver+and+outcome+in+30+patients+with+fulminant+hepatic+failure+-+a+multicenter+European+study.">Auxiliary liver transplantation: Regeneration of the native liver and outcome in 30 patients with fulminant hepatic failure - a multicenter European study.</a> Hepatology. 23, 1119-1127 (1996).</li><li>Gubernatis, G., Pichlmayr, R., Kemnitz, J., Gratz, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Auxiliary+partial+orthotopic+liver&#8211;transplantation+(APOLT)+for+fulminant+hepatic+failure:+first+successful+case+report.">Auxiliary partial orthotopic liver&#8211;transplantation (APOLT) for fulminant hepatic failure: first successful case report.</a> World J Surg. 15, 660-666 (1991).</li><li>Erhard, J., Lange, R., Giebler, R., Rauen, U., de Groot, H., Eigler, F. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Arterialization+of+the+portal+vein+in+orthotopic+and+auxiliary+liver+transplantation.">Arterialization of the portal vein in orthotopic and auxiliary liver transplantation.</a> Transplantation. 60, 877-879 (1995).</li><li>Erhard, J., 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=Auxiliary+liver+transplantation+with+arterialization+of+the+portal+vein+for+acute+hepatic+failure.">Auxiliary liver transplantation with arterialization of the portal vein for acute hepatic failure.</a> Transpl Int. 11, 266-271 (1998).</li><li>Emond, J., Capron-Laudereau, M., Meriggi, F., Bernuau, J., Reynes, M., Houssin, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Extent+of+Hepatectomy+in+the+rat.">Extent of Hepatectomy in the rat.</a> Eur Surg Res. 21, 251-259 (1989).</li><li>Mas, A., Rode`s, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fulminant+hepatic+failure.">Fulminant hepatic failure.</a> Lancet. 349, 1081-1085 (1997).</li><li>Terpstra, O. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Auxiliary+liver+grafting:+a+new+concept+in+liver+transplantation.">Auxiliary liver grafting: a new concept in liver transplantation.</a> The Lancet. 342, 758 (1993).</li><li>Terpstra, O. T., Reuvers, C. B., Schalm, S. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Auxiliary+heterotopic+liver+transplantation.">Auxiliary heterotopic liver transplantation.</a> Transplantation. 45, 1003-1007 (1988).</li><li>Schleimer, K., Lange, R., Rauen, U., Erhard, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Auxiliary+liver+transplantation+in+acute+liver+failure+in+the+rat+-+an+illustrated+description+of+a+new+surgical+approach.">Auxiliary liver transplantation in acute liver failure in the rat - an illustrated description of a new surgical approach.</a> Langenbecks Arch Surg. 384 (2), 204-208 (1999).</li><li>Schleimer, 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=Physiologic+microcirculation+of+the+heterotopically+transplanted+rat+liver+with+portal+vein+arterialization+depending+on+optimal+stent+diameter.">Physiologic microcirculation of the heterotopically transplanted rat liver with portal vein arterialization depending on optimal stent diameter.</a> Med Sci Monit. 28 (BR140-145), (2006).</li><li>Hong, H. Q., 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=Study+of+portal+arterialization+with+auxiliary+liver+in+rats.">Study of portal arterialization with auxiliary liver in rats.</a> Hiroshima J. Med. Sci. 40, 29-33 (1991).</li><li>Higgins, G. M., Anderson, R. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Experimental+pathology+of+the+liver.+I--Restoration+of+the+liver+of+the+white+rat+following+partial+surgical+removal.">Experimental pathology of the liver. I--Restoration of the liver of the white rat following partial surgical removal.</a> Arch. Path. 12, 186-202 (1931).</li><li>Schleimer, 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=Auxiliary+liver+transplantation+with+flow-regulated+portal+vein+arterialization+offers+a+successful+therapeutic+option+in+acute+hepatic+failure--investigations+in+heterotopic+auxiliary+rat+liver+transplantation.">Auxiliary liver transplantation with flow-regulated portal vein arterialization offers a successful therapeutic option in acute hepatic failure--investigations in heterotopic auxiliary rat liver transplantation.</a> Transpl Int. 19 (7), 581-588 (2006).</li><li>Lee, S., Edgington, T. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Liver+transplantation+in+the+rat.">Liver transplantation in the rat.</a> Surg. Forum. 17, 220-222 (1966).</li><li>Hess, F., Willemen, A., Jerusalem, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Auxiliary+liver+transplantation+in+the+rat,+influence+of+the+condition+of+the+recipient's+liver+on+the+fate+of+the+graft.">Auxiliary liver transplantation in the rat, influence of the condition of the recipient's liver on the fate of the graft.</a> Eur. Surg. Res. 9, 270-279 (1977).</li><li>Aguirrezabalaga, J., Arnal, F., Marini, M., Centeno, A., Fernandez-Selles, C., Rey, I., Gomez, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Auxiliary+liver+transplantation+with+portal+arterialization+in+the+rat:+description+of+a+new+model.">Auxiliary liver transplantation with portal arterialization in the rat: description of a new model.</a> Microsurgery. 22, 21-26 (2002).</li><li>Palmes, D., Dietl, K. H., Drews, G., Holzen, J. P., Spiegel, H. U. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Auxiliary+partial+orthotopic+liver+transplantation:+treatment+of+acute+liver+failure+in+a+new+rat+model.">Auxiliary partial orthotopic liver transplantation: treatment of acute liver failure in a new rat model.</a> Langenbecks Arch Surg. (386), 534-541 (2002).</li><li>Schleimer, 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=Improved+microcirculation+of+a+liver+graft+by+controlled+portal+vein+arterialization.">Improved microcirculation of a liver graft by controlled portal vein arterialization.</a> J Surg Res. 116 (2), 202-210 (2004).</li><li>Schleimer, 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=Portal+hyperperfusion+causes+disturbance+of+microcirculation+and+increased+rate+of+hepatocellular+apoptosis:+investigations+in+heterotopic+rat+liver+transplantation+with+portal+vein+arterialization.">Portal hyperperfusion causes disturbance of microcirculation and increased rate of hepatocellular apoptosis: investigations in heterotopic rat liver transplantation with portal vein arterialization.</a> Transplant Proc. 38 (3), 725-729 (2006).</li></ol>

Todos os autores declararam interesses conflitantes.

O primeiro transplante de fígado de rato heterotópico auxiliar foi descrito por Lee et al. 16, em 1966, seguido por Hess. 17 A veia portal do enxerto anastomose com a veia portal do destinatário. Durante este processo, o fornecimento de sangue da veia porta para o fígado nativo é comprometida. Em 1991, Hong realizada parada com PVA através da artéria ilíaca do destinatário. 13 Sem regulação do fluxo sangüíneo portal enorme e congestionam...

Insuficiência hepática aguda representa um problema grave em medicina clínica que leva a uma taxa de mortalidade de 60-80%. 8 Na insuficiência hepática aguda, transplante hepático auxiliar é uma terapia alternativa promissora para superar a fase aguda. O fígado nativo é deixado para regenerar e segue para a regeneração dos medicamentos de imunossupressão pode ser interrompido. Como consequência, o enxerto se torne atrófica. No entanto, a técnica operatória para transplante de fígado auxiliar ainda é discutido. 1-3 Neste contexto, Terpstra relatou a falta de espaço para o enxerto, o sangue portal roubar fenômeno e que o problema de congestão venosa e drenagem 9-10 veia porta do doador. anastomose fim-a-lado a veia porta do destinatário. Isto resultou em competição de fluxo portal entre o enxerto eo fígado nativo e em trombose da veia porta. A maioria destes problemas técnicos podem ser evitadas com a utilização do método APOLT-(orthotopic implantação de segmentos auxiliares). 4 No entanto, este método requer extensas ressecção de ambos o fígado nativo e o enxerto. 4, em 1998, e a sua equipa Erhard desenvolvido o transplante heterotópico auxiliar fígado (PARADA), utilizando a arterialização da veia porta (PVA) (Figura 1 ). O hilo e a veia portal do fígado nativa sejam respeitados com a utilização do presente método. Isso poupa o destinatário de efeitos colaterais indesejáveis, tais como trombose da veia porta e sangue portal roubar fenômeno, que poderia perturbar a recuperação do fígado nativo. Além disso, há grandes ressecções hepáticas do fígado nativo e enxerto são necessários para este método, com conseqüente menor sangramento e mais massa total do parênquima hepático restante para colmatar (enxerto) e regeneração (fígado nativo). Esta técnica se mostrou promissora resultados clínicos iniciais. 5-6 O objetivo deste trabalho experimental foi o desenvolvimento de uma técnica de parar com Arterialização veia porta emo rato para fins de pesquisa (figura 2). Nos experimentos preliminares, não levamos em consideração a regulação do fluxo sangüíneo da veia portal arterializado, o que resultou em congestionamento do enxerto transplantado após a reperfusão e sangramento maciço da cápsula do fígado. 11 Desde então, o fluxo de sangue na veia portal arterializado foi regulada por um stent com um diâmetro interno de 0,3 mm e um comprimento de 8 mm. Experiências piloto tinham mostrado que um diâmetro de 0,3 milímetros de stent produz um fluxo de sangue média na veia porta arterializado que está dentro da gama fisiológica superior (1,7 +/- 0,4 ml / min / g de peso de fígado). Nossos 12 resultados sem regulação do fluxo sanguíneo correlacionar com os resultados de Hong, que realizou parada com PVA através da artéria ilíaca do destinatário. 13 Em seus experimentos, sem regulação do fluxo sangüíneo portal enorme e congestionamentos sinusoidais ocorreu. O objetivo do nosso vídeo é demonstrate passo a passo desta técnica cirúrgica experimental ne.w no rato. No longo prazo, a técnica proposta pode ser utilizado em estudos sobre a regeneração do fígado e da concorrência inter-fígado e Arterialização veia porta, hiperperfusï¿½ veia porta e regulação do fluxo sangüíneo portal.

<table border="0" cellpadding="0" cellspacing="0" width="573">
	<tbody>
		<tr height="41">
			<td height="41" style="height:41px;width:216px;">Braided Silk black 7-0&#160;</td>
			<td style="width:71px;">Ethicon Products</td>
			<td style="width:119px;">892100</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:216px;">Vicryl 3-0</td>
			<td style="width:71px;">Ethicon Products</td>
			<td style="width:119px;">J285G</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:216px;">Vicryl 4-0</td>
			<td style="width:71px;">Ethicon Products</td>
			<td style="width:119px;">J284G</td>
			<td style="width:167px;"></td>
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		<tr height="41">
			<td height="41" style="height:41px;width:216px;">Ethilon 8-0</td>
			<td style="width:71px;">Ethicon Products</td>
			<td style="width:119px;">1714G</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:216px;">Ethilon 10-0</td>
			<td style="width:71px;">Ethicon Products</td>
			<td style="width:119px;">7770G</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Vasofix safety 14G</td>
			<td style="width:71px;">B Braun</td>
			<td style="width:119px;">2758853</td>
			<td style="width:167px;"></td>
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		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Vasofix safety 20G</td>
			<td style="width:71px;">B Braun</td>
			<td style="width:119px;">2758818</td>
			<td style="width:167px;"></td>
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		<tr height="201">
			<td height="201" style="height:201px;width:216px;">2 French (.012&#8221;/0.3mm ID x .025&#8221;/0.6mm OD) x 24&#8221;/60cm polyurethane catheter</td>
			<td style="width:71px;">Access technologies a division of Norfolk Medical, Inc., Illinois, USA</td>
			<td style="width:119px;">CNC-2P</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="119">
			<td height="119" style="height:119px;width:216px;">HTK solution,</td>
			<td rowspan="2" style="width:71px;">Dr. Franz K&#246;hler Chemie GmbH, Alsbach-H&#228;hnlein, Germany</td>
			<td rowspan="2" style="width:119px;"></td>
			<td rowspan="2" style="width:167px;"></td>
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			<td height="21" style="height:21px;width:216px;">Custodiol</td>
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			<td height="22" style="height:22px;width:216px;">Isofluorane 5 %</td>
			<td style="width:71px;"></td>
			<td style="width:119px;"></td>
			<td style="width:167px;">Any genericon</td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:216px;">Heparin</td>
			<td style="width:71px;"></td>
			<td style="width:119px;"></td>
			<td style="width:167px;">Any genericon</td>
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			<td height="22" style="height:22px;width:216px;">0.9% saline</td>
			<td style="width:71px;"></td>
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			<td style="width:167px;">Any genericon</td>
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			<td height="22" style="height:22px;width:216px;">Buprenorphine</td>
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heterotopic auxiliary rat liver transplantation with flow-regulated portal vein arterialization in acute hepatic failure

Na insuficiência hepática de transplante de fígado auxiliar aguda é uma abordagem alternativa interessante. O objetivo é fornecer um apoio temporário até que o fígado nativo não regenerou. 1-3 O APOLT-método, a implantação ortotópico de segments- auxiliar evita a maioria dos problemas técnicos. No entanto, este método necessita de grandes ressecções de ambos o fígado nativo e o enxerto. 4 Em 1998, Erhard desenvolvido o transplante heterotópico auxiliar fígado (PARADA), utilizando a arterialização da veia porta (PVA) (Figura 1). Esta técnica se mostrou promissora resultados clínicos iniciais. 5-6 Desenvolvemos uma parada-técnica com fluxo regulado PVA no rato para examinar a influência do PVA-fluxo regulado na morfologia e função (Figura 2) enxerto. Um enxerto de fígado reduzida para 30% do seu tamanho original, foi heterotopicamente implantado na região renal direita do receptor após o explante do rim direito.60; A veia cava de infra-hepática do enxerto anastomose com a veia cava infrahepatic do destinatário. O Arterialização da veia porta do doador foi realizado via artéria renal direita do destinatário com a técnica de stent. A regulação do fluxo de sangue da veia porta arterializado foi conseguido com a utilização de uma prótese com um diâmetro interno de 0,3 mm. O tronco celíaco do enxerto foi de ponta a lado com anastomose aorta do destinatário e do ducto biliar foi implantado no duodeno. A ressecção do fígado nativo foi realizada para induzir falha hepática aguda 7. Deste modo foram realizados 112 transplantes. A taxa de sobrevivência perioperatória foi de 90% e a taxa de sobrevivência de 6 semanas foi de 80%. Seis semanas após a operação, o fígado nativo regenerada, que apresenta um aumento de peso de 2,3 ± 0,8 g para 9,8 ± 1 g. Nesta altura, o peso do enxerto diminuiu de 3,3 ± 0,8 g para 2,3 ± 0,8 g. </p&gt; Nós fomos capazes de obter promissores resultados a longo prazo em termos de morfologia e função do enxerto. HALT com fluxo regulado PVA preenche de forma confiável insuficiência hepática aguda até o fígado nativo regenera.

A duração média do transplante foi de cerca de 150 min. O tempo de isquemia fria foi de 64 +/- 7 min, o tempo de isquemia quente foi inferior a 25 min. A taxa de sobrevivência perioperatória foi de 90% e a taxa de sobrevivência de 6 semanas foi de 80%. A perda de 20% foi devido a peritonite por causa do deslocamento do stent e extravasamento biliar (quatro animais) durante a primeira semana de pós-operatório e, devido à pneumonia, abcesso intra-abdominal e íleo (sete animais) duran...

Watch this Scientific Journal Video about Heterotopic Auxiliary Rat Liver Transplantation With Flow-regulated Portal Vein Arterialization in Acute Hepatic Failure at JoVE.com

Heterotopic Auxiliary Rat Liver Transplantation With Flow-regulated Portal Vein Arterialization in Acute Hepatic Failure

O transplante de fígado auxiliar proporciona um suporte temporário em insuficiência hepática aguda, até que a regeneração do fígado, não. O transplante heterotópico auxiliar fígado (HALT) com Arterialização veia porta (PVA) torna a função hepática está suficientemente bem. Desenvolvemos uma técnica análoga, no rato, para examinar a influência da arterialização veia portal na morfologia e função do enxerto.

Heterotópico Rat auxiliar Liver Transplantation com fluxo regulado Portal Vein Arterialização na Insuficiência Hepática Aguda

In acute hepatic failure auxiliary liver transplantation is an interesting alternative approach. The aim is to provide a temporary support until the ...

heterotopic-auxiliary-rat-liver-transplantation-with-flow-regulated

Research

JoVE Journal

Medicine

12.6K visualizações.  University Hospital RWTH Aachen.  O transplante de fígado auxiliar proporciona um suporte temporário em insuficiência hepática aguda, até que a regeneração do fígado, não. O transplante heterotópico auxiliar fígado (HALT) com Arterialização veia porta (PVA) torna a função hepática está suficientemente bem. Desenvolvemos uma técnica análoga, no rato, para examinar a influência da arterialização veia portal na morfologia e função do enxerto. 

Vídeo: Heterotopic Auxiliary Rat Liver Transplantation With Flow-regulated Portal Vein Arterialization in Acute Hepatic Failure

Surgical Procedures for a Rat Model of Partial Orthotopic Liver Transplantation with Hepatic Arterial Reconstruction

Orthotopic liver transplantation (OLT) in rats using a whole or partial graft is an indispensable experimental model for transplantation research, such as studies on graft preservation and ischemia-reperfusion injury 1,2, immunological responses 3,4, hemodynamics 5,6, and small-for-size syndrome 7. The rat OLT is among the most difficult animal models in experimental surgery and demands advanced microsurgical skills that take a long time to learn. Consequently, the use of this model has been limited. Since the reliability and reproducibility of results are key components of the experiments in which such complex animal models are used, it is essential for surgeons who are involved in rat OLT to be trained in well-standardized and sophisticated procedures for this model.
While various techniques and modifications of OLT in rats have been reported 8 since the first model was described by Lee et al. 9 in 1973, the elimination of the hepatic arterial reconstruction 10 and the introduction of the cuff anastomosis technique by Kamada et al. 11 were a major advancement in this model, because they simplified the reconstruction procedures to a great degree. In the model by Kamada et al., the hepatic rearterialization was also eliminated. Since rats could survive without hepatic arterial flow after liver transplantation, there was considerable controversy over the value of hepatic arterialization. However, the physiological superiority of the arterialized model has been increasingly acknowledged, especially in terms of preserving the bile duct system 8,12 and the liver integrity 8,13,14.
In this article, we present detailed surgical procedures for a rat model of OLT with hepatic arterial reconstruction using a 50% partial graft after ex vivo liver resection. The reconstruction procedures for each vessel and the bile duct are performed by the following methods: a 7-0 polypropylene continuous suture for the supra- and infrahepatic vena cava; a cuff technique for the portal vein; and a stent technique for the hepatic artery and the bile duct.

Orthotopic liver transplantation in rats is an indispensable experimental model for biomedical research. Here we present our surgical procedures for orthotopic rat liver transplantation with hepatic arterial reconstruction using a 50% partial graft.

Procedimentos cirúrgicos para um modelo de rato de Transplante de Fígado parcial ortotópico com reconstrução arterial hepática

Orthotopic liver transplantation (OLT) in rats using a whole or partial graft is an indispensable experimental model for transplantation research, such as ...

Medicina

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.

Rat heterotópico abdominal coração / pulmão Transplante Single-carregado em uma configuração-Volume

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

Heterotopic Cervical Heart Transplantation in Mice

The heterotopic cervical heart transplantation in mice is a valuable tool in transplant and cardiovascular research. The cuff technique greatly simplifies this model by avoiding challenging suture anastomoses of small vessels thereby reducing warm ischemia time. In comparison to abdominal graft implantation the cervical model is less invasive and the implanted graft is easily accessible for further follow-up examinations. Anastomoses are performed by pulling the ascending aorta of the graft over the cuff with the recipient&#8217;s common carotid artery and by pulling the main pulmonary artery over the cuff with the external jugular vein. Selection of appropriate cuff size and complete mobilization of the vessels are important for successful revascularization. Ischemia-reperfusion (I/R) injury can be minimized by perfusing the graft with a cardioplegic solution and by hypothermia. In this article, we provide technical details for a simplified and improved cuff technique, which should allow surgeons with basic microsurgical skills to perform the procedure with a high success rate.

The cuff technique shortens and facilitates the model of heterotopic cervical heart transplantation in mice by avoiding technically challenging suture anastomoses of small vessels. The technical details shown in this video paper should allow researches to establish this model in their laboratories.

Heterotopic Cervical Transplante Cardíaco em Ratos

The heterotopic cervical heart transplantation in mice is a valuable tool in transplant and cardiovascular research. The cuff technique greatly simplifies ...

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.

Modelo do rato da partição fígado associando e ligadura da veia porta para procedimento encenado Hepatectomy (ALPPS)

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

Re-Arterialized Rat Partial Liver Transplantation with an in vivo Vessel-Oriented 70% Hepatectomy

Split liver transplantation and living liver donor liver transplantation were developed in the clinic to utilize liver organs in a more efficient manner. To better understand the mechanism behind these surgical procedures, a rat partial liver transplantation (PLTx) model was established for relevant surgical studies. Because of the complexity of the rat PLTx model, a protocol with detailed descriptions is required. An article published previously reported a protocol in which ex vivo hepatectomy was used to achieve 50% rat PLTx. In contrast to this protocol, we introduced a re-arterialized PLTx with an in vivo 70% hepatectomy. An updated vessel-oriented hepatectomy was incorporated into the rat PLTx to refine the microsurgical procedure. The portal veins and hepatic arteries of the left lateral lobe and the median lobe were individually dissected and ligated before removal of the liver parenchyma, thereby decreasing the probability of bleeding in the remnant liver stump. Furthermore, an end-to-side vessel anastomosis between the common hepatic artery and the enlarged proper hepatic artery was introduced to re-arterialize the hepatic artery. By using this end-to-side vessel anastomosis technique, the diameter of the anastomosis was enlarged, thereby decreasing the difficulty of hand suture and maintaining a high rate of anastomotic patency. Moreover, the cuff anastomosis of the infrahepatic vena cava was slightly modified. A section of circumferential liver parenchyma around the vena cava of a recipient was preserved during cuff anastomosis to maintain the three-dimensional shape of the vascular lumen. This section of liver parenchyma was removed after completing the anastomosis. With this modification, the step involving placement of stay sutures was omitted, thereby further shortening the cuff anastomosis time. By using this protocol of rat PLTx, a low liver enzyme level, an intact liver lobular architecture and a high survival rate were achieved after microsurgery.

Re-Arterialized Rat Partial Liver Transplantation with an in vivo Vessel-Oriented 70% Hepatectomy

Here, a protocol involving re-arterialized rat partial liver transplantation is presented. Specifically, 70% liver was resected in vivo by using an updated technique of vessel-oriented hepatectomy. The hepatic artery was reconstructed in an end-to-side manner. The cuff technique was modified to shorten the anastomosis time of the infrahepatic vena cava.

Re-Arterialized transplante de fígado parcial de rato com na vivo orientado a embarcação 70% Hepatectomy

Split liver transplantation and living liver donor liver transplantation were developed in the clinic to utilize liver organs in a more efficient manner. ...

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.

Caracterização hemodinâmica invasiva da síndrome hipertensiva-Portal em ratos cirrose

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

Generation of a Rat Model of Acute Liver Failure by Combining 70% Partial Hepatectomy and Acetaminophen

Acute liver failure (ALF) is a clinical condition caused by various etiologies resulting in the loss of metabolic, biochemical, synthesizing, and detoxifying functions of the liver. In most irreversible liver damage cases, orthotropic liver transplant (OLT) remains the only available treatment. To study the therapeutic potential of a treatment for ALF, its prior testing in an animal model of ALF is essential. In the current study, an ALF model in rats was developed by combining 70% partial hepatectomy (PHx) and injections of acetaminophen (APAP) that provides a therapeutic window of 48 h. The median and left lateral lobes of the liver were removed to excise 70% of the liver mass and APAP was given 24 h postsurgically for 2 days. Survival in ALF-induced animals was found to be severely decreased. The development of ALF was confirmed by altered serum levels of the enzymes alanine amino transferase (ALT), aspartate amino transferase (AST), alkaline phosphatase (ALP); changes in prothrombin time (PT); and assessment of the international normalized ratio (INR). Study of the gene expression profile by qPCR revealed an increase in expression levels of genes involved in apoptosis, inflammation, and in the progression of liver injury. Diffused degeneration of hepatocytes and infiltration of immune cells was observed by histological evaluation. The reversibility of ALF was confirmed by the restoration of survival and serum levels of ALT, AST, and ALP after intrasplenic transplantation of syngeneic healthy rat hepatocytes. This model presents a reliable alternative to the available ALF animal models to study the pathophysiology of ALF as well as to evaluate the potential of a novel therapy for ALF. The use of two different approaches also makes it possible to study the combined effect of physical and drug-induced liver injury. The reproducibility and feasibility of current procedure is an added benefit of the model.

The acute liver failure animal model developed in the current study presents a feasible alternative for the study of potential therapies. The current model employs the combined effect of physical and drug-induced hepatic injury and provides a suitable time window to study the potential of novel therapies.

Geração de um modelo de rato de insuficiência hepática aguda, combinando 70% hepatectomia parcial e paracetamol

Acute liver failure (ALF) is a clinical condition caused by various etiologies resulting in the loss of metabolic, biochemical, synthesizing, and ...

Minimizing Post-Infusion Portal Vein Bleeding during Intrahepatic Islet Transplantation in Mice

Although the liver is currently accepted as the primary transplantation site for human islets in clinical settings, islets are transplanted under the kidney capsule in most rodent preclinical islet transplantation studies. This model is commonly used because murine intrahepatic islet transplantation is technically challenging, and a high percentage of mice could die from surgical complications, especially bleeding from the injection site post-transplantation. In this study, two procedures that can minimize the incidence of post-infusion portal vein bleeding are demonstrated. The first method applies an absorbable hemostatic gelatin sponge to the injection site, and the second method involves penetrating the islet injection needle through the fat tissue first and then into the portal vein by using the fat tissue as a physical barrier to stop bleeding. Both methods could effectively prevent bleeding-induced mouse death. The whole liver section showing islet distribution and evidence of islet thrombosis post-transplantation, a typical feature for intrahepatic islet transplantation, were presented. These improved protocols refine the intrahepatic islet transplantation procedures and may help laboratories set up the procedure to study islet survival and function in pre-clinical settings.

Here we present refined surgical procedures on successfully performing intraportal islet transplantation, a clinically relevant but technically challenging surgical procedure, in mice.

Minimizando o sangramento da veia do portal pós-infusão durante o transplante de ilhotas intrahápticas em camundongos

Although the liver is currently accepted as the primary transplantation site for human islets in clinical settings, islets are transplanted under the ...

Rat Model of Normothermic Ex-Situ Perfused Heterotopic Heart Transplantation

Heart transplantation is the most effective therapy for end-stage heart failure. Despite the improvements in therapeutic approaches and interventions, the number of heart failure patients waiting for transplantation is still increasing. The normothermic ex situ preservation technique has been established as a comparable method to the conventional static cold storage technique. The main advantage of this technique is that donor hearts can be preserved for up to 12 h in a physiologic condition. Moreover, this technique allows resuscitation of the donor hearts after circulatory death and applies required pharmacologic interventions to improve donor function after implantation. Numerous animal models have been established to improve normothermic ex situ preservation techniques and eliminate preservation-related complications. Although large animal models are easy to handle compared to small animal models, it is costly and challenging. We present a rat model of normothermic ex situ donor heart preservation followed by heterotopic abdominal transplantation. This model is relatively cheap and can be accomplished by a single experimenter.

Here, we present an assessment protocol of a heterotopically implanted heart after normothermic ex situ preservation in the rat model.

Modelo de Transplante Cardíaco Heterotópico Perfundido Normothermic Ex-Situ em Rato

Heart transplantation is the most effective therapy for end-stage heart failure. Despite the improvements in therapeutic approaches and interventions, the ...

Transplante de coração in vivo para estudar a transição endotelial para mesenquimal

A Neonatal Heterotopic Rat Heart Transplantation Model for the Study of Endothelial-to-Mesenchymal Transition

Endocardial fibroelastosis (EFE), defined by subendocardial tissue accumulation, has major impacts on the development of the left ventricle (LV) and precludes patients with congenital critical aortic stenosis and hypoplastic left heart syndrome (HLHS) from curative anatomical biventricular surgical repair. Surgical resection is currently the only available therapeutic option, but EFE often recurs, sometimes with an even more infiltrative growth pattern into the adjacent myocardium.
To better understand the underlying mechanisms of EFE and to explore therapeutic strategies, an animal model suitable for preclinical testing was developed. The animal model takes into consideration that EFE is a disease of the immature heart and is associated with flow disturbances, as supported by clinical observations. Thus, the heterotopic heart transplantation of neonatal rat donor hearts is the basis for this model.
A neonatal rat heart is transplanted into an adolescent rat&#39;s abdomen and connected to the recipient&#39;s infrarenal aorta and inferior vena cava. While perfusion of the coronary arteries preserves the viability of the donor heart, flow stagnation within the LV induces EFE growth in the very immature heart. The underlying mechanism of EFE formation is the transition of endocardial endothelial cells to mesenchymal cells (EndMT), which is a well-described mechanism of early embryonic development of the valves and septa but also the leading cause of fibrosis in heart failure. EFE formation can be macroscopically observed within days after transplantation. Transabdominal echocardiography is used to monitor the graft viability, contractility, and the patency of the anastomoses. Following euthanasia, the EFE tissue is harvested, and it shows the same histopathological characteristics as human EFE tissue from HLHS patients.
This in vivo model allows for studying the mechanisms of EFE development in the heart and testing treatment options to prevent this pathological tissue formation and provides the opportunity for a more generalized examination of EndMT-induced fibrosis.

Autor em destaque: Um modelo de transplante cardíaco de rato heterotópico neonatal para o estudo da transição endotelial para mesenquimal  

This work presents an animal model of endothelial-to-mesenchymal transition-induced fibrosis, as seen in congenital cardiac defects such as critical aortic stenosis or hypoplastic left heart syndrome, which allows for detailed histological tissue evaluation, the identification of regulatory signaling pathways, and the testing of treatment options.

Modelo de Transplante Cardíaco Heterotópico Neonatal em Rato para Estudo da Transição Endotelial-Mesenquimal

Endocardial fibroelastosis (EFE), defined by subendocardial tissue accumulation, has major impacts on the development of the left ventricle (LV) and ...