Questo studio è stato finanziato dalla &quot;Deutsche Forschungsgemeinschaft&quot; (DFG). K. Schleimer è stato sostenuto dalla Lise Meitner Fellowship del Ministero della Scienza e della Ricerca del NRW. Vorremmo ringraziare la signora Maria-Giovanna Blümich per l&#39;eccellente assistenza editing. Dr. Maria Kokozidou è stato finanziato da Fresenius 2011_A61.

1. Gli animali sperimentali <ol><li> Ottenere l&#39;approvazione Comitato Etico Locale per tutti gli esperimenti che verranno eseguite, in conformità con le linee guida di protezione degli animali FELASA (K 40, 17/99). </li><li> Utilizzare i ratti Lewis maschi disponibili per l&#39;acquisto da qualsiasi allevatore commerciale (ad esempio, Charles River Wiga GmbH, Sulzfeld, Germania). Qui, 224 animali (112 donatori, 112 destinatari) sono stati utilizzati per eseguire 112 trapianti. I destinatari sono stati sacrificati a 6 settimane dopo l&#39;intervento. Al momento dell&#39;operazione, gli animali dovrebbero pesare 250-350 g. </li><li> Tenere gli animali in ambienti climatizzati con libero accesso a cibo e acqua. </li></ol> 2 Anestesia e preparazione operativa <ol><li> Posizionare il ratto in una scatola di plexiglas collegato alla macchina per anestesia con un tasso di flusso di ossigeno di 1 L / min, e un vaporizzatore isofluorane con il 5% di gas anestetico per indurre l&#39;anestesia. Mantenere l&#39;anestesia al 1,5% isofluorane. </li><li> Mettere i topi inposizione supina su una piastra riscaldante. </li><li> Rimuovere il pelo del ventre con una crema depilazione. </li><li> Preparare un campo chirurgico sterile come segue: Utilizzare un disinfettante per pulire il banco. Per la disinfezione del sito di incisione uso Clorexidina prima, seconda alcool e, infine, una soluzione scrub (ad esempio, clorexidina e il 70% di alcool isopropilico). Indossare un camice chirurgico monouso e guanti chirurgici. </li><li> Utilizzare strumenti di microchirurgia sterili. </li><li> Utilizzare un microscopio funzionamento in 12-20 volte di ingrandimento. </li></ol> 3 Procedura chirurgica <ol><li> Funzionamento dei donatori Preparazione <ol><li> Eseguire una laparotomia mediana e resecare il lobo laterale sinistro e il lobo mediano del trapianto, come descritto da Higgins. 14 </li><li> Poi preparare il legamento epatoduodenale. Prima legare il dotto biliare per indurre congestione e dilatazione del piccolo dotto biliare. Poi legare e transetto la gastroduodenale vena e arteria. Successivamente, isolare il tronco celiaco e l&#39;arteria epatica comune e transetto l&#39;arteria gastrica sinistra e l&#39;arteria splenica. Incidere il dotto biliare congestionato, e di introdurre uno stent 20-G in esso. Il posizionamento dello stent deve garantire il drenaggio dell&#39;intero sistema biliare ilare. Transetto il dotto biliare distalmente allo stent. </li><li> Isolare il segmento infraepatica della vena cava proprio sopra la vena renale destra. Perfusione </li><li> Dopo legatura dell&#39;aorta sopra il tronco celiaco, bloccare la vena porta e incidere esso. Introdurre il catetere perfusione (14 G) nella vena porta, ed eseguire la perfusione con 30 ml di istidina-triptofano-chetoglutarato (HTK) -solution (12 cm H2O, 4 ° C). </li><li> Tagliare il diaframma e la vena cava sovraepatica per consentire al perfusato di fuoriuscire. </li><li> Accise il tronco celiaco con una patch aortica e sciacquare con soluzione istidina-triptofano-chetoglutarato. Espianto </li><li> Transetto la vena cava infraepatica proprio sopra la vena renale destra. </li><li> Quindi, collocare uno stent con un diametro di 0,3 mm nella vena porta e fissarlo con una legatura. Durante la seguente operazione, la vena porta viene arterializzato attraverso l&#39;arteria renale destra. Transetto la vena porta distalmente allo stent. </li><li> Dopo la legatura della vena cava sovraepatica, transetto insieme con un polsino di diaframma. </li><li> Espiantare l&#39;innesto e conservarla in istidina-triptofano-chetoglutarato (HTK) -solution a 4 ° C in un sacchetto di plastica sul ghiaccio. Suture Luogo d&#39;angolo (8-0 Ethilon) nella vena cava infraepatica. </li></ol></li><li> Funzionamento del destinatario <ol><li> Eseguire una laparotomia mediana e preparare la vena cava infraepatica. Quindi isolare l&#39;arteria renale destra e l&#39;uretere, legare l&#39;uretere e transetto esso. Bloccare l&#39;arteria renale destra alla sua origine, quindi legare la vena renale destra. Poi, dopo la recisione di entrambi rivasi nali, eseguire la nefrectomia. Successivamente, ridurre l&#39;arteria renale destra, sciacquare con soluzione salina eparina (soluzione salina di eparina 40 UI / ml). </li><li> Transetto della vena renale destra prossimale alla legatura dopo il bloccaggio prossimale e distale della vena cava infraepatica per creare un ovale lume della vena cava infraepatica. Sciacquare la vena cava con una soluzione salina eparina. Impianto del trapianto nella regione renale destra dell&#39;addome del destinatario. </li><li> Anastomose end-to-side vena cava infraepatica del donatore al recipient`s vena cava con un 8-0 Ethilon sutura continua. </li><li> Dopo il risciacquo con soluzione di eparina / salina, introdurre lo stent nella vena porta in arteria renale destra (Figura 3). Collegare la vena porta e l&#39;arteria renale con 2 punti (8-0 Ethilon). Fissare lo stent collegati dell&#39;arteria renale con una legatura (intrecciato di seta nera 7-0). </li><li> Per consentire la riperfusione, prima rimuovere la pinza arteriosa, allora entrambimorsetti venosi (Figura 4). </li><li> Preparare l&#39;aorta sottorenale, quindi inserire un prossimale e distale microclamp e incidere l&#39;aorta destra tra i due morsetti. Eseguire una anastomosi termino-laterale tra l&#39;aorta e il tronco celiaco del donatore, utilizzando una patch aortica, con un 10-0 Ethilon sutura continua. </li><li> Inserire il dotto biliare nel duodeno: Applicare una sutura a borsa nella parete duodenale, fare una piccola incisione nella parete duodenale proprio nel mezzo della a borsa. Introdurre il dotto biliare stent nel duodeno, e stringere la sutura a borsa. Infine, tenere premuto il duodeno e il dotto biliare strettamente insieme con due punti (Ethilon 8-0) (Figura 5). </li><li> Quindi eseguire una resezione subtotale del fegato nativo, prima descritto da Emond et al. per imitare insufficienza epatica acuta. 7 </li><li> Fissare la vena cava sovraepatica dell&#39;innesto, con un polsino di diaframma, alla parete addominale laterale del recipient con 4-0 Vicryl sutura. Risciacquare l&#39;addome con soluzione salina (Figura 6). </li><li> Per lo strato muscolare utilizzare una sutura continua 4-0 Vicryl e per la sutura cutanea una esecuzione 3-0 Vicryl. </li></ol></li></ol> 4 Recupero e gestione postoperatoria <ol><li> Lasciate che gli animali recuperare da anestesia sotto una lampada di riscaldamento a raggi infrarossi. </li><li> Iniettare Buprenorfina alla dose di 0,03-0,05 mg / kg di peso corporeo sc ogni 8-12 ore per 2 giorni. </li><li> Esaminare gli animali concernenti la condizione generale e segni di malattia o di infezioni. </li></ol> 5 Calcolo del peso iniziale Native Fegato Calcolare il peso iniziale fegato nativo nel seguente modo: sottrarre il peso dei lobi epatici asportati dal peso del fegato completa (2,66% del peso corporeo (questo valore con una deviazione standard di +/- 0,49% è stata determinata in esperimenti precedenti con n = 80 animali dello stesso ceppo ege)). 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>

Tutti gli autori hanno dichiarato senza interessi in competizione.

Il primo trapianto di fegato di ratto ausiliario eterotopico è stato descritto da Lee et al., 16 nel 1966, seguita da Hess. 17 La vena porta del graft è stato anastomizzata con la vena porta del destinatario. Durante questa procedura, il portale afflusso di sangue venoso al fegato nativo è compromessa. Nel 1991 Hong eseguito HALT con PVA attraverso l&#39;arteria iliaca del destinatario. 13 senza regolazione del flusso sanguigno massiccio portale e...

Insufficienza epatica acuta pone un problema grave in medicina clinica che porta a un tasso di mortalità del 60-80%. 8 In caso di insufficienza epatica acuta, trapianto di fegato ausiliario è una terapia alternativa promettente per colmare la fase acuta. Il fegato nativo è lasciato per rigenerare e segue alla rigenerazione dei farmaci immunosoppressione possono essere sospesi. Come conseguenza l&#39;innesto diventerà atrofica. Tuttavia, la tecnica operatoria per il trapianto di fegato ausiliario è ancora discussa. 1-3 In questo contesto, Terpstra ha segnalato la mancanza di spazio per il trapianto, il sangue portale rubare fenomeno e il problema della congestione venosa e il drenaggio. 9-10 vena porta del donatore è stato anastomizzato end-to-side alla vena porta del destinatario. Ciò ha provocato una concorrenza portale flusso tra l&#39;innesto e il fegato nativo e trombosi della vena porta. La maggior parte di questi problemi tecnici possono essere evitati con l&#39;uso del metodo APOLT-(orthotopic impianto di segmenti ausiliari). 4 Tuttavia, questo metodo richiede ampie resezioni sia del fegato nativo e l&#39;innesto. 4 Nel 1998, Erhard e il suo team hanno sviluppato il trapianto eterotopico ausiliario di fegato (HALT) utilizzando arterialization vena porta (PVA) (Figura 1 ). L&#39;ilo e la vena porta del fegato nativo sono rispettati con l&#39;uso di questo metodo. Che non risparmia il destinatario da effetti collaterali indesiderati come la trombosi della vena porta e sangue portale rubare fenomeno, che potrebbe disturbare il recupero del fegato nativo. Inoltre, non estese resezioni epatiche del fegato nativo e l&#39;innesto sono necessari per questo metodo, con conseguente minor sanguinamento e più restante massa totale del parenchima epatico per colmare (trapianto) e rigenerazione (fegato nativo). Questa tecnica ha mostrato promettenti risultati clinici iniziali. 5-6 L&#39;obiettivo di questo lavoro sperimentale è stato lo sviluppo di una tecnica di HALT con arterialization vena porta inratto per la ricerca (Figura 2). Negli esperimenti preliminari non abbiamo preso in considerazione la regolazione del flusso sanguigno del portale arterializzato vena, che ha provocato la congestione del graft trapiantato dopo riperfusione e sanguinamento massiccio dalla capsula epatica. 11 Da allora il flusso di sangue nella vena porta arterializzato è regolata da uno stent con un diametro interno di 0,3 mm e una lunghezza di 8 mm. Esperimenti pilota hanno dimostrato che un diametro stent di 0,3 mm produce un flusso medio di sangue nella vena porta arterializzato che è all&#39;interno del range fisiologico superiore (1,7 +/- 0,4 ml / min / g di peso del fegato). 12 I nostri risultati senza regolazione del flusso sanguigno correlare con i risultati di Hong, che ha eseguito HALT con PVA attraverso l&#39;arteria iliaca del destinatario. 13 Nei suoi esperimenti senza regolazione del flusso sanguigno massiccio portale e si è verificato congestioni sinusoide. Lo scopo del nostro video è quello demonstrate passo passo questa Ne.w tecnica chirurgica sperimentale nel ratto. A lungo termine la tecnica proposta può essere utilizzata negli studi sulla rigenerazione del fegato e la concorrenza tra fegato e arterialization vena porta, iperperfusione vena porta e la regolazione del flusso sanguigno portale.

<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>
		</tr>
		<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>
		</tr>
		<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>
		</tr>
		<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>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Custodiol</td>
		</tr>
		<tr height="22">
			<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>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:216px;">0.9% saline</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;">Buprenorphine</td>
			<td style="width:71px;"></td>
			<td style="width:119px;"></td>
			<td style="width:167px;">Any genericon</td>
		</tr>
	</tbody>
</table>

heterotopic auxiliary rat liver transplantation with flow-regulated portal vein arterialization in acute hepatic failure

Nel trapianto di fegato ausiliario insufficienza epatica acuta è un approccio alternativo interessante. L&#39;obiettivo è quello di fornire un supporto temporaneo fino alla mancanza fegato nativo ha rigenerato. 1-3 Il APOLT-metodo, l&#39;impianto ortotopico di segments- ausiliario evita la maggior parte dei problemi tecnici. Tuttavia, questo metodo richiede ampie resezioni sia del fegato nativo e l&#39;innesto. 4 Nel 1998, Erhard ha sviluppato il trapianto eterotopico ausiliario di fegato (HALT) utilizzando arterialization vena porta (PVA) (Figura 1). Questa tecnica ha mostrato promettenti risultati clinici iniziali. 5-6 Abbiamo sviluppato un HALT-tecnica con PVA-flusso regolato nel ratto per esaminare l&#39;influenza di PVA-flusso regolato sulla morfologia e funzione (Figura 2) trapianto. Un trapianto di fegato ridotto al 30% della sua dimensione originale, è stato heterotopically impiantato nella regione renale destra del destinatario dopo espianto del rene destro.60; La vena cava infra-epatica dell&#39;innesto è stata anastomizzata con la vena cava infraepatica del destinatario. La arterialization della vena porta del donatore è stata effettuata attraverso l&#39;arteria renale destra del destinatario con la tecnica dello stent. La regolazione del flusso sanguigno della vena porta arterializzato è stato ottenuto con l&#39;uso di uno stent con un diametro interno di 0,3 mm. Il tronco celiaco dell&#39;innesto era end-to-side anastomizzato con aorta del destinatario e il dotto biliare è stato impiantato nel duodeno. Una resezione subtotale del fegato nativo è stata eseguita per indurre insufficienza epatica acuta. 7 In questo modo sono stati eseguiti 112 trapianti. Il tasso di sopravvivenza perioperatoria è stata del 90% e il tasso di sopravvivenza di 6 settimane è stata dell&#39;80%. Sei settimane dopo l&#39;intervento, il fegato nativo rigenerato, mostrando un aumento di peso da 2,3 ± 0,8 g di 9,8 ± 1 g. In questo momento, il peso del graft è diminuito da 3,3 ± 0,8 g di 2,3 ± 0,8 g. </p&gt; Siamo stati in grado di ottenere risultati promettenti a lungo termine in termini di morfologia e funzione del trapianto. HALT-PVA con flusso regolato colma affidabile insufficienza epatica acuta fino a quando il fegato nativo rigenera.

La durata media del trapianto era di circa 150 min. Il tempo di ischemia fredda era 64 +/- 7 minuti, il tempo di ischemia calda era a meno di 25 min. Il tasso di sopravvivenza perioperatoria è stata del 90% e il tasso di sopravvivenza di 6 settimane è stata dell&#39;80%. La perdita del 20% è dovuto a peritonite a causa della dislocazione dello stent e la perdita biliare (quattro animali) durante la prima settimana postoperatoria e causa di polmonite, ascesso intra-addominale e ileo (sette...

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

Trapianto di fegato ausiliario fornisce un supporto temporaneo in insufficienza epatica acuta, fino a quando la rigenerazione del fegato fallire. Il trapianto eterotopico ausiliario di fegato (HALT) con arterialization vena porta (PVA) rende del fegato è sufficiente. Abbiamo sviluppato una tecnica analoga nel ratto, di esaminare l&#39;influenza del arterialization vena porta sulla morfologia e la funzione del trapianto.

Eterotopica ausiliario Ratto trapianto di fegato con flusso regolato Portal Vein arterialization in acuta insufficienza epatica

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 Views.  University Hospital RWTH Aachen.  Trapianto di fegato ausiliario fornisce un supporto temporaneo in insufficienza epatica acuta, fino a quando la rigenerazione del fegato fallire. Il trapianto eterotopico ausiliario di fegato (HALT) con arterialization vena porta (PVA) rende del fegato è sufficiente. Abbiamo sviluppato una tecnica analoga nel ratto, di esaminare l'influenza del arterialization vena porta sulla morfologia e la funzione del trapianto. 

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

Orthotopic Liver Transplantation in Rats

Clinical progress in the field of liver transplantation has been largely supported by animal models1,2. Since the publication of the first orthotopic rat liver transplantation in 1979 by Kamada et al.3, this model has remained the gold standard despite various proposed alternative techniques4. Nevertheless, its broader use is limited by its steep learning curve5.
In this video paper, we show a simple and easy-to-establish revision of Kamada's two-cuff technique. The suprahepatic vena cava anastomosis is performed manually with a running suture, and the vena porta and infrahepatic vena cava anastomoses are performed utilizing a quick-linker cuff system6. Manufacturing the quick-linker kit is shown in a separate video paper.

We present an easy-to-establish revision of the classical two-cuff technique for orthotopic liver transplantation in rat.

Trapianto di fegato in ratti

Clinical progress in the field of liver transplantation has been largely supported by animal models1,2. Since the publication of the first orthotopic rat ...

Ricerca

Medicina

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.

Procedure chirurgiche per un modello di Rat parziale trapianto di fegato con la ricostruzione arteriosa epatica

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

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 eterotopico addominale Cuore / Trapianti Single-polmone in una configurazione del volume-caricato

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.

Eterotopico cervicale trapianto cardiaco nei topi

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

Method of Direct Segmental Intra-hepatic Delivery Using a Rat Liver Hilar Clamp Model

Major hepatic surgery with inflow occlusion, and liver transplantation, necessitate a period of warm ischemia, and a period of reperfusion leading to ischemia/reperfusion (I/R) injury with myriad negative consequences. Potential I/R injury in marginal organs destined for liver transplantation contributes to the current donor shortage secondary to a decreased organ utilization rate. A significant need exists to explore hepatic I/R injury in order to mediate its impact on graft function in transplantation. Rat liver hilar clamp models are used to investigate the impact of different molecules on hepatic I/R injury. Depending on the model, these molecules have been delivered using inhalation, epidural infusion, intraperitoneal injection, intravenous administration or injection into the peripheral superior mesenteric vein. A rat liver hilar clamp model has been developed for use in studying the impact of pharmacologic molecules in ameliorating I/R injury. The described model for rat liver hilar clamp includes direct cannulation of the portal supply to the ischemic hepatic segment via a side branch of the portal vein, allowing for direct segmental hepatic delivery. Our approach is to induce ischemia in the left lateral and median lobes for 60 min, during which time the substance under study is infused. In this case, pegylated-superoxide dismutase (PEG-SOD), a free radical scavenger, is infused directly into the ischemic segment. This series of experiments demonstrates that infusion of PEG-SOD is protective against hepatic I/R injury. Advantages of this approach include direct injection of the molecule into the ischemic segment with consequent decrease in volume of distribution and reduction in systemic side effects.

A unique rat liver hilar clamp model was developed for studying the impact of pharmacologic molecules in ameliorating ischemia-reperfusion injury. This model includes direct cannulation of the portal supply to the ischemic liver segment via a branch of the portal vein, allowing for direct hepatic delivery.

Metodo di consegna diretta segmentale intra-epatico utilizzando un modello di fegato di ratto Hilar morsetto

Major hepatic surgery with inflow occlusion, and liver transplantation, necessitate a period of warm ischemia, and a period of reperfusion leading to ...

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.

Modello del ratto della partizione del fegato associando i e legatura della vena portale per procedura di messa in scena epatectomia (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.

Ri-Arterialized trapianto parziale di fegato del ratto con un in vivo vaso-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. ...

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.

Emodinamico invasivo caratterizzazione della sindrome Portal-ipertesi in ratti cirrotici

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.

Generazione di un modello di ratto di insufficienza epatica acuta combinando il 70% di epatitectomia parziale e acetaminofene

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

Reduced Complications after Arterial Reconnection in a Rat Model of Orthotopic Liver Transplantation

The rat orthotopic liver transplantation (OLT) model is a powerful tool to study acute and chronic rejection. However, it is not a complete representation of human liver transplantation due to the absence of arterial reconnection. Described here is a modified transplantation procedure that includes the incorporation of hepatic artery (HA) reconnection, leading to a marked improvement in transplant outcomes. With a mean anhepatic time of 12 min and 14 s, HA reconnection results in improved perfusion of the transplanted liver and an increase in long-term recipient survival from 37.5% to 88.2%. This protocol includes the use of 3D-printed cuffs and holders to connect the portal vein and infrahepatic inferior vena cava. It can be implemented for studying multiple aspects of liver transplantation, from immune response and infection to technical aspects of the procedure. By incorporating a simple and practical method for arterial reconnection using a microvascular technique, this modified rat OLT protocol closely mimics aspects of human liver transplantation and will serve as a valuable and clinically relevant research model.

The goal of this study is to modify the rat orthotopic liver transplant model to better represent human liver transplantation and improve recipient survival. The presented method reestablishes hepatic arterial inflow by connecting the donor liver&#39;s common hepatic artery to the recipient liver&#39;s proper hepatic artery.

Complicazioni ridotte dopo la riconnessione arteriosa in un modello di ratto di trapianto di fegato ortotopico

The rat orthotopic liver transplantation (OLT) model is a powerful tool to study acute and chronic rejection. However, it is not a complete representation ...