Name: percutaneous hepatic perfusion (php) with melphalan as a treatment for unresectable metastases confined to the liver
Uploaded: 2016-07-31T14:00:00
Description: Watch this Scientific Journal Video about Percutaneous Hepatic Perfusion PHP with Melphalan as a Treatment for Unresectable Metastases Confined to the Liver at JoVE.com

The authors have no acknowledgements. 
The phase II study is financially supported by Delcath Systems.

Note: After a patient met all inclusion criteria and was carefully evaluated by a medical oncologist, surgeon and anaesthesiologist, a patient was included in the study. All patients provided written informed consent. Both clinical studies were approved by the Local Medical Ethics Committee of the Leiden University Medical Centre and are performed in accordance with the ethical standards of the Helsinki Declaration.
1. Pre-procedural Angiography by Interventional Radiologist
<ol>
	<li>In pre...

<ol>
	<li>Jovanovic, 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=Ocular+melanoma:+an+overview+of+the+current+status.">Ocular melanoma: an overview of the current status.</a> Int J Clin Exp Pathol. 6 (7), 1230-1244 (2013).</li><li>Taylor, I., Bennett, R., Sherriff, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+blood+supply+of+colorectal+liver+metastases.">The blood supply of colorectal liver metastases.</a> Br J Cancer. 38 (6), 749-756 (1978).</li><li>van de Velde, C. 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=A+successful+technique+of+in+vivo+isolated+liver+perfusion+in+pigs.">A successful technique of in vivo isolated liver perfusion in pigs.</a> J Surg Res. 41 (6), 593-599 (1986).</li><li>Vahrmeijer, A. L., Der Eb, M. M. V. a. n., Van Dierendonck, J. H., Kuppen, P. J., Van De Velde, C. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Delivery+of+anticancer+drugs+via+isolated+hepatic+perfusion:+a+promising+strategy+in+the+treatment+of+irresectable+liver+metastases?.">Delivery of anticancer drugs via isolated hepatic perfusion: a promising strategy in the treatment of irresectable liver metastases?.</a> Semin Surg Oncol. 14 (3), 262-268 (1998).</li><li>Vahrmeijer, A. L., van de Velde, C. J., Hartgrink, H. H., Tollenaar, R. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Treatment+of+melanoma+metastases+confined+to+the+liver+and+future+perspectives.">Treatment of melanoma metastases confined to the liver and future perspectives.</a> Dig Surg. 25 (6), 467-472 (2008).</li><li>Ben-Shabat, I., 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=Isolated+hepatic+perfusion+as+a+treatment+for+liver+metastases+of+uveal+melanoma.">Isolated hepatic perfusion as a treatment for liver metastases of uveal melanoma.</a> J Vis Exp. (95), e52490 (2015).</li><li>Vahrmeijer, A. L., 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=Increased+local+cytostatic+drug+exposure+by+isolated+hepatic+perfusion:+a+phase+I+clinical+and+pharmacologic+evaluation+of+treatment+with+high+dose+melphalan+in+patients+with+colorectal+cancer+confined+to+the+liver.">Increased local cytostatic drug exposure by isolated hepatic perfusion: a phase I clinical and pharmacologic evaluation of treatment with high dose melphalan in patients with colorectal cancer confined to the liver.</a> Br J Cancer. 82 (9), 1539-1546 (2000).</li><li>Rothbarth, 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=Isolated+hepatic+perfusion+with+high-dose+melphalan+for+the+treatment+of+colorectal+metastasis+confined+to+the+liver.">Isolated hepatic perfusion with high-dose melphalan for the treatment of colorectal metastasis confined to the liver.</a> Br J Surg. 90 (11), 1391-1397 (2003).</li><li>van Iersel, L. B., 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=Isolated+hepatic+perfusion+with+200+mg+melphalan+for+advanced+noncolorectal+liver+metastases.">Isolated hepatic perfusion with 200 mg melphalan for advanced noncolorectal liver metastases.</a> Ann Surg Oncol. 15 (7), 1891-1898 (2008).</li><li>van Iersel, L. B., 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=Isolated+hepatic+melphalan+perfusion+of+colorectal+liver+metastases:+outcome+and+prognostic+factors+in+154+patients.">Isolated hepatic melphalan perfusion of colorectal liver metastases: outcome and prognostic factors in 154 patients.</a> Ann Oncol. 19 (6), 1127-1134 (2008).</li><li>Alexander, H. R., 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=A+phase+I-II+study+of+isolated+hepatic+perfusion+using+melphalan+with+or+without+tumor+necrosis+factor+for+patients+with+ocular+melanoma+metastatic+to+liver.">A phase I-II study of isolated hepatic perfusion using melphalan with or without tumor necrosis factor for patients with ocular melanoma metastatic to liver.</a> Clin Cancer Res. 6 (8), 3062-3070 (2000).</li><li>Olofsson, R., 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=Isolated+hepatic+perfusion+for+ocular+melanoma+metastasis:+registry+data+suggests+a+survival+benefit.">Isolated hepatic perfusion for ocular melanoma metastasis: registry data suggests a survival benefit.</a> Ann Surg Oncol. 21 (2), 466-472 (2014).</li><li>Curley, S. A., 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=Increased+doxorubicin+levels+in+hepatic+tumors+with+reduced+systemic+drug+exposure+achieved+with+complete+hepatic+venous+isolation+and+extracorporeal+chemofiltration.">Increased doxorubicin levels in hepatic tumors with reduced systemic drug exposure achieved with complete hepatic venous isolation and extracorporeal chemofiltration.</a> Cancer Chemother Pharmacol. 33 (3), 251-257 (1993).</li><li>Ravikumar, T. S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Percutaneous+hepatic+vein+isolation+and+high-dose+hepatic+arterial+infusion+chemotherapy+for+unresectable+liver+tumors.">Percutaneous hepatic vein isolation and high-dose hepatic arterial infusion chemotherapy for unresectable liver tumors.</a> J Clin Oncol. 12 (12), 2723-2736 (1994).</li><li>Lillemoe, H. A., Alexander, H. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Current+Status+of+Percutaneous+Hepatic+Perfusion+as+Regional+Treatment+for+Patients+with+Unresectable+Hepatic+Metastases:+A+Review.">Current Status of Percutaneous Hepatic Perfusion as Regional Treatment for Patients with Unresectable Hepatic Metastases: A Review.</a> Am Oncology and Hematology Rev. (15-23), (2014).</li><li>Pingpank, J. F., 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=Phase+I+study+of+hepatic+arterial+melphalan+infusion+and+hepatic+venous+hemofiltration+using+percutaneously+placed+catheters+in+patients+with+unresectable+hepatic+malignancies.">Phase I study of hepatic arterial melphalan infusion and hepatic venous hemofiltration using percutaneously placed catheters in patients with unresectable hepatic malignancies.</a> J Clin Oncol. 23 (15), 3465-3474 (2005).</li><li>Fitzpatrick, M., 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=Use+of+partial+venovenous+cardiopulmonary+bypass+in+percutaneous+hepatic+perfusion+for+patients+with+diffuse,+isolated+liver+metastases:+a+case+series.">Use of partial venovenous cardiopulmonary bypass in percutaneous hepatic perfusion for patients with diffuse, isolated liver metastases: a case series.</a> J Cardiothorac Vasc Anesth. 28 (3), 647-651 (2014).</li><li>Forster, M. R., 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=Chemosaturation+with+percutaneous+hepatic+perfusion+for+unresectable+metastatic+melanoma+or+sarcoma+to+the+liver:+a+single+institution+experience.">Chemosaturation with percutaneous hepatic perfusion for unresectable metastatic melanoma or sarcoma to the liver: a single institution experience.</a> J Surg Oncol. 109 (5), 434-439 (2014).</li><li>Deneve, J. L., 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=Chemosaturation+with+percutaneous+hepatic+perfusion+for+unresectable+isolated+hepatic+metastases+from+sarcoma.">Chemosaturation with percutaneous hepatic perfusion for unresectable isolated hepatic metastases from sarcoma.</a> Cardiovasc Intervent Radiol. 35 (6), 1480-1487 (2012).</li><li>Kandarpa, K., Machan, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Handbook of Interventional Radiologic Procedures. , 269-270 (2011).</li><li>Hofmann, H., 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=Unresectable+isolated+hepatic+metastases+from+solid+pseudopapillary+neoplasm+of+the+pancreas:+a+case+report+of+chemosaturation+with+high-dose+melphalan.">Unresectable isolated hepatic metastases from solid pseudopapillary neoplasm of the pancreas: a case report of chemosaturation with high-dose melphalan.</a> Pancreatology. 14 (6), 546-549 (2014).</li><li>Miao, N., 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=Percutaneous+hepatic+perfusion+in+patients+with+metastatic+liver+cancer:+anesthetic,+hemodynamic,+and+metabolic+considerations.">Percutaneous hepatic perfusion in patients with metastatic liver cancer: anesthetic, hemodynamic, and metabolic considerations.</a> Ann Surg Oncol. 15 (3), 815-823 (2008).</li><li>Vogl, T. 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=Chemosaturation+with+percutaneous+hepatic+perfusions+of+melphalan+for+hepatic+metastases:+experience+from+two+European+centers.">Chemosaturation with percutaneous hepatic perfusions of melphalan for hepatic metastases: experience from two European centers.</a> Rofo. 186 (10), 937-944 (2014).</li><li>Pingpank, J. F., 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=A+phase+III+random+assignment+trial+comparing+percutaneous+hepatic+perfusion+with+melphalan+(PHP-mel)+to+standard+of+care+for+patients+with+hepatic+metastases+from+metastatic+ocular+or+cutaneous+melanoma.">A phase III random assignment trial comparing percutaneous hepatic perfusion with melphalan (PHP-mel) to standard of care for patients with hepatic metastases from metastatic ocular or cutaneous melanoma.</a> J Clin Oncol. 28, 18s (2010).</li><li>Rothbarth, J., Vahrmeijer, A. L., Mulder, G. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Modulation+of+cytostatic+efficacy+of+melphalan+by+glutathione:+mechanisms+and+efficacy.">Modulation of cytostatic efficacy of melphalan by glutathione: mechanisms and efficacy.</a> Chem Biol Interact. 140 (2), 93-107 (2002).</li><li>Bartlett, D. L., Libutti, S. K., Figg, W. D., Fraker, D. L., Alexander, H. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Isolated+hepatic+perfusion+for+unresectable+hepatic+metastases+from+colorectal+cancer.">Isolated hepatic perfusion for unresectable hepatic metastases from colorectal cancer.</a> Surgery. 129 (2), 176-187 (2001).</li><li>van Iersel, L. B., 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=Isolated+hepatic+perfusion+with+oxaliplatin+combined+with+100+mg+melphalan+in+patients+with+metastases+confined+to+the+liver:+A+phase+I+study.">Isolated hepatic perfusion with oxaliplatin combined with 100 mg melphalan in patients with metastases confined to the liver: A phase I study.</a> Eur J Surg Oncol. , (2014).</li><li>Rothbarth, J., Tollenaar, R. A., van de Velde, C. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Recent+trends+and+future+perspectives+in+isolated+hepatic+perfusion+in+the+treatment+of+liver+tumors.">Recent trends and future perspectives in isolated hepatic perfusion in the treatment of liver tumors.</a> Expert Rev Anticancer Ther. 6 (4), 553-565 (2006).</li><li>van Etten, B., de Wilt, J. H., Brunstein, F., Eggermont, A. M., Verhoef, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Isolated+hypoxic+hepatic+perfusion+with+melphalan+in+patients+with+irresectable+ocular+melanoma+metastases.">Isolated hypoxic hepatic perfusion with melphalan in patients with irresectable ocular melanoma metastases.</a> Eur J Surg Oncol. 35 (5), 539-545 (2009).</li><li>van Iersel, L. B., 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=Management+of+isolated+nonresectable+liver+metastases+in+colorectal+cancer+patients:+a+case-control+study+of+isolated+hepatic+perfusion+with+melphalan+versus+systemic+chemotherapy.">Management of isolated nonresectable liver metastases in colorectal cancer patients: a case-control study of isolated hepatic perfusion with melphalan versus systemic chemotherapy.</a> Ann Oncol. 21 (8), 1662-1667 (2010).</li></ol>

Patients with unresectable liver metastases can be treated with systemic therapy. However, for patients with metastatic uveal melanoma, no standard systemic therapy is available and immunotherapy or targeted therapy have not yet been able to show improved survival. Isolated hepatic perfusion has been shown to be an effective treatment for patients with unresectable uveal melanoma metastases confined to the liver.9,28 For colorectal cancer metastases more therapeutic systemic options are available, but some pat...

Resection of malignant liver tumors is the first choice of treatment for both primary and secondary hepatic malignancies. However, a large proportion of patients are not candidates for surgery because of extended disease or location of the metastases. For patients with unresectable metastases from colorectal carcinoma, systemic therapy is often the preferred treatment. Hepatic metastases from uveal melanoma are often small and diffusely spread throughout the liver. No standard systemic therapy is available for this group of patients. Local therapy can be an alternative to systemic treatment, in case the metastases are confined to the liver.
Because of the specific vascular anatomy of the liver, this organ can be isolated from the systemic circulation. This allows perfusion of the liver with high dose chemotherapy (IHP, isolated hepatic perfusion). Besides, liver malignancies have a dominant or exclusive vascular supply from the hepatic artery, whereas 70-80% of the supply of the non-tumorous liver parenchyma is derived from the portal vein.1,2 This technique was developed over twenty years ago, to treat patients with unresectable metastases from various primary origins.3,4 Especially, uveal melanoma patients with metastases in the liver may be candidates for IHP because the metastases are often small and spread throughout the entire liver, and at present no standard systemic therapy is available.5,6
The principle of IHP is to temporarily isolate the liver from the systemic circulation and perfuse the organ with a high dose of chemotherapy, leading to high local drug exposure with limited systemic side effects.7 This high dose of chemotherapy would be toxic and lead to complications when administered systemically. The majority of IHP studies were performed with melphalan, and have investigated treatment of hepatic metastasis from colorectal cancer patients, as well as patients with uveal melanoma metastases.8,9 Several studies of IHP during open surgery suggest that this treatment might be effective: 50-59% tumor response rates (partial and complete response) for the treatment of colorectal cancer and a 68% tumor response rate for patients with metastatic uveal melanoma have been reported.8,10,11,12 Despite these treatment results, this procedure never gained wide acceptance, because of the complexity of the procedure, the duration of hospital stay and the associated morbidity and mortality.
Percutaneous hepatic perfusion (PHP) offers a minimal invasive alternative to IHP and was first demonstrated in a porcine model in 1993 using doxorubicin13 and the first in human trial was performed by Ravikumar et al. in 1994.14 Due to lack of evidence of efficacy, the technique was largely abandoned until the early 2000&#39;s when it was re-evaluated in the National Cancer Institute (NCI) in the United States.15 During PHP, a catheter is placed percutaneous into the proper hepatic artery via the femoral artery to infuse the chemotherapeutic agent. A second catheter is placed in the inferior caval vein via the femoral vein to aspirate the hepatic chemosaturated outflow (see the PHP circuit in Figure 1). The isolation aspiration catheter placed in the caval vein is a double balloon catheter, prohibiting leakage into the systemic circulation. The aspirated chemosaturated blood is filtered by a double charcoal filter and returned to the patient by a third catheter placed in the internal jugular vein. The patient is admitted in the hospital with a length of stay of ~3 days. The PHP procedure is performed in an angiography room under general anaesthesia by a well-trained multidisciplinary team consisting of a dedicated interventional radiologist, anaesthesiologist and an extracorporeal perfusionist. A surgical oncologist and medical oncologist are also members of this multidisciplinary team, and especially focus on informing the patient, patient selection and post-operative care.
<img alt="Figure 1" src="/files/ftp_upload/53795/53795fig1.jpg" /> 
Figure 1. Schematic image of the PHP circuit. This figure displays the set-up of the PHP circuit. It shows an isolated hepatic perfusion circuit with extra-corporeal bypass line. <a href="https://www.jove.com/files/ftp_upload/53795/53795fig1large.jpg" target="_blank">Please click here to view a larger version of this figure.</a>
This minimal invasive procedure is associated with less operative morbidity and can be repeated several times (at least up to 4 times). Besides, it only takes approximately 3 to 4 hr&#160;and patient recovery is fast. The advantage of PHP is the fact that all sizes of metastases can be treated, and micro metastases are being treated as well. Also the location of the metastases, close to vascular structures and bile ducts, is not a contraindication for PHP. Initial studies were performed with the 1st generation filter, with a 77% (mean) filter extraction efficiency.16 Recently, the results of a phase III trial were published by Hughes et al. showing a significant improvement of hepatic progression free survival in uveal melanoma patients with hepatic metastases treated with PHP compared to best alternative care.17
Since April 2012 a 2nd generation filter is available. In pre-clinical studies the 2nd generation filter is extracting 98% of melphalan. Several studies and case series investigating PHP for multiple indications have been published, but apart from the recent publishing phase III trial, survival has not extensively been analysed.16,18,19,20 In the present investigation, we focus on the interventional radiology procedure, as well as the anaesthetic management and the extra corporeal circulation that is used during this procedure in order to facilitate the use of this treatment in other medical centers.

<table border="0" cellpadding="0" cellspacing="0" width="1115">
	<tbody>
		<tr height="42">
			<td height="42" style="height:42px;width:411px;">Delcath 2nd Generation Hepatic CHEMOSAT Delivery System&#160;</td>
			<td style="width:357px;">Delcath Systems Inc., New York, New York, USA</td>
			<td style="width:180px;">602001 and 602002</td>
			<td style="width:167px;">Item no. 1</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Isofuse Isolation Aspiration Catheter (Double Balloon Catheter)</td>
			<td>Delcath Systems Inc., New York, New York, USA</td>
			<td></td>
			<td rowspan="2">Item no. 1a</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">10F Venous Return Catheter</td>
			<td>Delcath Systems Inc., New York, New York, USA</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Hemofiltration Cartridges</td>
			<td>Delcath Systems Inc., New York, New York, USA</td>
			<td></td>
			<td rowspan="2">Item no. 1b</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:411px;">Circuit components</td>
			<td>Delcath Systems Inc., New York, New York, USA</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Level-1 rapid fluid management system</td>
			<td style="width:357px;">Smiths Medical</td>
			<td style="width:180px;"></td>
			<td>Item no. 2</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;">22G arterial line&#160;</td>
			<td style="width:357px;">Arrow International Inc. /&#160; Teleflex Inc.,Dublin, Ireland</td>
			<td style="width:180px;"></td>
			<td>Item no. 3</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Vigileo, Monitor</td>
			<td>Edwards Lifesciences Corp, Irvine, California, USA</td>
			<td style="width:180px;"></td>
			<td>Item no. 4</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">7.5F Triple lumen intravenous catheter, 20 cm&#160;</td>
			<td>Vygon, Valkenswaard, Nederland</td>
			<td></td>
			<td>Item no. 5</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">5F sheath&#160;</td>
			<td style="width:357px;"></td>
			<td style="width:180px;"></td>
			<td>Item no. 6</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">2.7 Progreat microcatheter&#160;</td>
			<td>Terumo, Tokyo, Japan</td>
			<td>MC-PP27131</td>
			<td>Item no. 7</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">18F sheath&#160;</td>
			<td style="width:357px;"></td>
			<td style="width:180px;"></td>
			<td>Item no. 8</td>
		</tr>
		<tr height="84">
			<td height="84" style="height:84px;">Auto-injector Medrad Mark V ProVis</td>
			<td>Bayer, Indianola, Pennsylvania, USA</td>
			<td style="width:180px;">Not available anymore, replaced by&#160;Medrad&#160;Mark 7 Arterion&#160;Injection System</td>
			<td>Item no. 9</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:411px;">Melphalan</td>
			<td>Alkeran, Aspen Pharmacare, Dublin, Ireland</td>
			<td style="width:180px;"></td>
			<td>Item no. 10</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:411px;">Heparin LEO, 5000 IE/ml</td>
			<td style="width:357px;">LEO Pharma AB, Denmark</td>
			<td style="width:180px;"></td>
			<td>Item no. 11</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:411px;">Centrifugal pump</td>
			<td style="width:357px;">Medtronic, Minneapolis, Minnesota, VS</td>
			<td style="width:180px;"></td>
			<td>Item no. 12</td>
		</tr>
		<tr height="21">
			<td colspan="2" height="21" style="height:21px;">Voluven colloid solution (6% hydroxyethyl starch 130/0.4 in 0.9% sodium chloride injection)</td>
			<td style="width:180px;"></td>
			<td>Item no. 13</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:411px;">Iopromide 300, Ultravist</td>
			<td style="width:357px;">Bayer, Indianola, Pennsylvania, USA</td>
			<td style="width:180px;"></td>
			<td>Item no. 14</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:411px;">Detachable coil, Interlock</td>
			<td style="width:357px;">Boston Scientific, Marlborough, Massachusetts, USA</td>
			<td style="width:180px;"></td>
			<td>Item no. 15</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:411px;">Vascular plug, Amplatzer 4</td>
			<td style="width:357px;">St. Jude Medical, St Paul,Minnesota, USA</td>
			<td style="width:180px;"></td>
			<td>Item no. 16</td>
		</tr>
	</tbody>
</table>

percutaneous hepatic perfusion (php) with melphalan as a treatment for unresectable metastases confined to the liver

Unresectable liver metastases of colorectal cancer can be treated with systemic chemotherapy, aiming to limit the disease, extend survival or turn unresectable metastases into resectable ones. Some patients however, suffer from side effects or progression under systemic treatment. For patients with metastasized uveal melanoma there are no standard systemic therapy options. For patients without extrahepatic disease, isolated liver perfusion (IHP) may enable local disease control with limited systemic side effects. Previously, this was performed during open surgery with satisfying results, but morbidity and mortality related to the open procedure, prohibited a widespread application. Therefore, percutaneous hepatic perfusion (PHP) with simultaneous chemofiltration was developed. Besides decreasing morbidity and mortality, this procedure can be repeated, hopefully leading to a higher response rate and improved survival (by local control of disease). During PHP, catheters are placed in the proper hepatic artery, to infuse the chemotherapeutic agent, and in the inferior caval vein to aspirate the chemosaturated blood returning through the hepatic veins. The caval vein catheter is a double balloon catheter that prohibits leakage into the systemic circulation. The blood returning from the hepatic veins is aspirated through the catheter fenestrations and then perfused through an extra-corporeal filtration system. After filtration, the blood is returned to the patient by a third catheter in the right internal jugular vein. During PHP a high dose of melphalan is infused into the liver, which is toxic and would lead to life threatening complications when administered systemically. Because of the significant hemodynamic instability resulting from the combination of caval vein occlusion and chemofiltration, hemodynamic monitoring and hemodynamic support is of paramount importance during this complex procedure.

Knowledge about PHP is based on small phase I and II trials and case series and a recent larger phase trial III; an overview of published results is shown in Table 1. One paper discusses the anaesthesiology procedure, hemodynamic and metabolic aspects of the treatment. However, no survival data were reported.22 Three larger trials that were reported, included metastatic liver disease from different primary tumors and the results are therefore difficult to inter...

Watch this Scientific Journal Video about Percutaneous Hepatic Perfusion PHP with Melphalan as a Treatment for Unresectable Metastases Confined to the Liver at JoVE.com

Percutaneous Hepatic Perfusion PHP with Melphalan as a Treatment for Unresectable Metastases Confined to the Liver

In this manuscript, we describe percutaneous isolated hepatic perfusion with simultaneous chemofiltration as treatment for unresectable liver metastases. This procedure is performed under general anaesthesia in the angiosuite by an experienced team, consisting of an interventional radiologist, a clinical perfusionist and anaesthesiologist.

Percutaneous Hepatic Perfusion (PHP) with Melphalan as a Treatment for Unresectable Metastases Confined to the Liver

Unresectable liver metastases of colorectal cancer can be treated with systemic chemotherapy, aiming to limit the disease, extend survival or turn ...

The overall goal of this procedure is to carry out Percutaneous Hepatic Perfusion to treat unresectable metastases confined to the liver. The main advantage of this technique is that it allows delivery of a high dose of chemotherapeutic agents to the liver with limited systemic exposure. It is minimally invasive, and therefore associated with low mobility and mortality rates.And finally, an advantage is that it can be repeated. Demonstrating the procedure will be Christian Martini, the anesthesiologist, and Fred Tijl, the perfusionist. In preparation for the PHP procedure, perform a pre-procedural angiography several days prior to PHP, according to the text protocol.On the day of the PHP procedure, have the extracorporeal profusionist, or ECP, collect the chemo saturation tubing pack, chemo filters, and the centrifugal pump as provided by the manufacturer. With carbon dioxide gas, flush the complete extracorporeal circuit. Then, use 0.9%sodium chloride and heparin to prime the circuit, including the filters.Allow six liters of heparinized 0.9%sodium chloride to rinse the extracorporeal system initially through the force of gravity. Then after the carbon filters are fully hydrated, use the centrifugal pump to apply more pressure to the filter to remove the remaining gas bubbles. After the anesthesiologist prepares the patient according to the text protocol, have the interventional radiologist or IR, use ultrasound to place a 7.5F triple lumen, 27 centimeter intravenous catheter in the left internal jugular vein, or IJV.Next, place a 10F venous return catheter in the right IJV, and insert a 5F sheathe into the left common femoral artery, or CFA. Using eight to 10F and 12 to 14F dilaters, perform a serial dilation of the right common femoral vein, or CFV, using the dilaters to adjust the insertion opening to the correct size. Then insert an 18F sheathe that will be used to introduce the double balloon catheter.Check the blood pressure of the patient. Following the administration of heparin and after starting norepinephrine and phenylephrine according to the text protocol, through the 5F sheathe in the left CFA, introduce the 2.7F micro catheter coaxially, and with angiography monitoring, insert it into the proper hepatic artery or lobar arteries. Attach filters and pump to the sheathe in the jugular vein BIJECP.Through the 18F sheathe and the right CFV, introduce the 16F double balloon catheter or isolation aspiration catheter, and via the angiography monitor, place it with the tip in the right atrium. Attach the extracorporeal filter system to the balloon catheter. Fill the extracorporeal system with the patient's blood.Next, start the centrifugal pump per the manufacturer's instructions. And after intravenous infusion of 0.5 liters of colloid solution, use a 50%saline, 50%contrast medium mixture to inflate the cranial balloon. Monitor the blood pressure by anesthesiologist and radiologist, for a decrease in blood pressure is likely after inflation of the cranial balloon.Then with the coddled balloon still deflated, slowly retract the double balloon catheter until the cranial balloon is at the junction of the right atrium and the inferior caval vein or ICV, just above the right hepatic vein. The balloon will now be wedge shaped. Using a solution of 90%saline and 10%contrast medium, inflate the coddled balloon until the lateral edges start to become effaced by the ICV.To use phonography to check the position of the balloons and to check for leaks, by hand, inject 20 milliliters of contrast through the fenestration port of the venous double balloon catheter. Check the position of the balloons by retrograde injection of contrasts through the fenetrated double balloon catheter. When sufficient flows and pressures have been reached according to the text protocol, open the filters one by one.Then, when a systolic blood pressure between 140 and 160 milliliters of mercury has been reached, close the bypass line. Attach the melphalan into the injector and load the pump. Be aware of a drop in blood pressure.After performing another venography to check positioning of the double balloon, perform an angiogram by injecting contrast through the micro catheter in the hepatic artery to rule out arterial spasm. When a steady flow rate is established, no leakage is present, and the patient's blood pressure is stable, use an auto-injector to administer 100 milliliters of three milligrams per kilogram of melphalan into the proper hepatic artery or sequentially into both lobar arteries at a rate of 0.4 ccs per second. Following the 30 minutes of chemotherapeutic infusion, continue hemofiltration for an additional 30 minute washout period.After complete infusion, a 30 minute washout time follows. After the washout period, slowly deflate the double cava balloons. Then empty the extracorporeal circuit as much as possible before stopping the extracorporeal perfusion.Remove the double balloon catheter and the infusion micro catheter, but leave the access sheathes in place until coagulation is normalized. Place a vascular closure device in the common femoral artery if desired, to achieve hemostasis. Finally, close and remove the bypass circuit, stabilize the patient, and carry out post-operative care according to the text protocol.Shown here is an overview of small case one and two trials, and a case series for PHP. The first manuscript was published in 1994 and 5-FU and doxorubin were used. Published overall response rates varied between 30 and 90%and survival data were not reported.A recent phase three trial comparing PHP to best alternative care, or BAC, for patients with hepatic metastases of uveal melanoma reports significant improved hepatic progression-free survival of seven months, compared to 1.6 months for the group that received BAC. Once mastered, this procedure can be performed in three to four hours if performed properly. When attempting percutaneous hepatic perfusion, patient selection is very important.Patients should be fit to undergo treatment and they should have liver-dominant or liver-only disease. Patients that are most likely to be the best candidates for percutaneous hepatic perfusion are patients with metastases from ocular melanoma. After watching this video, you should be able to understand how percutaneous hepatic perfusion is performed in a patient with liver tumors.Bare in mind that the procedure is best performed by a well trained and dedicated team of interventional radiologists, anesthesiologists, perfusionists, and technicians. Don't forget that significant hemodynamic perturbations may occur during the procedure and may require administration of high doses of sympathomimetics. Bone marrow depression is not uncommon and may be anticipated by administration of granulocyte colony-stimulating factor one to two days after the procedure.

percutaneous-hepatic-perfusion-php-with-melphalan-as-treatment-for

Research

JoVE Journal

Medicine

The Measurement and Treatment of Suppression in Amblyopia

Amblyopia, a developmental disorder of the visual cortex, is one of the leading causes of visual dysfunction in the working age population. Current estimates put the prevalence of amblyopia at approximately 1-3%1-3, the majority of cases being monocular2. Amblyopia is most frequently caused by ocular misalignment (strabismus), blur induced by unequal refractive error (anisometropia), and in some cases by form deprivation.
Although amblyopia is initially caused by abnormal visual input in infancy, once established, the visual deficit often remains when normal visual input has been restored using surgery and/or refractive correction. This is because amblyopia is the result of abnormal visual cortex development rather than a problem with the amblyopic eye itself4,5 . Amblyopia is characterized by both monocular and binocular deficits6,7 which include impaired visual acuity and poor or absent stereopsis respectively. The visual dysfunction in amblyopia is often associated with a strong suppression of the inputs from the amblyopic eye under binocular viewing conditions8. Recent work has indicated that suppression may play a central role in both the monocular and binocular deficits associated with amblyopia9,10 .
Current clinical tests for suppression tend to verify the presence or absence of suppression rather than giving a quantitative measurement of the degree of suppression. Here we describe a technique for measuring amblyopic suppression with a compact, portable device11,12 . The device consists of a laptop computer connected to a pair of virtual reality goggles. The novelty of the technique lies in the way we present visual stimuli to measure suppression. Stimuli are shown to the amblyopic eye at high contrast while the contrast of the stimuli shown to the non-amblyopic eye are varied. Patients perform a simple signal/noise task that allows for a precise measurement of the strength of excitatory binocular interactions. The contrast offset at which neither eye has a performance advantage is a measure of the "balance point" and is a direct measure of suppression. This technique has been validated psychophysically both in control13,14 and patient6,9,11 populations.
In addition to measuring suppression this technique also forms the basis of a novel form of treatment to decrease suppression over time and improve binocular and often monocular function in adult patients with amblyopia12,15,16 . This new treatment approach can be deployed either on the goggle system described above or on a specially modified iPod touch device15.

Amblyopia is a developmental disorder of the visual cortex that is often accompanied by strong suppression of one eye. We present a new technique for measuring and treating interocular suppression in patients with amblyopia that can be deployed using virtual reality goggles or a portable iPod Touch device.

Amblyopia, a developmental disorder of the visual cortex, is one of the leading causes of visual dysfunction in the working age population. Current ...

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.

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

Osmotic Drug Delivery to Ischemic Hindlimbs and Perfusion of Vasculature with Microfil for Micro-Computed Tomography Imaging

Preclinical research in animal models of peripheral arterial disease plays a vital role in testing the efficacy of therapeutic agents designed to stimulate microcirculation. The choice of delivery method for these agents is important because the route of administration profoundly affects the bioactivity and efficacy of these agents1,2. In this article, we demonstrate how to locally administer a substance in ischemic hindlimbs by using a catheterized osmotic pump. This pump can deliver a fixed volume of aqueous solution continuously for an allotted period of time. We also present our mouse model of unilateral hindlimb ischemia induced by ligation of the common femoral artery proximal to the origin of profunda femoris and epigastrica arteries in the left hindlimb. Lastly, we describe the in vivo cannulation and ligation of the infrarenal abdominal aorta and perfusion of the hindlimb vasculature with Microfil, a silicone radiopaque casting agent. Microfil can perfuse and fill the entire vascular bed (arterial and venous), and because we have ligated the major vascular conduit for exit, the agent can be retained in the vasculature for future ex vivo imaging with the use of small specimen micro-CT3.

We show here the in vivo insertion of an osmotic pump for constant local drug delivery and the creation of hindlimb ischemia in a mouse model. Moreover, the hindlimb vasculature is perfused with Microfil, a silicone radiopaque agent, to prepare for micro-computed tomography (micro-CT) imaging.

Preclinical research in animal models of peripheral arterial disease plays a vital role in testing the efficacy of therapeutic agents designed to ...

A Preclinical Murine Model of Hepatic Metastases

Numerous murine models have been developed to study human cancers and advance the understanding of cancer treatment and development. Here, a preclinical, murine pancreatic tumor model of hepatic metastases via a hemispleen injection of syngeneic murine pancreatic tumor cells is described. This model mimics many of the clinical conditions in patients with metastatic disease to the liver. Mice consistently develop metastases in the liver allowing for investigation of the metastatic process, experimental therapy testing, and tumor immunology research.

A preclinical, murine model of hepatic metastases performed via a hemispleen injection technique.

Numerous murine models have been developed to study human cancers and advance the understanding of cancer treatment and development. Here, a preclinical, ...

Isolated Hepatic Perfusion as a Treatment for Liver Metastases of Uveal Melanoma

Isolated hepatic perfusion (IHP) is a procedure where the liver is surgically isolated and perfused with a high concentration of the chemotherapeutic agent melphalan. Briefly, the procedure starts with the setup of a percutaneous veno-venous bypass from the femoral vein to the external jugular vein. Via a laparotomy, catheters are then inserted into the proper hepatic artery and the caval vein. The portal vein and the caval vein, both supra- and infrahepatically, are then clamped. The arterial and venous catheters are connected to a heart lung machine and the liver is perfused with melphalan (1 mg/kg body weight) for 60 min. This way it is possible to locally perfuse the liver with a high dose of a chemotherapeutic agent, without leakage to the systemic circulation.
In previous studies including patients with isolated liver metastases of uveal melanoma, an overall response rate of 33-100% and a median survival between 9 and 13 months, have been reported. The aim of this protocol is to give a clear description of how to perform the procedure and to discuss IHP as a treatment option for liver metastases of uveal melanoma.

Here, we present a protocol how to perform an isolated liver perfusion (IHP) with melphalan and we also discuss IHP as a treatment option for liver metastases of uveal melanoma.

Isolated hepatic perfusion (IHP) is a procedure where the liver is surgically isolated and perfused with a high concentration of the chemotherapeutic ...

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.

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

Treatment of Liver Metastases Using an Internal Target Volume Method for Stereotactic Body Radiotherapy

The prognosis of patients with metastatic cancers has improved in the past decades due to effective chemotherapy and oligometastatic surgery. For inoperable patients, local ablation therapies, such as stereotactic body radiotherapy (SBRT), can provide effective local tumor control with minimal toxicity. Because of its high precision and accuracy, SBRT delivers a higher radiation dose per fraction, is more effective, and targets smaller irradiation volumes than does conventional radiotherapy. In addition, steep dose gradients from target lesions to surrounding normal tissues are achieved using SBRT; thus, SBRT provides more effective tumor control and exhibits fewer side effects than conventional radiotherapy. The use of SBRT is prevalent for treating intracranial lesions (known as stereotactic radiosurgery); however, it is now also used for treating spinal and adrenal metastases. Because of advancements in image-guided assistance and respiratory motion management, several studies have investigated the use of SBRT for treating lung or liver tumors, which move as a patient breathes. The results of these studies have suggested that SBRT favorably controls tumors in the case of moving lesions.
Four-dimensional computed tomography (4D-CT) with an abdominal compressor (AC) is clinically convenient for effective respiratory motion management. Because this method is noninvasive and allows free breathing, its use reduces complications. Furthermore, patients consider this method convenient. Moreover, it is considered more efficient than other methods of respiratory motion management by physicians and therapists. The use of 4D-CT with an AC for treating pulmonary lesions has also been widely investigated, and the technique is gaining acceptance for treating hepatic lesions. However, the protocols for using 4D-CT with an AC for treating hepatic lesions are different from those used for treating pulmonary lesions. In this article, we describe a new protocol for SBRT with 4D-CT and an AC for treating liver metastases.

Stereotactic body radiotherapy (SBRT) requires rigorous accuracy and precision for delivering high radiation doses per fraction to small treatment volumes to improve tumor control and simultaneously reduce toxicity. Herein, we present a noninvasive and clinically convenient respiratory motion management protocol for SBRT for liver metastases.

The prognosis of patients with metastatic cancers has improved in the past decades due to effective chemotherapy and oligometastatic surgery. For ...

A Small Animal Model of Ex Vivo Normothermic Liver Perfusion

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

A Small Animal Model of Ex Vivo Normothermic Liver Perfusion

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

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

Measurement of the Hepatic Venous Pressure Gradient and Transjugular Liver Biopsy

Here we provide a detailed protocol describing the clinical procedure of hepatic venous pressure gradient (HVPG) measurement in patients with advanced chronic liver disease followed by an instruction for transjugular biopsy. Under local anesthesia and ultrasound guidance, a catheter introducer sheath is placed in the right internal jugular vein. Using&#160;fluoroscopic guidance, a balloon catheter is advanced into the inferior vena cava (IVC) and inserted into a large hepatic vein. Correct and sufficient wedge position of the catheter is ensured by injecting contrast media while the balloon is blocking the outflow of the cannulated hepatic vein. After calibrating the external pressure transducer, continuous pressure recordings are obtained with triplicate recordings of the wedged hepatic venous pressure (WHVP) and free hepatic venous pressure (FHVP). The difference between FHVP and WHVP is referred to as HVPG, with values &#8805;10 mm Hg indicating clinically significant portal hypertension (CSPH). Before removing the catheter, pressure readings obtained in the IVC at the same level, as well as the right atrial pressure are recorded.
Finally, a transjugular liver biopsy can be obtained via the same vascular route. Different systems are available; however, core biopsy needles are preferred over aspiration needles, especially for cirrhotic livers. Again, under fluoroscopic guidance a biopsy needle introducer sheath is advanced into an hepatic vein. Next, the transjugular biopsy needle is gently advanced through the introducer sheath: (i) in case of aspiration biopsy, the needle is advanced into the liver parenchyma under aspiration and then removed quickly, or (ii) in case of a core biopsy, the cutting-mechanism is triggered inside the parenchyma. Several separate passages can be safely performed to obtain sufficient liver specimens via transjugular biopsy. In experienced hands, the combination of these procedures takes about 30-45 min.

Here, we present a protocol for measurement of hepatic venous pressure gradient (HVPG),the gold standard to diagnose clinically significant portal hypertension. Moreover, we describe how to perform a transjugular liver biopsy within the same session.

Here we provide a detailed protocol describing the clinical procedure of hepatic venous pressure gradient (HVPG) measurement in patients with advanced ...

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

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

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

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