Name: isolated hepatic perfusion as a treatment for liver metastases of uveal melanoma
Uploaded: 2015-01-25T14:00:00.000+00:00
Duration: 9 min 52 s
Description: Watch this Scientific Journal Video about Isolated Hepatic Perfusion as a Treatment for Liver Metastases of Uveal Melanoma at JoVE.com

Lennart Wiman (photographer) is acknowledged for excellent video recording.

المرضى شريطة موافقة خطية مستنيرة وتمت الموافقة على الدراسة من قبل مجلس المراجعة الأخلاقية الإقليمي في جامعة غوتنبرغ. 1. التخدير <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> إعداد كافة رصد (ECG، ضغط الدم صفعة) 17. </li><li style=";text-align:right;direction:rtl"> إدراج القسطرة فوق الجافية في أسفل إلى منتصف مستوى الصدري 17. </li><li style=";text-align:right;direction:rtl"> لحث التخدير مع البروبوفول 2 مغ / كغ، 150 ميكروغرام من الفنتانيل، وRocuronium 0.6 ملغ / كغ من خلال القسطرة الوريدية الطرفية إدراجها سابقا. </li><li style=";text-align:right;direction:rtl"> Preoxygenate وتدخله المريض. الحفاظ على التخدير مع سيفوفلوران والأكسجين والهواء 17. </li><li style=";text-align:right;direction:rtl"> اضافة الى وجود تجويف مزدوج القسطرة الوريدية المركزية وقسطرة الشرايين. مراقبة CVP والشرايين ضغط 17. </li></ol> 2. عزل الكبد والقسطرة السفن <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> إدارة IE 100 / كغ من الهيبارين وإعطاء العلاج الوقائي بالمضادات الحيوية (على سبيل المثال، 160 ملغ من ميثوبريم و 800 ملغ من سلفاميثوكسازول). </li><li style=";text-align:right;direction:rtl"> باستخدام طريقة سيلدينغر عن طريق الجلد، ووضع 17 الاب قسطرة في الوريد الفخذي و15 الاب قسطرة في الوريد الوداجي الخارجي 18. التحقق من مواقف القسطرة عن طريق التنظير. </li><li style=";text-align:right;direction:rtl"> إجراء عملية فتح البطن عن طريق شق على شكل حرف L من عملية الخنجري أسفل خط الوسط ومن ثم امتدت إلى شق تحت الضلع الأيمن (19). </li><li style=";text-align:right;direction:rtl"> تحقق من أن لا الانبثاث خارج الكبد موجودة في البطن من قبل التفتيش الكامل والجس. <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> إذا تم العثور على أي آفات مشبوهة، تحقق من الورم الخبيث من قبل القسم المجمدة. </li><li style=";text-align:right;direction:rtl"> إنهاء الإجراء إذا كان هناك دليل على مظهر خارج الكبد (تضخم الغدد periaortal، الانبثاث في البطن). لا يعتبر اعتلال العقد اللمفية في الكبد بورتا موانع. </li></ol></li><li style=";text-align:right;direction:rtl"> تعبئة الكبد بقسمة الرباط المنجلي، في الرباط الثلاثي الأيسر والأيمن والشريان التاجي الرباط 19. </li><li style=";text-align:right;direction:rtl"> تشريح الأجوفيةالوريد ابتداء من الأوردة الكلوية حتى الحجاب الحاجز مع ربط الأوردة رافد خلف الصفاق. </li><li style=";text-align:right;direction:rtl"> تحديد الوريد الكظري الأيمن ووضع حلقة سفينة حوله. </li><li style=";text-align:right;direction:rtl"> تحديد الوريد الغدد التناسلية الصحيح. تشريح ذلك لمدة 5-10 سم. وضع خياطة محفظة سلسلة حول مدخل الوريد الأجوف. <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> وضع رباط بشكل أقصى على المنوال الغدد التناسلية. باستخدام مقص، وفتح الغدد التناسلية الوريد 3-5 سم من الوريد الأجوف. </li><li style=";text-align:right;direction:rtl"> إدراج 18 الاب المقوى سلك القسطرة عن طريق الوريد الغدد التناسلية الصحيح ووضع طرف retrohepatically فوق الأوردة الكلوية. </li><li style=";text-align:right;direction:rtl"> وضع رباط حول الوريد الغدد التناسلية لتأمين موقف القسطرة. </li><li style=";text-align:right;direction:rtl"> تحقق من عودة الدم من خلال القسطرة. يعالج بالهيبارين القسطرة 18. </li></ol></li><li style=";text-align:right;direction:rtl"> تشريح الرباط الكبدي الإثناعشري وتصور الشريان الكبدي شيوعا، وهو الشريان معدي والشريان الكبدي السليم. <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> سفينة مكان حلقات حول آرتي الكبدي شيوعاراي والشريان gastrodoudenal. </li><li style=";text-align:right;direction:rtl"> تشريح الوريد البابي ووضع حلقة سفينة حوله. </li><li style=";text-align:right;direction:rtl"> عزل بقية الرباط تحتوي على القناة الصفراوية المشتركة، وتشمل كل هذا في حلقة السفينة. </li></ol></li><li style=";text-align:right;direction:rtl"> رفع الشريان معدي مع حلقة السفينة. بعد انقطاع تدفق الشرايين، وجعل شق باستخدام المقص. <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> إدراج 8-12 الاب القسطرة مع طرفها في الشريان الكبدي السليم. تأمين موقف مع رباط حول الشريان معدي. </li><li style=";text-align:right;direction:rtl"> تحقق عودة الدم في القسطرة ثم يعالج بالهيبارين عليه 18. </li></ol></li><li style=";text-align:right;direction:rtl"> رئيس نظام الارواء مع وحدة واحدة من الكريات الحمراء معبأة، و 100 مل من الزلال (50 جم / لتر)، و 100 مل من Tribonat (بيكربونات الصوديوم)، 2500 وحدة من الهيبارين (5،000 IE / مل) و 250 مل من بلورانيات. </li><li style=";text-align:right;direction:rtl"> توصيل الشرايين والقسطرة الوريد الأجوف إلى نظام نضح تتألف من آلة الرئة القلب مجهزة بمضخة الأسطوانة وسخان رogether مع الخزان، مكساج وأنابيب أنظمة الوريدية من النوع القابل للتصرف. </li><li style=";text-align:right;direction:rtl"> ربط القسطرة الوداجي الفخذ والخارجية إلى مضخة تجاوز venovenous تتكون من مضخة طرد مركزي، جنبا إلى جنب مع وحدة سخان والبدء الالتفافية. </li><li style=";text-align:right;direction:rtl"> استكمال عزل الكبد من خلال تشديد وقف النزف Rumel حول الوريد الأجوف infrahepatically فوق عروق الكلى وتحامل على الوريد الأجوف suprahepatically باستخدام الأوعية الدموية المشبك Klintmalm. <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> تشديد وقف النزف Rumel حول القناة الصفراوية المشتركة، الوريد البابي والوريد الكظري الأيمن والمشبك أخيرا الشريان الكبدي باستخدام المشبك الأوعية الدموية. </li></ol></li></ol> 3. الإرواء <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> بدء نضح يدويا ومراقبة مستوى الدم في الخزان الوريدي. ضبط معدل تدفق حتى يتم التوصل إلى مستوى ثابت للدولة، عادة ما يكون معدل التدفق بين 500-1،200 مل / دقيقة. </li><li style=";text-align:right;direction:rtl"> ضع تحقيقات الثرمستور إلى اليمين والكبد غادر الفصوص واحدة مباشرة فيالقسطرة الشريانية. </li><li style=";text-align:right;direction:rtl"> تسخين سائل الإرواء تهدف إلى درجة حرارة الكبد الهدف من 40 درجة مئوية. تبقي درجة الحرارة هذه في جميع أنحاء الارواء. </li><li style=";text-align:right;direction:rtl"> وضع مسبار التلألؤ على venovenous رئيس مضخة الالتفافية. </li><li style=";text-align:right;direction:rtl"> معايرة نظام رصد التسرب، عن طريق حقن جرعة صغيرة من 10 من mbq 99m TC-الزلال (1 مل) إلى الدوران الجهازي وتسجيل النشاط الأساسي. <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> حقن عشرة أضعاف الجرعة أعلى، و 100 من mbq 99m TC-الزلال (1 مل)، في وحدة نضح الكبدية. </li><li style=";text-align:right;direction:rtl"> تسجيل أي تسرب من نظام نضح إلى الدوران الجهازي حيث أن زيادة النشاط تقاس التحقيق التلألؤ. </li><li style=";text-align:right;direction:rtl"> تسجيل أي تسرب من جهاز الدورة الدموية للنظام نضح عن زيادة في حجم في الخزان الوريدي. </li></ol></li><li style=";text-align:right;direction:rtl"> عندما يتم تأسيس دولة ثابتة لمعدل التدفق، أي تسرب موجود ولقد وصلت درجة حرارة الكبد 40 درجة مئوية، وضخ ملفلان بجرعة1 ملغم / كغم من وزن الجسم في النظام نضح مقسمة إلى جرعتين تعطى 30 دقيقة على حدة. </li></ol> 4. إنهاء الإرواء وإزالة القسطرة <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> بعد 60 دقيقة من نضح، تفريغ سائل الإرواء من النظام وري الكبد مع 1،000 مل من بلوراني. </li><li style=";text-align:right;direction:rtl"> إضافة وحدة واحدة من الكريات الحمراء معبأة إلى وحدة التروية، ولبث في الكبد ومن ثم وقف نضح. </li><li style=";text-align:right;direction:rtl"> الافراج عن المشبك Klintmalm ووقف النزف Rumel من الوريد الأجوف. </li><li style=";text-align:right;direction:rtl"> الافراج عن وقف النزف Rumel من الرباط الكبدي الإثناعشري. </li><li style=";text-align:right;direction:rtl"> إزالة ربطة من الشريان معدي. إزالة القسطرة وligate السفينة. </li><li style=";text-align:right;direction:rtl"> الافراج عن المشبك من الشريان الكبدي. </li><li style=";text-align:right;direction:rtl"> إزالة القسطرة من الوريد الغدد التناسلية وligate السفينة. </li><li style=";text-align:right;direction:rtl"> وقف-VENO الوريدي ضخ الالتفافية وقطع الأنابيب. </li><li style=";text-align:right;direction:rtl"> إغلاق البطن مع خياطة فافية دون انقطاع ومن ثم لخياطة الجلد داخل الجلد. </li><li style=";text-align:right;direction:rtl"> إزالة جatheters في عروق الوريد الفخذي والخارجية والضغط المحلي لمدة 5 دقائق. </li></ol> 5. بعد العملية الجراحية العناية <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> ينزع الأنبوب المريض ونقل إلى وحدة العناية بعد الجراحة لمدة 24 ساعة. حافظ على مراقبة الضغط الوريدي المركزي وضغط الدم الشرياني. </li><li style=";text-align:right;direction:rtl"> نقل المريض إلى قسم الجراحة لنموذجي من 3-7 أيام من الرعاية اللاحقة للعمليات الجراحية. </li><li style=";text-align:right;direction:rtl"> تواصل مع منخفضة الوزن الجزيئي هيبارين (LMWH) بجرعة من 5،000 IE / يوم أي ما مجموعه 4 أسابيع. </li></ol>

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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> Annals of Surgical Oncology. 15 (7), 1891-1898 (2008).</li><li>Carvajal, R. D., 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=Effect+of+selumetinib+vs+chemotherapy+on+progression-free+survival+in+uveal+melanoma:+a+randomized+clinical+trial.">Effect of selumetinib vs chemotherapy on progression-free survival in uveal melanoma: a randomized clinical trial.</a> JAMA : the Journal Of The American Medical Association. 311 (23), 2397-2405 (2014).</li><li>Sharma, K. V., 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=Hepatic+arterial+chemoembolization+for+management+of+metastatic+melanoma.">Hepatic arterial chemoembolization for management of metastatic melanoma.</a> AJR. American Journal Of Roentgenology. 190 (1), 99-104 (2008).</li><li>Eldredge-Hindy, 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=Yttrium-90+Microsphere+Brachytherapy+for+Liver+Metastases+From+Uveal+Melanoma:+Clinical+Outcomes+and+the+Predictive+Value+of+Fluorodeoxyglucose+Positron+Emission+Tomography.">Yttrium-90 Microsphere Brachytherapy for Liver Metastases From Uveal Melanoma: Clinical Outcomes and the Predictive Value of Fluorodeoxyglucose Positron Emission Tomography.</a> American Journal Of Clinical Oncology. , (2014).</li><li>Etten, 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+hypoxic+hepatic+perfusion+with+orthograde+or+retrograde+flow+in+patients+with+irresectable+liver+metastases+using+percutaneous+balloon+catheter+techniques:+a+phase+I+and+II+study.">Isolated hypoxic hepatic perfusion with orthograde or retrograde flow in patients with irresectable liver metastases using percutaneous balloon catheter techniques: a phase I and II study.</a> Annals of Surgical Oncology. 11 (6), 598-605 (2004).</li><li>Ku, Y., 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=Extracorporeal+adriamycin-removal+following+hepatic+artery+infusion:+use+of+direct+hemoperfusion+combined+with+veno-venous+bypass.">Extracorporeal adriamycin-removal following hepatic artery infusion: use of direct hemoperfusion combined with veno-venous bypass.</a> Nihon Geka Gakkai Zasshi. 90 (10), 1758-1764 (1989).</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=Hepatic+arterial+infusion+chemotherapy+with+complete+hepatic+venous+isolation+and+extracorporeal+chemofiltration:+a+feasibility+study+of+a+novel+system.">Hepatic arterial infusion chemotherapy with complete hepatic venous isolation and extracorporeal chemofiltration: a feasibility study of a novel system.</a> Anti-Cancer Drugs. 2 (2), 175-183 (1991).</li><li>Beheshti, M. V., 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+isolated+liver+perfusion+for+treatment+of+hepatic+malignancy:+preliminary+report.">Percutaneous isolated liver perfusion for treatment of hepatic malignancy: preliminary report.</a> Journal of Vascular And Interventional Radiology : JVIR. 3 (3), 453-458 (1992).</li></ol>

The authors have nothing to disclose.

المرضى الذين يعانون من سرطان الجلد المنتشر العنبي يكون عدد قليل من الخيارات العلاجية. والعلاج الكيميائي 6، العلاج المناعي 23 و العلاج المستهدف 24 لم تكن قادرة على أن تظهر تحسن OS. للمرضى الذين يعانون القليلة الانبثاث الكبد، استئصال الكبد هو خيار 12.</s...

سرطان الجلد العنبي هو الورم الخبيث العين الأساسي الأكثر شيوعا في البالغين، مع نسبة لها تكون أعلى نسبة في السكان قوقازي 1. يتكون العلاج المحلية من استئصال أو البلاك المحلية العلاج الإشعاعي 2،3. بغض النظر عن العلاج المحلية، والمرض المنتشر تتطور في نهاية المطاف في حوالي 35-50٪ من المرضى 4،5. الكبد هو الموقع الأكثر شيوعا لالانبثاث وحوالي 50٪ من المرضى الحالي مع الانبثاث الكبد معزولة. ومتوسط ​​البقاء الإجمالي (OS) للمرضى الذين يعانون من الانبثاث الكبد هو 6-12 4 أشهر. تم استكشاف العديد من الاستراتيجيات المختلفة العلاج الشامل والإقليمية، ولكن معدلات البقاء على قيد الحياة لم تتحسن 2. العلاج مع العلاج الكيميائي النظامية، وكلاهما مادة واحدة (داكاربازين أو Temozolomide 6) أو مزيج من العلاجات (نظام بولد 7، جيمسيتابين مع treosulfan 8)، ويظهر معدلات الاستجابة أقل ثم10٪. وقد أظهرت الكبدي العلاج الكيميائي داخل الشريان (HIA) معدل أعلى قليلا العام استجابة للعلاج الكيميائي مقارنة النظامية، ولكن أي زيادة في بقاء الكلي 9. أظهر chemoembolization Transarterial (TACE) مع سيسبلاتين وكاربوبلاتين استجابة جزئية في 57٪ من المرضى الذين يعانون من البقاء على قيد الحياة وسيطة من 9 أشهر 10. وقد أظهرت العلاج الإشعاعي الداخلي الانتقائي (SIRT) استجابة جزئية في 62٪ من المرضى الذين يعانون من البقاء على قيد الحياة وسيطة من 7 أشهر 11. في دراسة عن استئصال الكبد، بما في ذلك 255 مريضا، تم التبليغ عن البقاء الإجمالي 14 شهرا. في 76 من المرضى حيث كان الاستئصال الكامل مجهريا ممكن، وبقاء 27 شهرا 12. نضح الكبدي معزولة (IHP) هو خيار العلاج الإقليمي، وضعت أصلا من قبل ريان وAusman. الأساس المنطقي وراء هذه التقنية هو عزل جراحيا ويروي الكبد مع جرعة عالية من العلاج الكيميائي وكيل مع تجنب النظامية لxicity 13. وبالإضافة إلى ذلك، ارتفاع الحرارة المتزامنة يتوسط لامتصاص زيادة من وكيل العلاج الكيميائي بسبب التغيرات في تدفق الدم وورم نفاذية الغشاء الخلوي 14. ويؤكد كاملة العزلة الأوعية الدموية مع تقنية التتبع المشعة باستخدام TC-99 وصفت المصل البشري الزلال 15. وقد أظهرت دراسة استعادية المرحلة الثانية فائدة البقاء على قيد الحياة المحتملة من 14 شهرا (26 مقابل 12 شهرا) للمرضى التي خضعت IHP مقارنة أطول الناجين مع الانبثاث الكبد سرطان الجلد عنبي في السويد 16. هنا نقدم تقنية IHP مع لمحة موجزة عن النتائج الحالية والآفاق المستقبلية.

<table><tbody><tr><td>Wire re-inforced catethers</td><td>Medtronic Inc, Minnesota, USA</td><td></td><td>12-18Fr</td></tr><tr><td>St&amp;ouml;ckert S5</td><td>Sorin Group, Mirandola, Italy</td><td></td><td>Heart-lung machine</td></tr><tr><td>Rotaflow</td><td>Maquet, Jostra Medizintechnik AG, Hirrlingen, Germany</td><td></td><td>Centrifugal pump</td></tr><tr><td>HU 35</td><td>Maquet cardiopulmonary AG, Hirrlingen, Germany</td><td></td><td>Heater unit</td></tr><tr><td>Flexbumin 200 g/l</td><td>Baxter Medical AB, Kista, Sweden</td><td></td><td></td></tr><tr><td>Tribonat</td><td>Fresenius Kabi AB, Uppsala, Sweden</td><td></td><td></td></tr><tr><td>Heparin LEO, 5000 IE/ml</td><td>LEO Pharma AB, Sweden</td><td></td><td></td></tr><tr><td>Ringer Acetate</td><td>Baxter Medical AB, Kista, Sweden</td><td></td><td></td></tr><tr><td>Vasculosis</td><td>Cis-Bio International, Gif-Sur-Yvette, France</td><td></td><td>99mTc-albumin</td></tr><tr><td>MedicView</td><td>SystemData AB, Gothenburg, Sweden</td><td></td><td>Leakage monitoring system</td></tr><tr><td>Alkeran</td><td>Aspen Europe GmbH, Bad Oldesloe, Germany</td><td></td><td>melphalan</td></tr></tbody></table>

isolated hepatic perfusion as a treatment for liver metastases of uveal melanoma

نضح الكبدي معزولة (IHP) هو الإجراء حيث يتم عزل الكبد جراحيا ومع perfused على نسبة عالية من وكيل ملفلان العلاج الكيميائي. لفترة وجيزة، يبدأ الإجراء مع الإعداد لتجاوز-VENO الوريدي عن طريق الجلد من الوريد الفخذي إلى الوريد الوداجي الخارجي. عن طريق فتح البطن، ثم يتم إدخال القسطرة في الشريان الكبدي السليم والوريد الأجوف. الوريد البابي والوريد الأجوف، سواء supra- وinfrahepatically، وفرضت ذلك الحين. يتم توصيل الشرايين والقسطرة الوريدية إلى جهاز القلب الرئة والكبد perfused لمع ملفلان (1 ملغ / كغ من وزن الجسم) لمدة 60 دقيقة. بهذه الطريقة فمن الممكن أن يروي محليا الكبد مع جرعة عالية من العلاج الكيميائي وكيل، دون تسرب إلى الدوران الجهازي. في الدراسات السابقة بما في ذلك المرضى الذين يعانون من الانبثاث الكبد معزولة من سرطان الجلد العنبية، وهو معدل الاستجابة الكلي من 33-100٪ ومتوسط ​​بقاء ما بين 9 و 13 شهرا،تم الإبلاغ. والهدف من هذا البروتوكول هو إعطاء وصف واضح لكيفية تنفيذ الإجراء ومناقشة IHP كخيار لعلاج الكبد الانبثاث من سرطان الجلد العنبي.

وتضمن تقرير أولي 22 مريضا عولجوا ملفلان مع أو بدون TNF-ألفا. أظهرت النتائج معدل الاستجابة الكلي 62٪، من بينهم اثنان من المرضى الذين يعانون من CR (10٪)، ومتوسط ​​البقاء الإجمالي 11 شهرا 20. وأفادت دراسة لاحقة على نتائج 29 مريضا على التوالي تعامل باستخدام ملفلان وحدها. وك?...

Watch this Scientific Journal Video about Isolated Hepatic Perfusion as a Treatment for Liver Metastases of Uveal Melanoma at JoVE.com

Isolated Hepatic Perfusion as a Treatment 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.

معزولة كبدي الإرواء كعلاج للالانبثاث الكبد من العنبية الميلانوما

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

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13.3K Views.  Institute of Clinical Sciences. 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.

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

Murine Model of Metastatic Liver Tumors in the Setting of Ischemia Reperfusion Injury

Liver ischemia and reperfusion (I/R) injury, a common clinical challenge, remains an inevitable pathophysiological process that has been shown to induce multiple tissue and organ damage. Despite recent advances and therapeutic approaches, the overall morbidity has remained unsatisfactory especially in patients with underlying parenchymal abnormalities. In the context of aggressive cancer growth and metastasis, surgical I/R is suspected to be the promoter regulating tumor recurrence. This article aims to describe a clinically relevant murine model of liver I/R and colorectal liver metastasis. In doing so, we aim to assist other investigators in establishing and perfecting this model for their routine research practice to better understand the effects of liver I/R on promoting liver metastases.

We describe in detail a clinically relevant colorectal cancer liver metastases (CRLM) tumor model and the influence of liver ischemia reperfusion (I/R) in tumor growth and metastasis. This model can help to better understand the mechanisms underlying surgery-induced promotion of liver metastatic growth.

نموذج مورين من أورام الكبد النقيلي في وضع إصابة نقص تروية التسريب

Liver ischemia and reperfusion (I/R) injury, a common clinical challenge, remains an inevitable pathophysiological process that has been shown to induce ...

Cancer Research

Generation of a Liver Orthotopic Human Uveal Melanoma Xenograft Platform in Immunodeficient Mice

In recent decades, subcutaneously implanted patient-derived xenograft tumors or cultured human cell lines have been increasingly recognized as more representative models to study human cancers in immunodeficient mice than traditional established human cell lines in vitro. Recently, orthotopically implanted patient-derived tumor xenograft (PDX) models in mice have been developed to better replicate features of patient tumors. A liver orthotopic xenograft mouse model is expected to be a useful cancer research platform, providing insights into tumor biology and drug therapy. However, liver orthotopic tumor implantation is generally complicated. Here we describe our protocols for the orthotopic implantation of patient-derived liver-metastatic uveal melanoma tumors. We cultured human liver metastatic uveal melanoma cell lines into immunodeficient mice. The protocols can result in consistently high technical success rates using either a surgical orthotopic implantation technique with chunks of patient-derived uveal melanoma tumor or a needle injection technique with cultured human cell line. We also describe protocols for CT scanning to detect interior liver tumors and for re-implantation techniques using cryopreserved tumors to achieve re-engraftment. Together, these protocols provide a better platform for liver orthotopic tumor mouse models of liver metastatic uveal melanoma in translational research.

Orthotopic human liver metastatic uveal melanoma xenograft mouse models were created using surgical orthotopic implantation techniques with patient-derived tumor chunk and needle injection techniques with cultured human uveal melanoma cell lines.

جيل من الكبد البشرية العظام الإنسان الجلد البقري النظام الأساسي في الفئران نقص المناعة

In recent decades, subcutaneously implanted patient-derived xenograft tumors or cultured human cell lines have been increasingly recognized as more ...

Portal Vein Injection of Colorectal Cancer Organoids to Study the Liver Metastasis Stroma

Hepatic metastasis of colorectal cancer (CRC) is a leading cause of cancer-related death. Cancer-associated fibroblasts (CAFs), a major component of the tumor microenvironment, play a crucial role in metastatic CRC progression and predict poor patient prognosis. However, there is a lack of satisfactory mouse models to study the crosstalk between metastatic cancer cells and CAFs. Here, we present a method to investigate how liver metastasis progression is regulated by the metastatic niche and possibly could be restrained by stroma-directed therapy. Portal vein injection of CRC organoids generated a desmoplastic reaction, which faithfully recapitulated the fibroblast-rich histology of human CRC liver metastases. This model was tissue-specific with a higher tumor burden in the liver when compared to an intra-splenic injection model, simplifying mouse survival analyses. By injecting luciferase-expressing tumor organoids, tumor growth kinetics could be monitored by in vivo imaging. Moreover, this preclinical model provides a useful platform to assess the efficacy of therapeutics targeting the tumor mesenchyme. We describe methods to examine whether adeno-associated virus-mediated delivery of a tumor-inhibiting stromal gene to hepatocytes could remodel the tumor microenvironment and improve mouse survival. This approach enables the development and assessment of novel therapeutic strategies to inhibit hepatic metastasis of CRC.

Portal vein injection of colorectal cancer (CRC) organoids generates stroma-rich liver metastasis. This mouse model of CRC hepatic metastasis represents a useful tool to study tumor-stroma interactions and develop novel stroma-directed therapeutics such as adeno-associated virus-mediated gene therapies.

حقن الوريد البابي لسرطان القولون والمستقيم العضوي لدراسة سدى ورم خبيث في الكبد

Hepatic metastasis of colorectal cancer (CRC) is a leading cause of cancer-related death. Cancer-associated fibroblasts (CAFs), a major component of the ...

Technique of Subnormothermic Ex Vivo Liver Perfusion for the Storage, Assessment, and Repair of Marginal Liver Grafts

The success of liver transplantation has resulted in a dramatic organ shortage. In most transplant regions 20-30% of patients on the waiting list for liver transplantation die without receiving an organ transplant or are delisted for disease progression. One strategy to increase the donor pool is the utilization of marginal grafts, such as fatty livers, grafts from older donors, or donation after cardiac death (DCD). The current preservation technique of cold static storage is only poorly tolerated by marginal livers resulting in significant organ damage. In addition, cold static organ storage does not allow graft assessment or repair prior to transplantation.
These shortcomings of cold static preservation have triggered an interest in warm perfused organ preservation to reduce cold ischemic injury, assess liver grafts during preservation, and explore the opportunity to repair marginal livers prior to transplantation. The optimal pressure and flow conditions, perfusion temperature, composition of the perfusion solution and the need for an oxygen carrier has been controversial in the past.
In spite of promising results in several animal studies, the complexity and the costs have prevented a broader clinical application so far. Recently, with enhanced technology and a better understanding of liver physiology during ex vivo perfusion the outcome of warm liver perfusion has improved and consistently good results can be achieved.
This paper will provide information about liver retrieval, storage techniques, and isolated liver perfusion in pigs. We will illustrate a) the requirements to ensure sufficient oxygen supply to the organ, b) technical considerations about the perfusion machine and the perfusion solution, and c) biochemical aspects of isolated organs.

Technique of Subnormothermic Ex Vivo Liver Perfusion for the Storage, Assessment, and Repair of Marginal Liver Grafts

Marginal grafts, such as fatty livers, grafts from older donors, or livers retrieved after cardiac death (DCD) tolerate conventional, cold static storage only poorly. We developed a novel model of subnormothermic ex vivo liver perfusion for preservation, assessment, and repair of marginal liver grafts prior to transplantation.

تقنية Subnormothermic<em&gt; فيفو السابقين</em&gt; الكبد الإرواء للتخزين، تقييم، وإصلاح الطعوم الكبد الهامشية

The success of liver transplantation has resulted in a dramatic organ shortage. In most transplant regions 20-30% of patients on the waiting list for ...

Implantation and Evaluation of Melanoma in the Murine Choroid via Optical Coherence Tomography

Establishing experimental choroidal melanoma models is challenging in terms of the ability to induce tumors at the correct localization. In addition, difficulties in observing posterior choroidal melanoma in vivo limit tumor location and growth evaluation in real-time. The approach described here optimizes techniques for establishing choroidal melanoma in mice via a multi-step sub-choroidal B16LS9 cell injection procedure. To enable precision in injecting into the small dimensions of the mouse uvea, the complete procedure is performed under a microscope. First, a conjunctival peritomy is formed in the dorsal-temporal area of the eye. Then, a tract into the sub-choroidal space is created by inserting a needle through the exposed sclera. This is followed by the insertion of a blunt needle into the tract and the injection of melanoma cells into the choroid. Immediately after injection, noninvasive optical coherence tomography (OCT) imaging is utilized to determine tumor location and progress. Retinal detachment is evaluated as a predictor of tumor site and size. The presented method enables the reproducible induction of choroid-localized melanoma in mice and the live imaging of tumor growth evaluation. As such, it provides a valuable tool for studying intraocular tumors.

Implantation and Evaluation of Melanoma in the Murine Choroid via Optical Coherence Tomography

The present protocol describes the implantation and evaluation of melanoma in the murine choroid utilizing optical coherence tomography.

زرع وتقييم سرطان الجلد في المشيمية الفئران عن طريق التصوير المقطعي للتماسك البصري

Establishing experimental choroidal melanoma models is challenging in terms of the ability to induce tumors at the correct localization. In addition, ...

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.

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.

عن طريق الجلد كبدي الإرواء (PHP) مع ملفلان كعلاج للالغير قطوعة الانبثاث تقتصر على الكبد

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

Visualization of Vascular and Parenchymal Regeneration after 70% Partial Hepatectomy in Normal Mice

A modified silicone injection procedure was used for visualization of the hepatic vascular tree. This procedure consisted of in-vivo injection of the silicone compound, via a 26 G catheter, into the portal or hepatic vein. After silicone injection, organs were explanted and prepared for ex-vivo micro-CT (&#181;CT) scanning. The silicone injection procedure is technically challenging. Achieving a successful outcome requires extensive microsurgical experience from the surgeon. One of the challenges of this procedure involves determining the adequate perfusion rate for the silicone compound. The perfusion rate for the silicone compound needs to be defined based on the hemodynamic of the vascular system of interest. Inappropriate perfusion rate can lead to an incomplete perfusion, artificial dilation and rupturing of vascular trees.
The 3D reconstruction of the vascular system was based on CT scans and was achieved using preclinical software such as HepaVision. The quality of the reconstructed vascular tree was directly related to the quality of silicone perfusion. Subsequently computed vascular parameters indicative of vascular growth, such as total vascular volume, were calculated based on the vascular reconstructions. Contrasting the vascular tree with silicone allowed for subsequent histological work-up of the specimen after &#181;CT scanning. The specimen can be subjected to serial sectioning, histological analysis and whole slide scanning, and thereafter to 3D reconstruction of the vascular trees based on histological images. This is the prerequisite for the detection of molecular events and their distribution with respect to the vascular tree. This modified silicone injection procedure can also be used to visualize and reconstruct the vascular systems of other organs. This technique has the potential to be extensively applied to studies concerning vascular anatomy and growth in various animal and disease models.

Tools used for visualizing vascular regeneration require methods for contrasting the vascular trees. This film demonstrated a delicate injection technique used to achieve optimal contrasting of the vascular trees and illustrate the potential benefits resulting from a detailed analysis of the resulting specimen using &#181;CT and histological serial sections.

التصور الأوعية الدموية ومتني التجديد بعد استئصال الكبد الجزئي 70٪ في الفئران العادية

A modified silicone injection procedure was used for visualization of the hepatic vascular tree. This procedure consisted of in-vivo injection of the ...

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.

نموذج حيوانات صغيرة من نضح الكبد نورموثيرميك السابقين فيفو

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

Isolation of Regenerating Hepatocytes after Partial Hepatectomy in Mice

Partial hepatectomy has been widely used to investigate liver regeneration in mice, but the isolation of high yields of viable hepatocytes for downstream single-cell applications is challenging. A marked accumulation of lipids within regenerating hepatocytes is observed during the first 2 days of normal liver regeneration in mice. This so-called transient regeneration-associated steatosis (TRAS) is temporary but partially overlaps the major proliferative phase. Density-gradient purification is the backbone of most existing protocols for the isolation of primary hepatocytes. As gradient purification relies on the density and size of cells, it separates non-steatotic from steatotic hepatocyte populations. Therefore, fatty hepatocytes often are lost, yielding non-representative hepatocyte fractions. 
The presented protocol describes an easy and reliable method for the in vivo isolation of regenerating hepatocytes regardless of their lipid content. Hepatocytes from male C57BL/6 mice are isolated 24-48 h after hepatectomy by a classic two-step collagenase perfusion approach. A standard peristaltic pump drives the warmed solutions via the catheterized inferior vena cava into the remnant, using a retrograde perfusion technique with outflow through the portal vein. Hepatocytes are dissociated by collagenase for their release from the Glisson's capsule. After washing and careful centrifugation, the hepatocytes can be used for any downstream analyses. In conclusion, this paper describes a straightforward and reproducible technique for the isolation of a representative population of regenerating hepatocytes after partial hepatectomy in mice. The method may also aid the study of fatty liver disease.

Lipid-laden hepatocytes are inherent to liver regeneration but are usually lost upon density-gradient centrifugation. Here, we present an optimized cell isolation protocol that retains steatotic hepatocytes, yielding representative populations of regenerating hepatocytes after partial hepatectomy in mice.

عزل خلايا الكبد المتجددة بعد استئصال الكبد الجزئي في الفئران

Partial hepatectomy has been widely used to investigate liver regeneration in mice, but the isolation of high yields of viable hepatocytes for downstream ...

Development and Validation of a Porcine Ex Situ Isolated Liver Perfusion Model 

Porcine Normothermic Isolated Liver Perfusion

Porcine models of liver ex situ normothermic machine perfusion (NMP) are increasingly being used in transplant research. Contrary to rodents, porcine livers are anatomically and physiologically close to humans, with similar organ size and bile composition. NMP preserves the liver graft at near-to-physiological conditions by recirculating a warm, oxygenated, and nutrient-enriched red blood cell-based perfusate through the liver vasculature. NMP can be used to study ischemia-reperfusion injury, preserve a liver ex situ before transplantation, assess the liver's function prior to implantation, and provide a platform for organ repair and regeneration. Alternatively, NMP with a whole blood-based perfusate can be used to mimic transplantation. Nevertheless, this model is labor-intensive, technically challenging, and carries a high financial cost.
In this porcine NMP model, we use warm ischemic damaged livers (corresponding to donation after circulatory death). First, general anesthesia with mechanical ventilation is initiated, followed by the induction of warm ischemia by clamping the thoracic aorta for 60 min. Cannulas inserted in the abdominal aorta and portal vein allow flush-out of the liver with cold preservation solution. The flushed-out blood is washed with a cell saver to obtain concentrated red blood cells. Following hepatectomy, cannulas are inserted in the portal vein, hepatic artery, and infra-hepatic vena cava and connected to a closed perfusion circuit primed with a plasma expander and red blood cells. A hollow fiber oxygenator is included in the circuit and coupled to a heat exchanger to maintain a pO2 of 70-100 mmHg at 38 &#176;C. NMP is achieved by a continuous flow directly through the artery and via a venous reservoir through the portal vein. Flows, pressures, and blood gas values are continuously monitored. To evaluate the liver injury, perfusate and tissue are sampled at predefined time points; bile is collected via a cannula in the common bile duct.

The porcine model of liver normothermic machine perfusion (NMP), described here, can be successfully used to study NMP as a preservation strategy, a tool for viability assessment, and a platform for organ repair. It holds a high translational value, however it is technically challenging and labor-intensive.

Porcine models of liver ex situ normothermic machine perfusion (NMP) are increasingly being used in transplant research. Contrary to rodents, porcine ...