Scanning Electron Microscopy Findings of Machine Perfused Liver Graft After Warm Ischemia Between Hypothermic and Rewarming Machine Perfusion in Pigs
The shortage of organ donors is a universal problem. Use of grafts from donors after cardiac death would greatly contribute to the expansion of the donor organ pool. The two major methods of preservation are cold storage and machine perfusion (MP) preservation, and each has its own advantages. Sever...
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Published in | Transplantation proceedings Vol. 48; no. 7; pp. 2467 - 2470 |
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Main Authors | , , , , , , , |
Format | Journal Article |
Language | English |
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Elsevier Inc
01.09.2016
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ISSN | 0041-1345 1873-2623 1873-2623 |
DOI | 10.1016/j.transproceed.2016.03.059 |
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Abstract | The shortage of organ donors is a universal problem. Use of grafts from donors after cardiac death would greatly contribute to the expansion of the donor organ pool. The two major methods of preservation are cold storage and machine perfusion (MP) preservation, and each has its own advantages. Several studies have reported the relative merits of MP for the preservation for grafts from donors after cardiac death. In this study, we used scanning electron microscopy (SEM) to assess the damage to the liver between hypothermic and rewarming preservation conditions.
Porcine livers were perfused with a newly developed MP system. The livers were perfused for 4 hours with a modified University of Wisconsin solution–gluconate solution. In group 1, grafts were preserved with warm ischemic time for 60 minutes and hypothermic machine perfusion (HMP) for 4 hours. In group 2, grafts were preserved with warn ischemic time for 60 minutes and had rewarming up to 22°C by MP (RMP) for 4 hours.
A significant enlargement of the mitochondria were observed in both the HMP and RMP groups under higher magnification, Additionally, vacuoles appeared occasionally in hepatocytes in the RMP for 4 hours group, but not in the HMP for 4 hours group.
An analysis by scanning electron microscope appears to be useful to evaluate the levels of damage of hepatocytes compared with transmission electron microscopy, and further study is needed to analyze the significance of the appearance of swelling of mitochondria and vacuolization during preservation.
•The use of grafts from donors after cardiac death would greatly contribute to the expansion of the donor organ pool.•A novel machine perfusion preservation system to control the perfusate temperatures from hypothermic to subnormothermic conditions is presented.•Scanning electron microscopy of tissue samples from a liver biopsy can clearly show the internal structures of intracellular organelles compared to transmission electron microscopy. |
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AbstractList | The shortage of organ donors is a universal problem. Use of grafts from donors after cardiac death would greatly contribute to the expansion of the donor organ pool. The two major methods of preservation are cold storage and machine perfusion (MP) preservation, and each has its own advantages. Several studies have reported the relative merits of MP for the preservation for grafts from donors after cardiac death. In this study, we used scanning electron microscopy (SEM) to assess the damage to the liver between hypothermic and rewarming preservation conditions.
Porcine livers were perfused with a newly developed MP system. The livers were perfused for 4 hours with a modified University of Wisconsin solution-gluconate solution. In group 1, grafts were preserved with warm ischemic time for 60 minutes and hypothermic machine perfusion (HMP) for 4 hours. In group 2, grafts were preserved with warn ischemic time for 60 minutes and had rewarming up to 22°C by MP (RMP) for 4 hours.
A significant enlargement of the mitochondria were observed in both the HMP and RMP groups under higher magnification, Additionally, vacuoles appeared occasionally in hepatocytes in the RMP for 4 hours group, but not in the HMP for 4 hours group.
An analysis by scanning electron microscope appears to be useful to evaluate the levels of damage of hepatocytes compared with transmission electron microscopy, and further study is needed to analyze the significance of the appearance of swelling of mitochondria and vacuolization during preservation. The shortage of organ donors is a universal problem. Use of grafts from donors after cardiac death would greatly contribute to the expansion of the donor organ pool. The two major methods of preservation are cold storage and machine perfusion (MP) preservation, and each has its own advantages. Several studies have reported the relative merits of MP for the preservation for grafts from donors after cardiac death. In this study, we used scanning electron microscopy (SEM) to assess the damage to the liver between hypothermic and rewarming preservation conditions.BACKGROUNDThe shortage of organ donors is a universal problem. Use of grafts from donors after cardiac death would greatly contribute to the expansion of the donor organ pool. The two major methods of preservation are cold storage and machine perfusion (MP) preservation, and each has its own advantages. Several studies have reported the relative merits of MP for the preservation for grafts from donors after cardiac death. In this study, we used scanning electron microscopy (SEM) to assess the damage to the liver between hypothermic and rewarming preservation conditions.Porcine livers were perfused with a newly developed MP system. The livers were perfused for 4 hours with a modified University of Wisconsin solution-gluconate solution. In group 1, grafts were preserved with warm ischemic time for 60 minutes and hypothermic machine perfusion (HMP) for 4 hours. In group 2, grafts were preserved with warn ischemic time for 60 minutes and had rewarming up to 22°C by MP (RMP) for 4 hours.METHODSPorcine livers were perfused with a newly developed MP system. The livers were perfused for 4 hours with a modified University of Wisconsin solution-gluconate solution. In group 1, grafts were preserved with warm ischemic time for 60 minutes and hypothermic machine perfusion (HMP) for 4 hours. In group 2, grafts were preserved with warn ischemic time for 60 minutes and had rewarming up to 22°C by MP (RMP) for 4 hours.A significant enlargement of the mitochondria were observed in both the HMP and RMP groups under higher magnification, Additionally, vacuoles appeared occasionally in hepatocytes in the RMP for 4 hours group, but not in the HMP for 4 hours group.RESULTSA significant enlargement of the mitochondria were observed in both the HMP and RMP groups under higher magnification, Additionally, vacuoles appeared occasionally in hepatocytes in the RMP for 4 hours group, but not in the HMP for 4 hours group.An analysis by scanning electron microscope appears to be useful to evaluate the levels of damage of hepatocytes compared with transmission electron microscopy, and further study is needed to analyze the significance of the appearance of swelling of mitochondria and vacuolization during preservation.CONCLUSIONSAn analysis by scanning electron microscope appears to be useful to evaluate the levels of damage of hepatocytes compared with transmission electron microscopy, and further study is needed to analyze the significance of the appearance of swelling of mitochondria and vacuolization during preservation. The shortage of organ donors is a universal problem. Use of grafts from donors after cardiac death would greatly contribute to the expansion of the donor organ pool. The two major methods of preservation are cold storage and machine perfusion (MP) preservation, and each has its own advantages. Several studies have reported the relative merits of MP for the preservation for grafts from donors after cardiac death. In this study, we used scanning electron microscopy (SEM) to assess the damage to the liver between hypothermic and rewarming preservation conditions. Porcine livers were perfused with a newly developed MP system. The livers were perfused for 4 hours with a modified University of Wisconsin solution–gluconate solution. In group 1, grafts were preserved with warm ischemic time for 60 minutes and hypothermic machine perfusion (HMP) for 4 hours. In group 2, grafts were preserved with warn ischemic time for 60 minutes and had rewarming up to 22°C by MP (RMP) for 4 hours. A significant enlargement of the mitochondria were observed in both the HMP and RMP groups under higher magnification, Additionally, vacuoles appeared occasionally in hepatocytes in the RMP for 4 hours group, but not in the HMP for 4 hours group. An analysis by scanning electron microscope appears to be useful to evaluate the levels of damage of hepatocytes compared with transmission electron microscopy, and further study is needed to analyze the significance of the appearance of swelling of mitochondria and vacuolization during preservation. •The use of grafts from donors after cardiac death would greatly contribute to the expansion of the donor organ pool.•A novel machine perfusion preservation system to control the perfusate temperatures from hypothermic to subnormothermic conditions is presented.•Scanning electron microscopy of tissue samples from a liver biopsy can clearly show the internal structures of intracellular organelles compared to transmission electron microscopy. Abstract Background The shortage of organ donors is a universal problem. Use of grafts from donors after cardiac death would greatly contribute to the expansion of the donor organ pool. The two major methods of preservation are cold storage and machine perfusion (MP) preservation, and each has its own advantages. Several studies have reported the relative merits of MP for the preservation for grafts from donors after cardiac death. In this study, we used scanning electron microscopy (SEM) to assess the damage to the liver between hypothermic and rewarming preservation conditions. Methods Porcine livers were perfused with a newly developed MP system. The livers were perfused for 4 hours with a modified University of Wisconsin solution–gluconate solution. In group 1, grafts were preserved with warm ischemic time for 60 minutes and hypothermic machine perfusion (HMP) for 4 hours. In group 2, grafts were preserved with warn ischemic time for 60 minutes and had rewarming up to 22°C by MP (RMP) for 4 hours. Results A significant enlargement of the mitochondria were observed in both the HMP and RMP groups under higher magnification, Additionally, vacuoles appeared occasionally in hepatocytes in the RMP for 4 hours group, but not in the HMP for 4 hours group. Conclusions An analysis by scanning electron microscope appears to be useful to evaluate the levels of damage of hepatocytes compared with transmission electron microscopy, and further study is needed to analyze the significance of the appearance of swelling of mitochondria and vacuolization during preservation. |
Author | Matsuno, N. Meng, L. Bochimoto, H. Watanabe, T. Obara, H. Furukori, M. Fukukawa, H. Watanabe, K. |
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References_xml | – volume: 84 start-page: 105 year: 2002 end-page: 111 ident: bib5 article-title: Mitochondria: regulating the inevitable publication-title: Biochimie – volume: 133 start-page: 213 year: 1984 end-page: 222 ident: bib4 article-title: A preparation method for observing intracellular structures by scanning electron microscopy publication-title: J Microsc – volume: 45 start-page: 1684 year: 2013 end-page: 1689 ident: bib2 article-title: Impact of rewarming preservation by continuous machine perfusion: improved post-transplant recovery in pigs publication-title: Transplant Proc – volume: 10 start-page: 372 year: 2010 end-page: 381 ident: bib1 article-title: Hypothermic machine preservation in human liver transplantation: the first clinical series publication-title: Am J Transplant – volume: 3 start-page: 527 year: 2003 end-page: 535 ident: bib6 article-title: Role of the mitochondrial permeability transition in apoptotic and necrotic death after ischemia/reperfusion injury to hepatocytes publication-title: Curr Mol Med – volume: 45 start-page: 1684 year: 2013 end-page: 1689 ident: bib3 article-title: Rewarming preservation by organ perfusion system for donation after cardiac death liver grafts in pigs publication-title: Transplant Proc – volume: 10 start-page: 372 issue: 2 year: 2010 ident: 10.1016/j.transproceed.2016.03.059_bib1 article-title: Hypothermic machine preservation in human liver transplantation: the first clinical series publication-title: Am J Transplant doi: 10.1111/j.1600-6143.2009.02932.x – volume: 3 start-page: 527 issue: 6 year: 2003 ident: 10.1016/j.transproceed.2016.03.059_bib6 article-title: Role of the mitochondrial permeability transition in apoptotic and necrotic death after ischemia/reperfusion injury to hepatocytes publication-title: Curr Mol Med doi: 10.2174/1566524033479564 – volume: 45 start-page: 1684 issue: 5 year: 2013 ident: 10.1016/j.transproceed.2016.03.059_bib3 article-title: Rewarming preservation by organ perfusion system for donation after cardiac death liver grafts in pigs publication-title: Transplant Proc – volume: 45 start-page: 1684 year: 2013 ident: 10.1016/j.transproceed.2016.03.059_bib2 article-title: Impact of rewarming preservation by continuous machine perfusion: improved post-transplant recovery in pigs publication-title: Transplant Proc doi: 10.1016/j.transproceed.2013.01.098 – volume: 133 start-page: 213 issue: Pt 2 year: 1984 ident: 10.1016/j.transproceed.2016.03.059_bib4 article-title: A preparation method for observing intracellular structures by scanning electron microscopy publication-title: J Microsc doi: 10.1111/j.1365-2818.1984.tb00487.x – volume: 84 start-page: 105 issue: 2-3 year: 2002 ident: 10.1016/j.transproceed.2016.03.059_bib5 article-title: Mitochondria: regulating the inevitable publication-title: Biochimie doi: 10.1016/S0300-9084(02)01380-9 |
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Snippet | The shortage of organ donors is a universal problem. Use of grafts from donors after cardiac death would greatly contribute to the expansion of the donor organ... Abstract Background The shortage of organ donors is a universal problem. Use of grafts from donors after cardiac death would greatly contribute to the... |
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SubjectTerms | Animals Cryopreservation - methods Death Liver - pathology Liver - ultrastructure Liver Transplantation - methods Microscopy, Electron, Scanning Models, Animal Organ Preservation - methods Organ Preservation Solutions Perfusion - methods Rewarming - methods Surgery Sus scrofa Swine Warm Ischemia - methods |
Title | Scanning Electron Microscopy Findings of Machine Perfused Liver Graft After Warm Ischemia Between Hypothermic and Rewarming Machine Perfusion in Pigs |
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