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Oil Red O-assessed macrosteatosis in liver transplant donor biopsies predicts ischemia–reperfusion injury and clinical outcome

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Abstract

Steatosis in donor livers is an accepted adverse prognostic factor after liver transplantation. While its semiquantitative assessment shows varying reproducibility, it is questioned as a standard method. Additionally, the influence of hepatic steatosis on ischemia/reperfusion injury (I/R injury) has not been evaluated in biopsies after reperfusion. We compared different staining and analyzing methods for the assessment of donor liver steatosis in order to predict I/R injury and clinical outcome after transplantation. To do this, 56 paired pre- and post-reperfusion liver biopsies were analyzed for macro- (MaS)/micro- (MiS) and total steatosis in cryo and permanent sections by special fat (Oil Red O or ORO) and standard stains. Computerized morphometrical analyses were compared to the semiquantitative assessment by a pathologist. I/R injury was determined histopathologically and by M30 immunohistochemistry. We found ORO to be more sensitive in detecting hepatic steatosis with higher reproducibility for MaS. Semiquantitative analyses were highly reproducible and not inferior to computerized morphometry. Categorized MaS as determined by ORO correlated with the extent of I/R injury, initial poor function, liver enzymes, and survival. Therefore fat stains like ORO are a reliable and easy method comprising significant advantages in the evaluation of hepatic steatosis and are thereby of prognostic value. Computerized analysis is a precise tool though not superior to semiquantitative analyses.

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References

  1. Soejima Y, Shimada M, Suehiro T, Kishikawa K, Yoshizumi T, Hashimoto K, Minagawa R, Hiroshige S, Terashi T, Ninomiya M, Shiotani S, Harada N, Sugimachi K (2003) Use of steatotic graft in living-donor liver transplantation. Transplantation 76(2):344–348. doi:10.1097/01.TP.0000071205.52835.A4

    Article  PubMed  Google Scholar 

  2. Sotiropoulos GC, Paul A, Molmenti E, Lang H, Frilling A, Napieralski BP, Nadalin S, Treckmann J, Brokalaki EI, Gerling T, Broelsch CE, Malago M (2005) Liver transplantation for hepatocellular carcinoma in cirrhosis within the Eurotransplant area: an additional option with "livers that nobody wants". Transplantation 80(7):897–902

    Article  PubMed  Google Scholar 

  3. Rey JW, Wirges U, Dienes HP, Fries JW (2009) Hepatic steatosis in organ donors: disparity between surgery and histology? Transplant Proc 41(6):2557–2560. doi:10.1016/j.transproceed.2009.06.121

    Article  PubMed  CAS  Google Scholar 

  4. Sotiropoulos GC, Paul A, Gerling T, Molmenti EP, Nadalin S, Napieralski BP, Treckmann J, Lang H, Saner F, Frilling A, Broelsch CE, Malago M (2006) Liver transplantation with "rescue organ offers" within the Eurotransplant area: a 2-year report from the University Hospital Essen. Transplantation 82(3):304–309. doi:10.1097/01.tp.0000229447.37333.ed

    Article  PubMed  Google Scholar 

  5. Clavien PA, Petrowsky H, DeOliveira ML, Graf R (2007) Strategies for safer liver surgery and partial liver transplantation. N Engl J Med 356(15):1545–1559. doi:10.1056/NEJMra065156

    Article  PubMed  Google Scholar 

  6. Fiorentino M, Vasuri F, Ravaioli M, Ridolfi L, Grigioni WF, Pinna AD, D'Errico-Grigioni A (2009) Predictive value of frozen-section analysis in the histological assessment of steatosis before liver transplantation. Liver Transpl 15(12):1821–1825. doi:10.1002/lt.21948

    Article  PubMed  Google Scholar 

  7. Lo IJ, Lefkowitch JH, Feirt N, Alkofer B, Kin C, Samstein B, Guarrera JV, Renz JF (2008) Utility of liver allograft biopsy obtained at procurement. Liver Transpl 14(5):639–646. doi:10.1002/lt.21419

    Article  PubMed  Google Scholar 

  8. Selzner M, Clavien PA (2001) Fatty liver in liver transplantation and surgery. Semin Liver Dis 21(1):105–113. doi:10.1055/s-2001-12933

    Article  PubMed  CAS  Google Scholar 

  9. Yoong KF, Gunson BK, Neil DA, Mirza DF, Mayer AD, Buckels JA, McMaster P (1999) Impact of donor liver microvesicular steatosis on the outcome of liver retransplantation. Transplant Proc 31(1–2):550–551

    Article  PubMed  CAS  Google Scholar 

  10. Fishbein TM, Fiel MI, Emre S, Cubukcu O, Guy SR, Schwartz ME, Miller CM, Sheiner PA (1997) Use of livers with microvesicular fat safely expands the donor pool. Transplantation 64(2):248–251

    Article  PubMed  CAS  Google Scholar 

  11. Urena MA, Ruiz-Delgado FC, Gonzalez EM, Segurola CL, Romero CJ, Garcia IG, Gonzalez-Pinto I, Gomez Sanz R (1998) Assessing risk of the use of livers with macro and microsteatosis in a liver transplant program. Transplant Proc 30(7):3288–3291

    Article  PubMed  CAS  Google Scholar 

  12. Frongillo F, Avolio AW, Nure E, Mule A, Pepe G, Magalini SC, Agnes S (2009) Quantification of degree of steatosis in extended criteria donor grafts with standardized histologic techniques: implications for graft survival. Transplant Proc 41(4):1268–1272. doi:10.1016/j.transproceed.2009.03.096

    Article  PubMed  CAS  Google Scholar 

  13. El-Badry AM, Breitenstein S, Jochum W, Washington K, Paradis V, Rubbia-Brandt L, Puhan MA, Slankamenac K, Graf R, Clavien PA (2009) Assessment of hepatic steatosis by expert pathologists: the end of a gold standard. Ann Surg 250(5):691–697. doi:10.1097/SLA.0b013e3181bcd6dd

    Article  PubMed  Google Scholar 

  14. Nanashima A, Pillay P, Verran DJ, Painter D, Nakasuji M, Crawford M, Shi L, Ross AG (2002) Analysis of initial poor graft function after orthotopic liver transplantation: experience of an Australian single liver transplantation center. Transplant Proc 34(4):1231–1235

    Article  PubMed  CAS  Google Scholar 

  15. Ploeg RJ, D'Alessandro AM, Knechtle SJ, Stegall MD, Pirsch JD, Hoffmann RM, Sasaki T, Sollinger HW, Belzer FO, Kalayoglu M (1993) Risk factors for primary dysfunction after liver transplantation—a multivariate analysis. Transplantation 55(4):807–813

    Article  PubMed  CAS  Google Scholar 

  16. Weigand K, Brost S, Steinebrunner N, Buchler M, Schemmer P, Muller M (2012) Ischemia/reperfusion injury in liver surgery and transplantation: pathophysiology. HPB Surg 2012:176723. doi:10.1155/2012/176723

    Article  PubMed Central  PubMed  Google Scholar 

  17. Longerich T, Schirmacher P (2006) General aspects and pitfalls in liver transplant pathology. Clin Transplant 20(Suppl 17):60–68. doi:10.1111/j.1399-0012.2006.00602.x

    Article  PubMed  Google Scholar 

  18. Leers MP, Kolgen W, Bjorklund V, Bergman T, Tribbick G, Persson B, Bjorklund P, Ramaekers FC, Bjorklund B, Nap M, Jornvall H, Schutte B (1999) Immunocytochemical detection and mapping of a cytokeratin 18 neo-epitope exposed during early apoptosis. J Pathol 187(5):567–572. doi:10.1002/(SICI)1096-9896(199904)187:5 < 567::AID-PATH288 > 3.0.CO;2-J

    Article  PubMed  CAS  Google Scholar 

  19. McCormack L, Dutkowski P, El-Badry AM, Clavien PA (2011) Liver transplantation using fatty livers: always feasible? J Hepatol 54(5):1055–1062. doi:10.1016/j.jhep.2010.11.004

    Article  PubMed  Google Scholar 

  20. Silva MA (2009) Putting objectivity into assessment of steatosis. Transplantation 88(5):620–621. doi:10.1097/TP.0b013e3181b39085

    Article  PubMed  Google Scholar 

  21. Markin RS, Wisecarver JL, Radio SJ, Stratta RJ, Langnas AN, Hirst K, Shaw BW Jr (1993) Frozen section evaluation of donor livers before transplantation. Transplantation 56(6):1403–1409

    Article  PubMed  CAS  Google Scholar 

  22. McCormack L, Petrowsky H, Jochum W, Mullhaupt B, Weber M, Clavien PA (2007) Use of severely steatotic grafts in liver transplantation: a matched case–control study. Ann Surg 246(6):940–946. doi:10.1097/SLA.0b013e31815c2a3f, discussion 946–948

    Article  PubMed  Google Scholar 

  23. Straub BK, Gyoengyoesi B, Koenig M, Hashani M, Pawella LM, Herpel E, Mueller W, Macher-Goeppinger S, Heid H, Schirmacher P (2013) Adipophilin/perilipin-2 as a lipid droplet-specific marker for metabolically active cells and diseases associated with metabolic dysregulation. Histopathology 62(4):617–631. doi:10.1111/his.12038

    Article  PubMed  Google Scholar 

  24. Imber CJ, St Peter SD, Lopez I, Guiver L, Friend PJ (2002) Current practice regarding the use of fatty livers: a trans-Atlantic survey. Liver Transpl 8(6):545–549. doi:10.1053/jlts.2002.31747

    Article  PubMed  Google Scholar 

  25. Iwasaki M, Takada Y, Hayashi M, Minamiguchi S, Haga H, Maetani Y, Fujii K, Kiuchi T, Tanaka K (2004) Noninvasive evaluation of graft steatosis in living donor liver transplantation. Transplantation 78(10):1501–1505

    Article  PubMed  Google Scholar 

  26. Heuer M, Kaiser GM, Kahraman A, Banysch M, Saner FH, Mathe Z, Gerken G, Paul A, Canbay A, Treckmann JW (2012) Liver transplantation in nonalcoholic steatohepatitis is associated with high mortality and post-transplant complications: a single-center experience. Digestion 86(2):107–113. doi:10.1159/000339344

    Article  PubMed  Google Scholar 

  27. Patkowski W, Zieniewicz K, Skalski M, Krawczyk M (2009) Correlation between selected prognostic factors and postoperative course in liver transplant recipients. Transplant Proc 41(8):3091–3102. doi:10.1016/j.transproceed.2009.09.038

    Article  PubMed  CAS  Google Scholar 

  28. Vetelainen R, van Vliet A, Gouma DJ, van Gulik TM (2007) Steatosis as a risk factor in liver surgery. Ann Surg 245(1):20–30. doi:10.1097/01.sla.0000225113.88433.cf

    Article  PubMed Central  PubMed  Google Scholar 

  29. Cieslak B, Lewandowski Z, Urban M, Ziarkiewicz-Wroblewska B, Krawczyk M (2009) Microvesicular liver graft steatosis as a risk factor of initial poor function in relation to suboptimal donor parameters. Transplant Proc 41(8):2985–2988. doi:10.1016/j.transproceed.2009.08.019

    Article  PubMed  CAS  Google Scholar 

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Acknowledgments

We thank Dorothe Möllmann, Laura Malkus, and Nicole Macha for their skillful technical assistance, and Achim Konietzko for his assistance.

Conflict of interest

The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Institute of Pathology and Neuropathology, University Hospital of Essen, University of Duisburg–Essen, Hufelandstrasse 55, 45122, Essen, Germany

    Henning Reis, Patricia T. Peterek, Jeremias Wohlschlaeger & Hideo Andreas Baba

  2. Department of General, Visceral and Transplantation Surgery, University Hospital of Essen, University of Duisburg–Essen, Hufelandstrasse 55, 45122, Essen, Germany

    Gernot M. Kaiser, Zoltan Mathe, Benjamin Juntermanns, Georgios C. Sotiropoulos, Juergen-Walter Treckmann & Andreas Paul

  3. Carl Zeiss Microscopy GmbH, Königsallee 9-21, 37081, Göttingen, Germany

    Ulrich Beckhove

  4. Department of Gastroenterology and Hepatology, University Hospital of Essen, University of Duisburg–Essen, Hufelandstrasse 55, 45122, Essen, Germany

    Ali Canbay

  5. German Organ Transplantation Foundation (DSO), Lindenallee 29-41, 45127, Essen, Germany

    Ulrike Wirges

  6. Institute for Medical Informatics, Biometry and Epidemiology, University Hospital of Essen, University of Duisburg–Essen, Hufelandstrasse 55, 45122, Essen, Germany

    Andre Scherag

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  1. Henning Reis
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  2. Patricia T. Peterek
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  7. Georgios C. Sotiropoulos
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  13. Andreas Paul
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Correspondence to Hideo Andreas Baba.

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Reis, H., Peterek, P.T., Wohlschlaeger, J. et al. Oil Red O-assessed macrosteatosis in liver transplant donor biopsies predicts ischemia–reperfusion injury and clinical outcome. Virchows Arch 464, 165–174 (2014). https://doi.org/10.1007/s00428-013-1512-3

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  • DOI: https://doi.org/10.1007/s00428-013-1512-3

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