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©The Author(s) 2015. Published by Baishideng Publishing Group Inc. All rights reserved.
Influence of perfusate on liver viability during hypothermic machine perfusion
Jun-Jun Jia, Jing Zhang, Jian-Hui Li, Xu-Dong Chen, Li Jiang, Yan-Fei Zhou, Ning He, Hai-Yang Xie, Lin Zhou, Shu-Sen Zheng
Jun-Jun Jia, Jing Zhang, Jian-Hui Li, Ning He, Hai-Yang Xie, Lin Zhou, Shu-Sen Zheng, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, The First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou 310003, Zhejiang Province, China
Jun-Jun Jia, Jing Zhang, Jian-Hui Li, Li Jiang, Ning He, Hai-Yang Xie, Lin Zhou, Shu-Sen Zheng, Key Laboratory of Combined Multiorgan Transplantation, Ministry of Public Health, Zhejiang University School of Medicine, Hangzhou 310003, Zhejiang Province, China
Jun-Jun Jia, Jing Zhang, Jian-Hui Li, Li Jiang, Ning He, Hai-Yang Xie, Lin Zhou, Shu-Sen Zheng, Department of Hepatobiliary and Pancreatic Surgery, First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou 310003, Zhejiang Province, China
Xu-Dong Chen, Yan-Fei Zhou, Department of Vascular Surgery, First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou 310003, Zhejiang Province, China
Author contributions: Jia JJ and Zhang J contributed equally to this work; Zhou L, Li JH and Zheng SS conceived and designed the experiments; Chen XD and Xie HY contributed reagents/materials/analytic tools; Jia JJ and Zhang J wrote the manuscript; Jia JJ, Zhang J, Jiang L, Zhou YF and He N performed the experiments; Jia JJ and Zhang J analyzed the data.
Supported by National Science and Technology Major Project, No. 2012ZX10002-017; Natural Science Foundation of China for Innovative Research Group, No. 81121002; National Natural Science Foundation of China, No. 81000137 and No. 81470891; The Qianjiang Talent Program of Zhejiang Province, China, No. 2012R10045; the Scientific Research Program for the Returned Overseas Chinese Scholars, Ministry of Health, China, No. J20112008; National High Technology Research and Development Program of China for Young Scientists (863 Program), No. 2015AA020923; and Ministry of Education, Zhejiang Province, China, No. Y201328095.
Institutional review board statement: The study was reviewed and approved by the Zhejiang University Institutional Review Board.
Institutional animal care and use committee statement: All procedures involving animals were reviewed and approved by the Institutional Animal Care and Use Committee of Zhejiang University.
Conflict-of-interest statement: No benefits in any form have been received or will be received from a commercial party related directly or indirectly to the subject of this article.
Data sharing statement: Technical appendix, statistical code, and dataset available from the corresponding author at shusenzheng@zju.edu.cn or surgeonlee@126.com
Open-Access: This article is an open-access article which was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution Non Commercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See: http://creativecommons.org/licenses/by-nc/4.0/
Correspondence to: Shu-Sen Zheng, MD, PhD, FACS, Department of Hepatobiliary and Pancreatic Surgery, First Affiliated Hospital, Zhejiang University School of Medicine, No. 79 Qingchun Road, Hangzhou 310003, Zhejiang Province, China. shusenzheng@zju.edu.cn
Telephone: +86-571-87236567 Fax: +86-571-87236884
Received: February 11, 2015
Peer-review started: February 12, 2015
First decision: March 26, 2015
Revised: April 12, 2015
Accepted: June 9, 2015
Article in press: June 10, 2015
Published online: August 7, 2015
AIM: To optimize the perfusates used for hypothermic machine perfusion (HMP).
METHODS: Sprague-Dawley rats were assigned randomly to three groups (n = 12 per group) that received either saline, University of Wisconsin cold-storage solution (UW) or histidine-tryptophan-ketoglutarate solution (HTK) as the perfusate. Each group was divided into two subgroups: static cold storage (SCS) and HMP (n = 6 per subgroup). The liver graft was retrieved according to the method described by Kamada. For the SCS group, the graft was directly placed into cold perfusate (0-4 °C) for 6 h after liver isolation while the portal vein of the graft was connected to the perfusion machine for the HMP group. Then the perfusates were collected at different time points for analysis of aspartate aminotransferase (AST), alanine transaminase (ALT) and lactate dehydrogenase (LDH) levels. Liver tissues were obtained for evaluation of histology, dry/wet weight (D/W) ratio, and malondialdehyde (MDA) and adenosine-triphosphate (ATP) levels. The portal vein pressure and velocity were monitored in real time in all HMP subgroups.
RESULTS: Comparison of HMP and SCS: Regardless of the perfusate, HMP improved the architecture of donor graft in reducing the congestion around sinusoids and central vein and maintaining sinusoid lining in morphology; HMP improved liver function in terms of ALT, AST and LDH, especially during the 3-6 h period (SCS vs HMP using saline: ALT3, 225.00 ± 105.62 vs 49.50 ± 18.50, P = 0.047; LDH3, 1362.17 ± 563.30 vs 325.75 ± 147.43, P = 0.041; UW: LDH6, 2880.14 ± 948.46 vs 2135.00 ± 174.27, P = 0.049; HTK, AST6, 307.50 ± 52.95 vs 185.20 ± 20.46, P = 0.041); HMP decreased MDA level (saline, 2.79 ± 0.30 vs 1.09 ± 0.09, P = 0.008; UW, 3.01 ± 0.77 vs 1.23 ± 0.68, P = 0.005; HTK, 3.30 ± 0.52 vs 1.56 ± 0.22, P = 0.006). Comparison among HMP subgroups: HTK showed less portal vein resistance than UW and saline (vs saline, 3.41 ± 0.49 vs 5.00 ± 0.38, P < 0.001; vs UW, 3.41 ± 0.49 vs 4.52 ± 0.63, P = 0.007); UW reduced edema most efficiently (vs saline, 0.68 ± 0.02 vs 0.79 ± 0.05, P = 0.013), while HTK maintained ATP levels best (vs saline, 622.60 ± 29.11 vs 327.43 ± 44.66, P < 0.001; vs UW, 622.60 ± 29.11 vs 301.80 ± 37.68, P < 0.001).
CONCLUSION: HMP is superior to SCS in maintaining both architecture and function of liver grafts. Further, HTK was found to be the optimal perfusate for HMP.
Core tip: Although static cold storage (SCS) is the gold standard for liver transplantation, hypothermic machine perfusion (HMP) is currently challenging the limitations of SCS. However, there is no consensus on the basic setting for HMP, including the ideal perfusate. Here we compared the most common preservation solutions [University of Wisconsin cold-storage solution and histidine-tryptophan-ketoglutarate solution (HTK), saline as control] and found that HMP is superior to SCS regardless of different solutions and HTK seems to be the optimal perfusate for HMP.
