Influence of perfusate on liver viability during hypothermic machine perfusion.
OBJECTIVE
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).


CONCLUSIONS
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.
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
JJ
Jia
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.
Jing
J
Zhang
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.
Jian-Hui
JH
Li
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.
Xu-Dong
XD
Chen
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.
Li
L
Jiang
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.
Yan-Fei
YF
Zhou
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.
Ning
N
He
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.
Hai-Yang
HY
Xie
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.
Lin
L
Zhou
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.
Shu-Sen
SS
Zheng
D000241:Adenosine / Q000494:pharmacology; D000255:Adenosine Triphosphate / Q000378:metabolism; D000410:Alanine Transaminase / Q000378:metabolism; D000493:Allopurinol / Q000494:pharmacology; D000818:Animals; D001219:Aspartate Aminotransferases / Q000378:metabolism; D015415:Biomarkers / Q000378:metabolism; D050377:Cold Ischemia; D003080:Cold Temperature; D005947:Glucose / Q000494:pharmacology; D005978:Glutathione / Q000494:pharmacology; D006498:Hepatectomy; D007328:Insulin / Q000494:pharmacology; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008099:Liver / Q000187:drug effects* / Q000201:enzymology / Q000473:pathology; D008111:Liver Function Tests; D008297:Male; D008315:Malondialdehyde / Q000378:metabolism; D008353:Mannitol / Q000494:pharmacology; D009926:Organ Preservation / Q000379:methods*; D019188:Organ Preservation Solutions / Q000494:pharmacology*; D010477:Perfusion / Q000379:methods*; D011189:Potassium Chloride / Q000494:pharmacology; D011343:Procaine / Q000494:pharmacology; D011887:Raffinose / Q000494:pharmacology; D017207:Rats, Sprague-Dawley; D013997:Time Factors; D014022:Tissue Survival
D015415:Biomarkers; C039984:Bretschneider cardioplegic solution; D007328:Insulin; D019188:Organ Preservation Solutions; C055206:University of Wisconsin-lactobionate solution; D008353:Mannitol; D008315:Malondialdehyde; D011343:Procaine; D000493:Allopurinol; D011189:Potassium Chloride; D000255:Adenosine Triphosphate; D007770:L-Lactate Dehydrogenase; D001219:Aspartate Aminotransferases; D000410:Alanine Transaminase; D005978:Glutathione; D005947:Glucose; D000241:Adenosine; D011887:Raffinose
Histidine-tryptophan-ketoglutarate solution; Hypothermic machine perfusion; Liver viability; Static cold storage; Wisconsin cold-storage solution