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. 2024 Jun 1;14:12618. doi: 10.1038/s41598-024-63041-8

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© The Author(s) 2024

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Macroscopic and Microscopic results on the porcine VCAs following 48 h preservation and recovery. (a,d,g) The macroscopic aspect of the grafts following 2 h of 37 °C NMP with whole blood was similar in the Cold Storage + SNMP, Cold Storage and Supercooling groups, respectively. The skin paddle presented an ecchymotic aspect, which seemed milder in the Supercooling group, but no significant differences were found histologically. (b,c) At the microscopic level, the Cold storage + SNMP group showed significant interstitial edema, ischemic myocytes (black asterisks) and myocyte fiber injuries (yellow asterisks). (e,f) The Cold storage group showed major edema and mild fiber architecture disruption. (h,i) The Supercooling group showed mild interstitial edema and minor muscle fiber injuries when compared with both control groups. (j) A blinded microscopic score (Kruit et al. 2021) was performed. The overall scores were compared at several time points for each group: T0 (initial), EOP (end of preservation), EOS (end of Steen + SNMP recovery) and EOB (end of NMP blood recovery). A significantly lower muscle injury score was found between the Supercooling group and the Cold storage + SNMP group at the end of the Steen + recovery. The difference between experimental and control groups was not statistically significant at the other time points. (b,e,h) light microscopy (LM), X100, Hematoxylin and Eosin (H&E) staining; (c,f,i) LM X200, H&E.