Figure 2.

Experimental design. Four hindlimbs were included in each of the three groups undergoing 48 h preservation and recovery. The experimental group underwent supercooling at – 5 °C followed by (1) 2 h SubNormothermic Machine Perfusion (SNMP) recovery and (2) 2 h Normothermic Machine Perfusion (NMP). The control groups 1 and 2 underwent 48 h static cold storage (4 °C, in HTK). Cold Storage + SNMP group received 2 h SNMP before 2 h normothermic reperfusion, while Cold Storage group directly received NMP. All groups received the same monitoring of weight gain, perfusion parameters, biochemical measurements, and histology at each step. HTK: Histidine-Tryptophan-Ketoglutarate solution.