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Optimization of Ex Vivo Machine Perfusion and Transplantation of Vascularized Composite Allografts
Laura C Burlage1, Alexandre G Lellouch2, Corentin B Taveau3
1Center for Engineering in Medicine and Surgery, Department of Surgery, Massachusetts General Hospital/ Harvard Medical School, Boston, Massachusetts; Department of Surgery, University Medical Center Groningen, Groningen, Netherlands; Vascularized Composite Allotransplantation Laboratory, Center for Transplantation Sciences, Massachusetts General Hospital, Boston, Massachusetts; Division of Plastic and Reconstructive Surgery within the Department of Surgery, Massachusetts General Hospital/ Harvard Medical School, Boston, Massachusetts; Shriners Hospitals for Children, Boston, Massachusetts.
Background:
Machine perfusion is gaining interest as an efficient method of tissue preservation of Vascularized Composite Allografts (VCA). The aim of this study was to develop a protocol for ex vivo subnormothermic oxygenated machine perfusion (SNMP) on rodent hindlimbs and to validate our protocol in a heterotopic hindlimb transplant model.
Methods:
In this optimization study we compared three different solutions during 6 h of SNMP (n = 4 per group). Ten control limbs were stored in a preservation solution on Static Cold Storage [SCS]). During SNMP we monitored arterial flowrate, lactate levels, and edema. After SNMP, muscle biopsies were taken for histology examination, and energy charge analysis. We validated the best perfusion protocol in a heterotopic limb transplantation model with 30-d follow up (n = 13). As controls, we transplanted untreated limbs (n = 5) and hindlimbs preserved with either 6 or 24 h of SCS (n = 4 and n = 5).
Results:
During SNMP, arterial outflow increased, and lactate clearance decreased in all groups. Total edema was significantly lower in the HBOC-201 group compared to the BSA group (P = 0.005), 4.9 (4.3-6.1) versus 48.8 (39.1-53.2) percentage, but not to the BSA + PEG group (P = 0.19). Energy charge levels of SCS controls decreased 4-fold compared to limbs perfused with acellular oxygen carrier HBOC-201, 0.10 (0.07-0.17) versus 0.46 (0.42-0.49) respectively (P = 0.002).
Conclusions:
Six hours ex vivo SNMP of rodent hindlimbs using an acellular oxygen carrier HBOC-201 results in superior tissue preservation compared to conventional SCS.
