Prolonged Cold Ischemia Time Results in Local and Remote Organ Dysfunction in a Murine Model of Vascularized Composite Transplantation

Abstract

Vascularized composite allotransplantation (VCA) is a viable reconstructive option for complex tissue defects. Although grafts with a large muscular component may be uniquely susceptible to ischemia-reperfusion (I/R) syndrome, the safe cold ischemia time in VCA has not been established. We investigated the effects of cold ischemia on innate immune response and recipient survival in a murine orthotopic hindlimb transplantation model. Surprisingly, mice receiving grafts exposed to 6 h or longer of cold storage demonstrated reduced survival and massive elevations in serum creatinine, blood urea nitrogen, and creatine kinase, compared with 1 h of cold storage recipients. This was accompanied by progressive increase in macrophage and neutrophil cell infiltration in muscle biopsy specimens, altered platelet endothelial cell adhesion molecule-1 expression, and ultimate renal injury. Multiplex immunoassay analysis identified 21 cytokines in serum and 18 cytokines in muscle biopsy specimens that are likely essential in the complex response to I/R-triggered injury in VCA. In conclusion, this study, in a mouse model of orthotopic hindlimb transplantation, is the first to document that prolonged cold ischemia triggers progressive I/R injury with vascular endothelial damage and may lead to irrecoverable local and remote organ damage. These experimental findings are important in guiding future therapies for human VCA recipients.

Beneficial effects of the novel marine oxygen carrier M101 during cold preservation of rat and human pancreas

“…Additionally, it is known that ROS production induced by mitochondrial dysfunction leads to structural and functional cell damages . Two distinct phases of cold ischaemia were observed up until 6 hours in this study, and our results confirm observations in several other organs, such as the liver and kidney . We confirmed that beyond 6 hours, islets started to lose function.…”

“…38 Two distinct phases of cold ischaemia were observed up until 6 hours in this study, and our results confirm observations in several other organs, such as the liver and kidney. [39][40][41] We confirmed that beyond 6 hours, islets started to lose function. However, we cannot exclude that this effect might be due to the use of HBSS and further studies using preservation solution would strengthen the study.…”

Effect of Static Cold Storage on Skeletal Muscle after Vascularized Composite Tissue Allotransplantation

“…reperfusion phase, resulting in an increased incidence of acute and chronic rejection episodes, and organ failures. 3,4 In hand allografts, the critical time of cold ischemia resulting in permanent damage is currently unknown. According to the international hand transplantation registry, cold ischemia times ranged from 30 minutes to 13.5 hours (average 5.5 hours) in hand allografts.…”

“…This wide variation may be one of the factors explaining the difference in function and long-term survival of hand allografts. 3,5,6 In this study, we aimed to investigate the long-term metabolic and structural effects of SCS on limb allografts using a syngeneic rat hindlimb allograft model. A syngeneic model represents all characteristics of VCAs without the need for immunosuppression.…”

Composite tissue allotransplantation: opportunities and challenges

“…In a mouse orthotopic hindlimb transplant model, for example, prolonged ischemic times have been associated with diminished graft survival. 64 Notably, tolerable ischemic times have not been established for VCA grafts. Utilizing standard preservation methods, cold ischemic times up to 6 h have been cited as an upper threshold in VCAs.…”