A Novel Ex Vivo Energy-Enhanced Hypothermic Preservation of Skin-Containing Composite Tissues
Technical Report,15 Aug 2018,14 Aug 2019
University of Arizona Tucson United States
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The overall objective of this project was to increase composite tissue tolerance to 24 hours of extended cold ischemia time. To achieve this, paired hindlimbs in rats were randomized to receive isolated hypothermic perfusions with either the University of Wisconsin UW solution control, or the UW ATPv solution experimental. ATPv are nanoscale fusogenic lipid vesicles encapsulating ATP and capable of delivering ATP directly into the cytosol of cells. Upon completion of the perfusion, hindlimbs were conserved in static cold storage for either 12, 16 or 24 hours before skin and muscles were harvested for viability assessment. The extended cold ischemia times invariably degraded tissue nucleotides and countered the optimum temperature required for enzymatic actions. These effects rendered nucleotide- and enzyme-dependent tests of viability unreliable. Energy enhanced ATPv ex vivo hypothermic perfusions extended hindlimb composite tissue tolerance to 24h of cold ischemia without remarkable vascular or muscle damages. The vascular endothelium exhibited the least tolerance to cold ischemia relative to the skeletal muscle and skin. The addition of O2 to the crystalloid perfusion solutions does not enhance composite tissue tolerance to cold ischemia. Linking the subcellular structural changes to the prevailing proteome provided the best muscle viability index after cold ischemia.
- Medicine and Medical Research
- Anatomy and Physiology