Hypothermic Machine Perfusion of Composite Tissues

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Virginia Tech


Organ perfusion systems have successfully been applied to solid organ preservation and subsequent transplantation. However, their use in limb preservation for Vascularized Composite Tissue Allotransplantation (VCA) has yet to be thoroughly investigated. This thesis explores the potential for hypothermic machine perfusion in prolonging limb graft viability in a swine forelimb amputation model. The experiment was designed with the right and left forelimbs from the same pig randomly assigned to the treatment and control groups. Eighteen (18) limbs were procured from a local abattoir, vessels cannulated, and an initial flush of a modified phosphate buffered saline (PBS) solution was performed. Half of those limbs, assigned to the treatment group, were then preserved with continuous hypothermic machine perfusion for 12 hours. The perfusate was a PBS solution supplemented with 5% w/v dextrose. The remaining nine limbs, assigned to the control group, were placed into a plastic bag and kept at room temperature (ca. 20oC) for the entire duration of the experiment. Methylene blue was used to verify perfusion throughout limbs. Histopathological analysis revealed the presence of significantly greater deterioration of the perfused limbs compared to control. I concluded that PBS solution is not suitable for extended limb preservation. Inadequate perfusate volume and lack of solution replenishment resulted in metabolic waste build up, accelerating total organ damage. Continued research is needed in order to develop clinically relevant hypothermic machine perfusion devices capable of prolonged limb preservation.



Engineering, Mechanical Engineering, Biomedical