Extra-corporeal in-vitro perfusion of isolated skeletal muscle flaps improves ischaemic survival

MMed thesis - Faculty of Health SciencesThe field of organ and tissue transplantation has necessitated an improved understanding of their associated pathophysiological pathways. Specific areas of interest involve the changes that follow ischaemia and derangement’s that accompany organ and tissue storage, reperfusion injury and the “no-reflow” phenomenon. Strategies have been devised to manipulate and modify these processes, improving tissue and organ survival and function. These have involved the use of preservation solutions. Although most research involves organ transplantation, these principles have been translated and applied to various tissues, surgical flaps and microvascular replantations. These studies have generally used the skin flap as their model with little knowledge regarding muscle flaps, the most vulnerable to the ischaemic process. This study targets the use of one such preservation system and uses skeletal muscle as its tissue model. The vascular anatomy of the rectus femoris muscle in the New Zealand white rabbit was studied anatomically and radiologically and thus described. The isolated rectus femoris muscle flap was harvested and perfused in-vitro with cooled, oxygenated University of Wisconsin solution (UWS) using a pulsatile renal perfusion pump. UWS was selected as it contains vital additives important in cryopreservation of organs. Monitoring of various physiological parameters was performed. The muscle was examined at 0, 4, 8, 12, 18 and 24 hours of extra-corporeal perfusion using warm and cold, non-perfused controls. The contralateral muscle served as the control. End-points were the percentage of muscle survival, as determined by a new grading system of muscle ischaemia, based on 3 light and 7 electron microscopic criteria. The overall percentage of muscle survival (combined light and electron microscopy scores) resulted in approximately 58% survival at 24 hours for the perfused muscle versus 31% for the cold stored muscle. The stored muscle had the same survival rate at 12 hours as did the perfused muscle at 24 hours. For all time periods beyond 4 to 8 hours, perfused muscle showed statistically improved survival rates compared to the stored muscle. Eight hours appears to be a crucial point beyond which survival in muscle deteriorates to a much greater degree without perfusion. Questions remain as to which method of preservation yields the best survival benefit and, as yet, there is no “ideal” perfusate. The future involves manipulating perfusion solutions and trying to arrest or reverse established warm ischaemia. Success of free tissue transfers and replantations of musclecontaining body parts may be enhanced. These techniques may also allow us to effectively store previously harvested flaps and eventually, to enter the realm of “banked” allograft tissue flaps