The measurement of water activity in allogeneic skin grafts preserved using high concentration glycerol or propylene glycol.
In the presence of free water, many degradation reactions can occur within stored tissues including enzymatic digestion, oxidation (peroxidation) and hydrolytic reactions, as well as the detrimental effects of microbial growth, therefore most long-term banking techniques are designed to avoid free water. One method currently used for banking of skin grafts is the use of high concentration (85%) glycerol as a preservative. In this case, the glycerol was assumed to dehydrate the skin by osmosis and diffusion out of the cells and skin matrix respectively. We have recently shown that this assumption is incorrect and the converse occurs, i.e. glycerol enters the skin and sequesters the water. It was therefore essential to determine whether enough water had been immobilised to prevent degradation of the tissue. Using an instrument (Pawkit) designed to measure water activity (aw) it was shown that a stepwise reduction in aw was achieved when the skin was immersed in 50 and 85% glycerol or propylene glycol, respectively. At the end of the glycerolisation process, the final aw was shown to be circa 0.3. An aw of 0.3 is known to minimise lipid peroxidation and reduce other degradation reaction rates to very low levels. It was concluded that the current glycerolisation protocol results in effective sequestration of water avoiding degradation of the skin during storage. The method presented should be used as a quality control step to confirm adequacy of preservation for each batch of glycerolised skin.
Ross
Alexandra
A
Tissue Services R&D, National Blood Service, Longley Lane, Sheffield S5 7JN, UK.
Kearney
John N
JN
D002102:Cadaver; D002470:Cell Survival; D015925:Cryopreservation / Q000379:methods; D003451:Cryoprotective Agents / Q000737:chemistry; D005990:Glycerol / Q000737:chemistry*; D006801:Humans; D066298:In Vitro Techniques; D019946:Propylene Glycol / Q000737:chemistry*; D012867:Skin / Q000737:chemistry* / Q000473:pathology*; D016038:Skin Transplantation / Q000379:methods*; D014015:Tissue Banks; D014021:Tissue Preservation / Q000379:methods*; D014184:Transplantation, Homologous / Q000379:methods; D014867:Water / Q000032:analysis* / Q000737:chemistry
D003451:Cryoprotective Agents; D014867:Water; D019946:Propylene Glycol; D005990:Glycerol