Interaction of preservation methods and radiation sterilization in human skin processing, with particular insight on the
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Interaction of preservation methods and radiation sterilization in human skin processing, with particular insight on the impact of the final water content and collagen disruption. Part I: process validation, water activity and collagen changes in tissues cryopreserved or processed using 50, 85 or 98% glycerol solutions M. R. Herson . K. Hamilton . J. White . D. Alexander . S. Poniatowski . A. J. O’Connor . J. A. Werkmeister Received: 9 December 2017 / Accepted: 17 April 2018 / Published online: 25 April 2018 Ó Springer Science+Business Media B.V., part of Springer Nature 2018
Abstract Current regulatory requirements demand an in-depth understanding and validation of protocols used in tissue banking. The aim of this work was to characterize the quality of split thickness skin allografts cryopreserved or manufactured using highly concentrated solutions of glycerol (50, 85 or 98%), where tissue water activity (aw), histology and birefringence changes were chosen as parameters. Consistent aw outcomes validated the proposed processing protocols. While no significant changes in tissue M. R. Herson (&) Department of Surgery - Central Medical School, Monash University, Melbourne, Australia e-mail: [email protected] K. Hamilton S. Poniatowski Donor Tissue Bank of Victoria - Victorian Institute of Forensic Medicine, Melbourne, Australia J. White CSIRO - Manufacturing, Clayton, Australia D. Alexander CSIRO - Data 61, Clayton, Australia A. J. O’Connor Department of Chemical and Biomolecular Engineering, University of Melbourne, Melbourne, Australia J. A. Werkmeister Hudson Institute of Medical Research, Monash University, Clayton, Australia
quality were observed under bright-field microscopy or in collagen birefringence, in-process findings can be harnessed to fine-tune and optimize manufacturing outcomes in particular when further radiation sterilization is considered. Furthermore, exposing the tissues to 85% glycerol seems to derive the most efficient outcomes as far as aw and control of microbiological growth. Keywords Skin allografts Processing Glycerol Water activity Birefringence
Introduction Banked human skin allografts have long since been recognized as the gold standard in temporary skin substitution, for example in large burns (Brown et al. 1953). Processing and long term storage of donated tissues under validated conditions safeguard their performance as skin substitutes comparable to autologous skin; until recognized by the recipient’s immunological system as foreign materials, engraftment will occur with the re-establishment of the natural barriers lost to trauma (Kagan 1998). More recently, the potential for definitive integration of allogeneic dermal collagen into the wound bed has been recognized, introducing the concept of
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transplanted dermis as an extracellular collagen matrix scaffold (Cuono et al. 1987; Schiozer et al. 1994). The initial use of split thickness skin grafts as the dermal source has been surpassed by the demonstrated effectiveness of processed and decellularized de
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