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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

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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 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.

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(Adapted from Fontana 2001)

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(Adapted from Prawira 2010)

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Acknowledgements

Firstly, our utmost gratitude and respect for the generosity of the tissue donors and their families. Furthermore, this work was made possible through the collaborative effort involving a number of Australian research hubs, institutions and individuals in response to an invitation of the International Atomic Energy Agency (Project: CRP E31006). Their scientific input and in-kind contributions are acknowledged: Victorian Institute of Forensic Medicine—Donor Tissue Bank of Victoria (in particular to tissue donor coordinators, retrieval and processing teams; and for access to processing facility and equipment; Histopathology Dept., in particular David Cauchi for the preparation and staining of samples); Monash University—Monash Micro Imaging (in particular, Stephen Cody for the provision of access to bright field instrumentation, training and general imaging input); Monash Histology Platform (in particular Camilla Cohen and Jonathan Bensley, for advise on Picrosirius red staining); Melbourne University—Department of Chemical and Biomolecular Engineering Post Graduate Studies Program (in particular Honors students Eleanor Lazarus and Sunny L Prawira). Last but certainly not least, this work pays a tribute to our Professor, Glynn Phillips. His enthusiasm, dedication and generosity has inspired a generation of budding tissue bankers worldwide including myself, the main author; his moto “my bone is your bone”—i.e. sharing knowledge is essential—resonates in all of us. His unrelenting positive attitude has steered my path in the wonderful but challenging world of tissue donation, banking and transplantation. For the great moments and teachings, my sincere gratitude.

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Authors and Affiliations

  1. Department of Surgery - Central Medical School, Monash University, Melbourne, Australia

    M. R. Herson

  2. Donor Tissue Bank of Victoria - Victorian Institute of Forensic Medicine, Melbourne, Australia

    K. Hamilton & S. Poniatowski

  3. CSIRO - Manufacturing, Clayton, Australia

    J. White

  4. CSIRO - Data 61, Clayton, Australia

    D. Alexander

  5. Department of Chemical and Biomolecular Engineering, University of Melbourne, Melbourne, Australia

    A. J. O’Connor

  6. Hudson Institute of Medical Research, Monash University, Clayton, Australia

    J. A. Werkmeister

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  1. M. R. Herson
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  2. K. Hamilton
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  3. J. White
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  4. D. Alexander
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  5. S. Poniatowski
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  6. A. J. O’Connor
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  7. J. A. Werkmeister
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Correspondence to M. R. Herson.

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Human and animal rights

This article does not contain studies with human participants or animals by any of the authors. Human donated tissues were made available by the Donor Tissue Bank of Victoria (DTBV), following approval by the Ethics Committee of the Victorian Institute of Forensic Medicine (VIFM EC15/2009). Release of tissues for improving in house processes and technique was in accordance with family informed consent.

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Herson, M.R., Hamilton, K., White, J. et al. 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. Cell Tissue Bank 19, 215–227 (2018). https://doi.org/10.1007/s10561-018-9694-1

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  • DOI: https://doi.org/10.1007/s10561-018-9694-1

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