The role of antifreeze glycoprotein (AFGP) and polyvinyl alcohol/polyglycerol (X/Z-1000) as ice modulators during partial freezing of rat livers

Theodor Bilharz Research Institute

Bibliographic Information

Authors: Tessier S.N.; Haque O.; Pendexter C.A.; Cronin S.E.J.; Hafiz E.O.A.; Weng L.; Yeh H.; Markmann J.F.; Taylor M.J.; Fahy G.M.; Toner M.; Uygun K.

Journal: Frontiers in Physics

Publisher: Frontiers Media SA

Publication Date: 22 December 2022

Volume / Issue: Volume 10

Article No.: 1033613

ISSN: 2296424X

DOI: 10.3389/fphy.2022.1033613

Scopus: View on Scopus

Document Type: Article

Access: All Open Access; Gold Open Access; Green Open Access

Authors and Affiliations

Tessier S.N., Center for Engineering in Medicine and Surgery, Massachusetts General Hospital, Harvard Medical School, Boston, MA, United States, Shriners Hospitals for Children, Boston, MA, United States; Haque O., Center for Engineering in Medicine and Surgery, Massachusetts General Hospital, Harvard Medical School, Boston, MA, United States, Shriners Hospitals for Children, Boston, MA, United States, Department of Surgery, Beth Israel Deaconess Medical Center, Boston, MA, United States, Department of Surgery, Division of Transplantation, Massachusetts General Hospital, Harvard Medical School, Boston, MA, United States; Pendexter C.A., Center for Engineering in Medicine and Surgery, Massachusetts General Hospital, Harvard Medical School, Boston, MA, United States, Shriners Hospitals for Children, Boston, MA, United States; Cronin S.E.J., Center for Engineering in Medicine and Surgery, Massachusetts General Hospital, Harvard Medical School, Boston, MA, United States, Shriners Hospitals for Children, Boston, MA, United States; Hafiz E.O.A., Department of Electron Microscopy Research, Theodor Bilharz Research Institute, Giza, Egypt; Weng L., Center for Engineering in Medicine and Surgery, Massachusetts General Hospital, Harvard Medical School, Boston, MA, United States, Shriners Hospitals for Children, Boston, MA, United States; Yeh H., Department of Surgery, Division of Transplantation, Massachusetts General Hospital, Harvard Medical School, Boston, MA, United States; Markmann J.F., Department of Surgery, Division of Transplantation, Massachusetts General Hospital, Harvard Medical School, Boston, MA, United States; Taylor M.J., Sylvatica Biotech Inc, North Charleston, SC, United States, Department of Mechanical Engineering, Carnegie Mellon University, Pittsburgh, PA, United States; Fahy G.M., 21st Century Medicine, Fontana, CA, United States; Toner M., Center for Engineering in Medicine and Surgery, Massachusetts General Hospital, Harvard Medical School, Boston, MA, United States, Shriners Hospitals for Children, Boston, MA, United States; Uygun K., Center for Engineering in Medicine and Surgery, Massachusetts General Hospital, Harvard Medical School, Boston, MA, United States, Shriners Hospitals for Children, Boston, MA, United States

Abstract

Introduction: The current liver organ shortage has pushed the field of transplantation to develop new methods to prolong the preservation time of livers from the current clinical standard of static cold storage. Our approach, termed partial freezing, aims to induce a thermodynamically stable frozen state at high subzero storage temperatures (−10°C to −15°C), while simultaneously maintaining a sufficient unfrozen fraction to limit ice-mediated injury. Methods and results: Using glycerol as the main permeating cryoprotectant agent, this research first demonstrated that partially frozen rat livers showed similar outcomes after thawing from either −10°C or −15°C with respect to subnormothermic machine perfusion metrics. Next, we assessed the effect of adding ice modulators, including antifreeze glycoprotein (AFGP) or a polyvinyl alcohol/polyglycerol combination (X/Z-1000), on the viability and structural integrity of partially frozen rat livers compared to glycerol-only control livers. Results showed that AFGP livers had high levels of ATP and the least edema but suffered from significant endothelial cell damage. X/Z-1000 livers had the highest levels of ATP and energy charge (EC) but also demonstrated endothelial damage and post-thaw edema. Glycerol-only control livers exhibited the least DNA damage on Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining but also had the lowest levels of ATP and EC. Discussion: Further research is necessary to optimize the ideal ice modulator cocktail for our partial-freezing protocol. Modifications to cryoprotective agent (CPA) combinations, including testing additional ice modulators, can help improve the viability of these partially frozen organs. Copyright © 2022 Tessier, Haque, Pendexter, Cronin, Hafiz, Weng, Yeh, Markmann, Taylor, Fahy, Toner and Uygun.

Keywords

antifreeze glycoprotein; cryopreservation; machine perfusion; partial freezing; polyglycerol; polyvinyl alcohol; Cold storage; Endothelial cells; Freezing; Glycerol; Glycoproteins; Ice; Rats; Thawing; 'current; Cryo-preservation; Energy; Frozen state; Polyglycerols; Rat livers; Thermodynamically stable; Polyvinyl alcohols

Citation Information

Scopus Citations: 12

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