Real-time monitoring of mitochondrial oxygenation during machine perfusion using resonance Raman spectroscopy predicts organ function

Theodor Bilharz Research Institute

Bibliographic Information

Authors: Jain R.; Ajenu E.O.; Lopera Higuita M.; Hafiz E.O.A.; Muzikansky A.; Romfh P.; Tessier S.N.

Journal: Scientific Reports

Publisher: Nature Research

Publication Date: 27 March 2024

Volume / Issue: Volume 14 / Issue 1

Article No.: 7328

ISSN: 20452322

DOI: 10.1038/s41598-024-57773-w

Scopus: View on Scopus

PubMed: 38538723

Document Type: Article

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

Authors and Affiliations

Jain R., Center for Engineering in Medicine and Surgery, Harvard Medical School and Massachusetts General Hospital, Boston, MA, United States, Shriners Children’s Hospital, Boston, MA, United States; Ajenu E.O., Center for Engineering in Medicine and Surgery, Harvard Medical School and Massachusetts General Hospital, Boston, MA, United States, Shriners Children’s Hospital, Boston, MA, United States; Lopera Higuita M., Center for Engineering in Medicine and Surgery, Harvard Medical School and Massachusetts General Hospital, Boston, MA, United States, Shriners Children’s Hospital, Boston, MA, United States; Hafiz E.O.A., Department of Electron Microscopy Research, Clinical Laboratory Division, Theodor Bilharz Research Institute, Giza, Egypt; Muzikansky A., Biostatistics Center, Massachusetts General Hospital, Boston, MA, United States; Romfh P., Pendar Technologies, Cambridge, MA, United States; Tessier S.N., Center for Engineering in Medicine and Surgery, Harvard Medical School and Massachusetts General Hospital, Boston, MA, United States, Shriners Children’s Hospital, Boston, MA, United States

Abstract

Organ transplantation is a life-saving procedure affecting over 100,000 people on the transplant waitlist. Ischemia reperfusion injury (IRI) is a major challenge in the field as it can cause post-transplantation complications and limit the use of organs from extended criteria donors. Machine perfusion technology has the potential to mitigate IRI; however, it currently fails to achieve its full potential due to a lack of highly sensitive and specific assays to assess organ quality during perfusion. We developed a real-time and non-invasive method of assessing organs during perfusion based on mitochondrial function and injury using resonance Raman spectroscopy. It uses a 441 nm laser and a high-resolution spectrometer to quantify the oxidation state of mitochondrial cytochromes during perfusion. This index of mitochondrial oxidation, or 3RMR, was used to understand differences in mitochondrial recovery of cold ischemic rodent livers during machine perfusion at normothermic temperatures with an acellular versus cellular perfusate. Measurement of the mitochondrial oxidation revealed that there was no difference in 3RMR of fresh livers as a function of normothermic perfusion when comparing acellular versus cellular-based perfusates. However, following 24 h of static cold storage, 3RMR returned to baseline faster with a cellular-based perfusate, yet 3RMR progressively increased during perfusion, indicating injury may develop over time. Thus, this study emphasizes the need for further refinement of a reoxygenation strategy during normothermic machine perfusion that considers cold ischemia durations, gradual recovery/rewarming, and risk of hemolysis. © The Author(s) 2024.

Keywords

Humans; Liver; Liver Transplantation; Mitochondria; Organ Preservation; Perfusion; Spectrum Analysis, Raman; human; metabolism; mitochondrion; procedures; Raman spectrometry

Citation Information

Scopus Citations: 9

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