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Metabolic Profiling of Skeletal Muscle During Ex-Vivo Normothermic Limb Perfusion
Elizabeth Rohde, Maryam Goudarzi PhD, Maria Madajka PhD, Carlos Ordenana MD, Majid Rezaei DDS, Sayf Al-deen Said MD, Vahe Fahradyan MD, Edoardo Dalla Pozza MD, Henri Brunengraber MD, Srinivasan Dasarathy MD, Antonio Rampazzo MD PhD, Bahar Bassiri Gharb MD PhD
Cleveland Clinic Plastic Surgery department Research Fellow
2019-02-14
Presenter: Carlos Ordenana
Affidavit:
I certify that the material proposed for presentation in this abstract has not been published in any scientific journal or previously presented at a major meeting
Director Name: Bassiri Bahar
Author Category: Fellow Plastic Surgery
Presentation Category: Basic Science Research
Abstract Category: General Reconstruction
Background: This study aimed to characterize the unique metabolic signature of skeletal muscle at 24h during ex-vivo normothermic human and porcine limb perfusion (NEVLP).
Methods: Six porcine forelimbs and seven human upper extremities undergoing NEVLP for over 24 hours were included. Limb muscle biopsies were collected at times 0, 6, 12, 24 hours of NEVLP or cold storage. Perturbations in the metabolomic signature of the limbs during NEVLP were determined via liquid chromatography–mass spectrometry (LC-MS)-based metabolomics. The LC-MS was operated in positive and negative electrospray ionization modes, scanning over a mass range of 50-750 Da at 120,000 resolution in full MS mode. The XCMS-extracted peak-table was evaluated using MetaboLyzer. The tandem MS spectra in this study were used for ion identification in Compound Discoverer 2.1 (Thermo) using the mzCloud spectral library.
Results: 39 ions and 58 ions with a putative ID at time point (TP) 24h were identified to be perturbed between the experimental and control groups in porcine forelimbs and human upper limbs respectively. Taurine and tryptophan were found to be the only common metabolites validated between human and porcine limbs at 24 hours. Common pathways at TP 24h included phenylalanine, tyrosine, and tryptophan biosynthesis, tryptophan degradation, and neuroactive ligand-receptor interaction.
Conclusions: Taurine and tryptophan could be indicative of muscle tissue breakdown and increased acidosis at TP 24h. The metabolomic profile of perfused muscles can be a new diagnostic tool to determine the effectiveness of long-term perfusion and to identify early biomarkers of muscle tissue degradation.
