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Metabolic profiling during ex vivo normothermic pig limb perfusion – a new assessment of preserved limbs.

Maria Madajka - Niemeyer, Jillian Krebs, Vahe Fahradyan, Edoardo Dalla Pozza, Bahar Bassiri Gharb
Cleveland Clinic
2018-02-14

Presenter: Maria Madajka-Niemeyer

Affidavit:
This is the original work of the presenter

Director Name: Bassiri-Gharb

Author Category: Fellow Plastic Surgery
Presentation Category: Basic Science Research
Abstract Category: Hand

Background: The goal of this study was to determine relative concentrations of metabolites accumulated in the skeletal muscles during the long term normothermic pig limb perfusion. Relative concentration changes of metabolites were determined in order to establish the impact of perfusion system on metabolic pathways of skeletal muscles.

Methods: Pig muscles biopsies were collected at time 0, 12 and 24h of normothermic perfusion and analyzed by mass spectrometry (mass/charge ratio: 50-1000m/z). Identification of metabolites was performed by METLIN software based on molecular masses, retention time and peak area intensity.

Results: A total of over 4,000 molecules were obtained from all the samples. Final analysis was performed for those with analytical significance (metabolite mass/blank sample control ratio >5) which excluded ROS.

Metabolic profiles changed rapidly during 24 hour perfusion. A total of 54 peptides, 28 esters, 48 ketones and over 20 acids, alcohols and amines were identified. Aldehyde contained 66.6% of all metabolites identified at time 0 of perfusion. In contrast, at time point 12 hours sugars content dominated the pool of metabolites at 61.5% with the decrease to 15% at 24 hours. At the end of perfusion both: peptides and esters were the dominant chemical group at 55% and 57%, respectively. High content of peptides at the end of perfusion might indicate muscle damage due to ischemic changes.

Conclusion: 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 ischemia.

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