Frontiers in Molecular Biosciences (May 2024)

The impact of normothermic and hypothermic preservation methods on kidney lipidome—comparative study using chemical biopsy with microextraction probes

  • Natalia Warmuzińska,
  • Kamil Łuczykowski,
  • Iga Stryjak,
  • Hernando Rosales-Solano,
  • Peter Urbanellis,
  • Janusz Pawliszyn,
  • Markus Selzner,
  • Markus Selzner,
  • Barbara Bojko

DOI
https://doi.org/10.3389/fmolb.2024.1341108
Journal volume & issue
Vol. 11

Abstract

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IntroductionNormothermic ex vivo kidney perfusion (NEVKP) is designed to replicate physiological conditions to improve graft outcomes. A comparison of the impact of hypothermic and normothermic preservation techniques on graft quality was performed by lipidomic profiling using solid-phase microextraction (SPME) chemical biopsy as a minimally invasive sampling approach.MethodsDirect kidney sampling was conducted using SPME probes coated with a mixed-mode extraction phase in a porcine autotransplantation model of the renal donor after cardiac death, comparing three preservation methods: static cold storage (SCS), NEVKP, and hypothermic machine perfusion (HMP). The lipidomic analysis was done using ultra-high-performance liquid chromatography coupled with a Q-Exactive Focus Orbitrap mass spectrometer.ResultsChemometric analysis showed that the NEVLP group was separated from SCS and HMP groups. Further in-depth analyses indicated significantly (p < 0.05, VIP > 1) higher levels of acylcarnitines, phosphocholines, ether-linked and longer-chain phosphoethanolamines, triacylglycerols and most lysophosphocholines and lysophosphoethanolamines in the hypothermic preservation group. The results showed that the preservation temperature has a more significant impact on the lipidomic profile of the kidney than the preservation method’s mechanical characteristics.ConclusionHigher levels of lipids detected in the hypothermic preservation group may be related to ischemia-reperfusion injury, mitochondrial dysfunction, pro-inflammatory effect, and oxidative stress. Obtained results suggest the NEVKP method’s beneficial effect on graft function and confirm that SPME chemical biopsy enables low-invasive and repeated sampling of the same tissue, allowing tracking alterations in the graft throughout the entire transplantation procedure.

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