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Peer-reviewedCC BY 4.0Artificial Organs

Monitoring Mitochondrial Function With High‐Resolution Respirometry During Ex‐Situ Heart Perfusion

  • Lukas Stastny1
  • Julia Hofmann2
  • Andras T. Meszaros2
  • Alexandra Ampferer1
  • Gabriel Putzer3
  • Nina Perkins1
  • Natalie Huemer1
  • Florian Sommerauer1
  • Judith Martini3
  • Julian Wagner3
  • Michael Grimm1
  • Stefan Schneeberger2
1 more authors
  • Julia Dumfarth1,4
4 affiliations
  1. 1Department of Cardiac Surgery Medical University of Innsbruck Innsbruck Austria
  2. 2Department of Visceral, Transplant and Thoracic Surgery Medical University of Innsbruck Innsbruck Austria
  3. 3Department of Anaesthesiology and Critical Care Medicine Medical University of Innsbruck Innsbruck Austria
  4. 4Department of Cardiac and Thoracic Aortic Surgery Medical University of Vienna Vienna Austria
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Abstract

Background: Due to the high adenosine‐triphosphate (ATP) consumption of the myocardial tissue, cardiac function is dependent on efficient mitochondrial respiration. Ischemia and reperfusion injury (I/R) has a severe impact on the function of the mitochondrial respiratory system. Ex‐situ heart perfusion (ESHP) can minimize I/R injury. We investigated mitochondrial respiration using high‐resolution respirometry (HRR) in a porcine model during ESHP.

Methods: Six German domestic pigs were used as heart and blood donors. Normothermic ESHP was performed over 6 h. Tissue homogenate of porcine myocardium was assessed by HRR at baseline (starting sample), after 60, 180, and 360 min of ESHP.

Results: The combined oxidative phosphorylation capacity with NADH, fatty acid and succinate fuel substrates remained stable during ESHP (starting sample: 198 ± 59.7, 60 min: 196.5 ± 66.7, 180 min: 207.5 ± 48.1, 360 min: 200.5 ± 27.9 pmol·s−1·mg wet mass−1; p = 0.707). The flux control ratio of the nicotinamide adenine dinucleotide hydrogen (NADH) pathway (starting sample: 0.43 ± 0.07, 60 min: 0.43 ± 0.05, 180 min: 0.47 ± 0.05, 360 min: 0.44 ± 0.07 pmol·s−1·mg wet mass−1; p = 0.549) and succinate‐pathway (starting sample: 0.49 ± 0.06, 60 min: 0.49 ± 0.05, 180 min: 0.48 ± 0.01, 360 min: 0.5 ± 0.05 pmol·s−1·mg wet mass−1; p = 0.814) did not change during perfusion. The mitochondrial outer membrane damage was significantly decreased after 360 min of ESHP (starting sample: 0.39 ± 0.07, 60 min: 0.27 ± 0.06, 180 min: 0.18 ± 0.03, 360 min: 0.17 ± 0.02; p = 0.0124).

Conclusion: Our findings demonstrate that normothermic ESHP preserves mitochondrial respiratory function in a porcine model over 6 h. These results provide a mechanistic basis for the potential protective role of normothermic ESHP against ischemia–reperfusion injury. HRR represents a promising investigational tool for in‐depth mitochondrial assessment during ESHP.

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Cite

BibTeXRIS
BibTeX
@article{stastny2026monitoring,
  title = {{Monitoring Mitochondrial Function With High‐Resolution Respirometry During Ex‐Situ Heart Perfusion}},
  author = {Lukas Stastny and Julia Hofmann and Andras T. Meszaros and Alexandra Ampferer and Gabriel Putzer and Nina Perkins and Natalie Huemer and Florian Sommerauer and Judith Martini and Julian Wagner and Michael Grimm and Stefan Schneeberger and Julia Dumfarth},
  journal = {Artificial Organs},
  year = {2026},
  month = sep,
  doi = {10.1111/aor.70213},
  url = {https://doi.org/10.1111/aor.70213},
  note = {CC BY 4.0},
}
RIS
TY  - JOUR
TI  - Monitoring Mitochondrial Function With High‐Resolution Respirometry During Ex‐Situ Heart Perfusion
AU  - Lukas Stastny
AU  - Julia Hofmann
AU  - Andras T. Meszaros
AU  - Alexandra Ampferer
AU  - Gabriel Putzer
AU  - Nina Perkins
AU  - Natalie Huemer
AU  - Florian Sommerauer
AU  - Judith Martini
AU  - Julian Wagner
AU  - Michael Grimm
AU  - Stefan Schneeberger
AU  - Julia Dumfarth
PY  - 2026
DA  - 2026/09/10
JO  - Artificial Organs
DO  - 10.1111/aor.70213
UR  - https://doi.org/10.1111/aor.70213
AB  - Background: Due to the high adenosine‐triphosphate (ATP) consumption of the myocardial tissue, cardiac function is dependent on efficient mitochondrial respiration. Ischemia and reperfusion injury (I/R) has a severe impact on the function of the mitochondrial respiratory system. Ex‐situ heart perfusion (ESHP) can minimize I/R injury. We investigated mitochondrial respiration using high‐resolution respirometry (HRR) in a porcine model during ESHP.  Methods: Six German domestic pigs were used as heart and blood donors. Normothermic ESHP was performed over 6 h. Tissue homogenate of porcine myocardium was assessed by HRR at baseline (starting sample), after 60, 180, and 360 min of ESHP.  Results: The combined oxidative phosphorylation capacity with NADH, fatty acid and succinate fuel substrates remained stable during ESHP (starting sample: 198 ± 59.7, 60 min: 196.5 ± 66.7, 180 min: 207.5 ± 48.1, 360 min: 200.5 ± 27.9 pmol·s−1·mg wet mass−1; p = 0.707). The flux control ratio of the nicotinamide adenine dinucleotide hydrogen (NADH) pathway (starting sample: 0.43 ± 0.07, 60 min: 0.43 ± 0.05, 180 min: 0.47 ± 0.05, 360 min: 0.44 ± 0.07 pmol·s−1·mg wet mass−1; p = 0.549) and succinate‐pathway (starting sample: 0.49 ± 0.06, 60 min: 0.49 ± 0.05, 180 min: 0.48 ± 0.01, 360 min: 0.5 ± 0.05 pmol·s−1·mg wet mass−1; p = 0.814) did not change during perfusion. The mitochondrial outer membrane damage was significantly decreased after 360 min of ESHP (starting sample: 0.39 ± 0.07, 60 min: 0.27 ± 0.06, 180 min: 0.18 ± 0.03, 360 min: 0.17 ± 0.02; p = 0.0124).  Conclusion: Our findings demonstrate that normothermic ESHP preserves mitochondrial respiratory function in a porcine model over 6 h. These results provide a mechanistic basis for the potential protective role of normothermic ESHP against ischemia–reperfusion injury. HRR represents a promising investigational tool for in‐depth mitochondrial assessment during ESHP.
SN  - 0160-564X
SN  - 1525-1594
ER  -