Optical Spectroscopic Detection of Mitochondrial Biomarkers (FMN and NADH) for Hypothermic Oxygenated Machine Perfusion: A Comparative Study in Different Perfusion Media

Optical Spectroscopic Detection of Mitochondrial Biomarkers (FMN and NADH) for Hypothermic Oxygenated Machine Perfusion: A Comparative Study in Different Perfusion Media

Ex situ machine perfusion has emerged as a pivotal technique for organ preservation and pre-transplant viability assessment, where the real-time monitoring of mitochondrial biomarkers—flavin mononucleotide (FMN) and nicotinamide adenine dinucleotide (NADH)—could significantly mitigate ischemia-reper...

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Main Authors: Lorenzo Agostino Cadinu, Keyue Sun, Chunbao Jiao, Rebecca Panconesi, Sangeeta Satish, Fatma Selin Yildirim, Omer Faruk Karakaya, Chase J. Wehrle, Geofia Shaina Crasta, Fernanda Walsh Fernandes, Nasim Eshraghi, Koki Takase, Hiroshi Horie, Pier Carlo Ricci, Davide Bagnoli, Mauricio Flores Carvalho, Andrea Schlegel, Massimo Barbaro
Format: Article
Language:English
Published: MDPI AG 2025-06-01
Series:Sensors
Subjects:
mitochondrial molecular biomarkers
spectroscopy application
optic phenomena in perfusion solution
spectroscopic calibration
methodologies and protocols
optic sensor theoretical foundations
Online Access:https://www.mdpi.com/1424-8220/25/13/4031
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Summary:Ex situ machine perfusion has emerged as a pivotal technique for organ preservation and pre-transplant viability assessment, where the real-time monitoring of mitochondrial biomarkers—flavin mononucleotide (FMN) and nicotinamide adenine dinucleotide (NADH)—could significantly mitigate ischemia-reperfusion injury risks. This study develops a non-invasive optical method combining fluorescence and UV-visible spectrophotometry to quantify FMN and NADH in hypothermic oxygenated perfusion media. Calibration curves revealed linear responses for both biomarkers in absorption and fluorescence (FMN: λ<sub>ex</sub> = 445 nm, λ<sub>em</sub> = 530–540 nm; NADH: λ<sub>ex</sub> = 340 nm, λ<sub>em</sub> = 465 nm) at concentrations < 100 μg mL<sup>−1</sup>. However, NADH exhibited nonlinear fluorescence above 100 μg mL<sup>−1</sup>, requiring shifted excitation to 365 nm for reliable detection. Spectroscopic analysis further demonstrated how perfusion solution composition alters FMN/NADH fluorescence properties, with consistent reproducibility across media. The method’s robustness was validated through comparative studies in clinically relevant solutions, proposing a strategy for precise biomarker quantification without invasive sampling. These findings establish a foundation for real-time, optical biosensor development to enhance organ perfusion monitoring. By bridging spectroscopic principles with clinical needs, this work advances translational sensor technologies for transplant medicine, offering a template for future device integration.
ISSN:1424-8220

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