Real-time respirometry of isolated mitochondria during low irradiance laser exposure

Nathaniel J. Pope; Gary D. Noojin; Michael L. Denton

doi:10.1117/12.2608879

4 March 2022 Real-time respirometry of isolated mitochondria during low irradiance laser exposure

Nathaniel J. Pope, Gary D. Noojin, Michael L. Denton

Proceedings Volume 11940, Optical Methods for Tumor Treatment and Detection: Mechanisms and Techniques in Photodynamic and Photobiomodulation Therapy XXX; 119400C (2022) https://doi.org/10.1117/12.2608879
Event: SPIE BiOS, 2022, San Francisco, California, United States

Abstract

Photobiomodulation therapy (PBMT) is a promising but poorly understood technique utilizing low irradiance LED or laser light to induce positive biological effects. Though the use of PBMT in clinical settings is increasingly accepted, the molecular mechanisms underlying these effects remain poorly characterized. The most commonly postulated mechanism is via light absorption at cytochrome c oxidase, also known as complex IV (C-IV) of the mitochondrial electron transport chain. It is hypothesized that light absorption at C-IV initiates a cascade of events subsequently resulting in increased ATP generation, and improved mitochondrial/cellular fitness. However, surprisingly little is known about the mechanisms by which these pathways are initiated, and how they establish the observed beneficial outcomes. Other work from our group has demonstrated that the kinetics of C-IV enzyme activity can be positively modulated by lowirradiance NIR laser exposure. However, the nature of those assays limits them to a single post-exposure measurement of one enzyme complex. To extend this observation we have developed a novel method allowing us to conduct real-time measurements of mitochondrial respiration while simultaneously applying low irradiance laser exposures consistent with those used in photobiomodulation. This technique utilized a polarographic oxygen sensor to make continuous in vitro measurements of oxygen consumption, before, during, and after laser irradiation. Using an integrated optical port, laser irradiation was applied to the sample accurately and reliably, allowing for precision dosimetry. This novel technique was then used to probe effects of light exposure at a number of wavelengths and irradiances on mitochondrial respiration.

Conference Presentation

Citation Download Citation

Nathaniel J. Pope, Gary D. Noojin, and Michael L. Denton "Real-time respirometry of isolated mitochondria during low irradiance laser exposure", Proc. SPIE 11940, Optical Methods for Tumor Treatment and Detection: Mechanisms and Techniques in Photodynamic and Photobiomodulation Therapy XXX, 119400C (4 March 2022); https://doi.org/10.1117/12.2608879

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