Ultra-weak photon emission as a dynamic tool for monitoring oxidative stress metabolism

Rosilene Cristina Rossetto Burgos,Rawi Ramautar,Michal Cifra,Jan Van Der Greef,Lennart Jan Van Winden,Eduard P A Van Wijk,Thomas Hankemeier,Ruud Berger,Johannes Cornelius Schoeman,Kateřina Červinková

doi:10.1038/s41598-017-01229-x

Abstract

In recent years, excessive oxidative metabolism has been reported as a critical determinant of pathogenicity in many diseases. The advent of a simple tool that can provide a physiological readout of oxidative stress would be a major step towards monitoring this dynamic process in biological systems, while also improving our understanding of this process. Ultra-weak photon emission (UPE) has been proposed as a potential tool for measuring oxidative processes due to the association between UPE and reactive oxygen species. Here, we used HL-60 cells as an in vitro model to test the potential of using UPE as readout for dynamically monitoring oxidative stress after inducing respiratory burst. In addition, to probe for possible changes in oxidative metabolism, we performed targeted metabolomics on cell extracts and culture medium. Lastly, we tested the effects of treating cells with the NADPH oxidase inhibitor diphenyleneiodonium chloride (DPI). Our results show that UPE can be used as readout for measuring oxidative stress metabolism and related processes.

Highlights

In recent years, excessive oxidative metabolism has been reported as a critical determinant of pathogenicity in many diseases
reactive oxygen species (ROS) are produced by mitochondria as a product of cellular metabolism[4]; during respiratory burst NADPH oxidase plays a central role in ROS production for cellular defence[5, 6]
HL-60 cells were differentiated into neutrophil-like cells by incubation in all-trans retinoic acid (ATRA) for up to 7 days

Summary

Introduction

Excessive oxidative metabolism has been reported as a critical determinant of pathogenicity in many diseases. We used HL-60 cells as an in vitro model to test the potential of using UPE as readout for dynamically monitoring oxidative stress after inducing respiratory burst. ROS production has been analysed using various techniques, including photometry, luminometry, flow cytometry, and precipitation reactions[17] All of these techniques provide a measure at only a single time point or require labels; these techniques are cell-dependent, laborious, and not necessarily feasible for diagnostic purposes. UPE is a potential new tool for monitoring dynamic biological processes that involve ROS, including ROS-related diseases[24, 25], as well as processes related to oxidative stress metabolism. A clear advantage of UPE is that it provides spatiotemporal information; in addition, UPE is non-damaging, non-invasive, label-free, and relatively cost-effective

Methods

Results

Conclusion