Abstract

.Significance: Decreasing the oxygen consumption rate (OCR) of tumor cells is a powerful method for ameliorating tumor hypoxia. However, quantifying the change in OCR is challenging in complex experimental systems.Aim: We present a method for quantifying the OCR of two tumor cell lines using oxygen-sensitive dual-emissive boron nanoparticles (BNPs). We hypothesize that our BNP results are equivalent to the standard Seahorse assay.Approach: We quantified the spectral emissions of the BNP and accounted for external oxygen diffusion to quantify OCR over 24 h. The BNP-computed OCR of two breast cancer cell lines, E0771 and 4T07, were compared with their respective Seahorse assays. Both cell lines were also irradiated to quantify radiation-induced changes in the OCR.Results: Using a Bland–Altman analysis, our BNPs OCR was equivalent to the standard Seahorse assay. Moreover, in an additional experiment in which we irradiated the cells at their 50% survival fraction, the BNPs were sensitive enough to quantify 24% reduction in OCR after irradiation.Conclusions: Our results conclude that the BNPs are a viable alternative to the Seahorse assay for quantifying the OCR in cells. The Bland–Altman analysis showed that these two methods result in equivalent OCR measurements. Future studies will extend the OCR measurements to complex systems including 3D cultures and in vivo models, in which OCR measurements cannot currently be made.

Highlights

  • The oxygen consumption rate (OCR) provides quantitative information regarding cellular metabolism and the utilization of oxygen

  • Using a Bland–Altman analysis, our boron nanoparticles (BNPs) OCR was equivalent to the standard Seahorse assay

  • In an additional experiment in which we irradiated the cells at their 50% survival fraction, the BNPs were sensitive enough to quantify 24% reduction in OCR after irradiation

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Summary

Introduction

The oxygen consumption rate (OCR) provides quantitative information regarding cellular metabolism and the utilization of oxygen. One possibility for ameliorating the problem of hypoxia is to decrease the OCR of tumor cells;[2,3,4,5] Secomb et al,[6] using a mathematical model of tumor blood vessels, predicted that a decrease of 30% in the OCR of tumor cells would abolish hypoxia; compared with increasing oxygen delivery, decreasing the OCR could be 30× more efficient. To use this therapy, a reliable, accurate method for measuring the OCR both in vitro and in vivo is necessary

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