Duke University, Department of Radiation Oncology, Duke University Medical Center, Durham, North Car, United States.
University of Virginia, Department of Chemistry, Charlottesville, Virginia, United States.
J Biomed Opt. 2020 Nov;25(11). doi: 10.1117/1.JBO.25.11.116504.
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.
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.
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.
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.
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.
降低肿瘤细胞的耗氧率（OCR）是改善肿瘤缺氧的有效方法。然而，在复杂的实验系统中，量化 OCR 的变化具有挑战性。
我们提出了一种使用氧敏双发射硼纳米粒子（BNP）定量两种肿瘤细胞系 OCR 的方法。我们假设我们的 BNP 结果与标准 Seahorse 测定等效。
我们量化了 BNP 的光谱发射，并考虑了外部氧气扩散，以在 24 小时内定量 OCR。将两种乳腺癌细胞系 E0771 和 4T07 的 BNP 计算的 OCR 与其各自的 Seahorse 测定进行了比较。还对这两种细胞系进行了辐照，以量化辐射诱导的 OCR 变化。
使用 Bland-Altman 分析，我们的 BNP OCR 与标准 Seahorse 测定等效。此外，在另一个实验中，我们在细胞的 50%存活分数时对其进行辐照，BNP 足够灵敏，可以在辐照后定量 OCR 降低 24%。
我们的结果得出结论，BNP 是 Seahorse 测定法在细胞中定量 OCR 的可行替代品。Bland-Altman 分析表明，这两种方法得出的 OCR 测量结果等效。未来的研究将把 OCR 测量扩展到包括 3D 培养物和体内模型在内的复杂系统，目前无法在这些系统中进行 OCR 测量。
