4.7 Article

A Novel Ruthenium-based Molecular Sensor to Detect Endothelial Nitric Oxide

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SCIENTIFIC REPORTS
卷 9, 期 -, 页码 -

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NATURE PUBLISHING GROUP
DOI: 10.1038/s41598-019-39123-3

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  1. Centre of excellence for Nanoscale BioPhotonics, through the Australian Research Council (ARC) [CE140100003]
  2. National Health and Medical Research Council (NHMRC) of Australia [1111630]
  3. Future Leader Fellowship from the National Heart Foundation (NHF) of Australia [FLF100412]
  4. National Health and Medical Research Council of Australia [1111630] Funding Source: NHMRC

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Nitric oxide (NO) is a key regulator of endothelial cell and vascular function. The direct measurement of NO is challenging due to its short half-life, and as such surrogate measurements are typically used to approximate its relative concentrations. Here we demonstrate that ruthenium-based [Ru(bpy)(2)(dabpy)](2+) is a potent sensor for NO in its irreversible, NO-bound active form, [Ru(bpy)(2)(T-bpy)](2+). Using spectrophotometry we established the sensor's ability to detect and measure soluble NO in a concentration-dependent manner in cell-free media. Endothelial cells cultured with acetylcholine or hydrogen peroxide to induce endogenous NO production showed modest increases of 7.3 +/- 7.1% and 36.3 +/- 25.0% respectively in fluorescence signal from baseline state, while addition of exogenous NO increased their fluorescence by 5.2-fold. The changes in fluorescence signal were proportionate and comparable against conventional NO assays. Rabbit blood samples immediately exposed to [Ru(bpy)(2)(dabpy)](2+) displayed 8-fold higher mean fluorescence, relative to blood without sensor. Approximately 14% of the observed signal was NO/NO adduct-specific. Optimal readings were obtained when sensor was added to freshly collected blood, remaining stable during subsequent freeze-thaw cycles. Clinical studies are now required to test the utility of [Ru(bpy)(2)(dabpy)](2+) as a sensor to detect changes in NO from human blood samples in cardiovascular health and disease.

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