Abstract
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.
| Original language | English |
|---|---|
| Article number | 1720 |
| Number of pages | 16 |
| Journal | Scientific Reports |
| Volume | 9 |
| Issue number | 1 |
| DOIs | |
| Publication status | Published - 1 Dec 2019 |
Cite this
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A Novel Ruthenium-based Molecular Sensor to Detect Endothelial Nitric Oxide. / Vidanapathirana, Achini K.; Pullen, Benjamin J.; Zhang, Run; Duong, My Ngan; Goyne, Jarrad M.; Zhang, Xiaozhou; Bonder, Claudine S.; Abell, Andrew D.; Bursill, Christina A.; Nicholls, Stephen J.; Psaltis, Peter J.
In: Scientific Reports, Vol. 9, No. 1, 1720, 01.12.2019.Research output: Contribution to journal › Article › Research › peer-review
TY - JOUR
T1 - A Novel Ruthenium-based Molecular Sensor to Detect Endothelial Nitric Oxide
AU - Vidanapathirana, Achini K.
AU - Pullen, Benjamin J.
AU - Zhang, Run
AU - Duong, My Ngan
AU - Goyne, Jarrad M.
AU - Zhang, Xiaozhou
AU - Bonder, Claudine S.
AU - Abell, Andrew D.
AU - Bursill, Christina A.
AU - Nicholls, Stephen J.
AU - Psaltis, Peter J.
PY - 2019/12/1
Y1 - 2019/12/1
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=85061272687&partnerID=8YFLogxK
U2 - 10.1038/s41598-019-39123-3
DO - 10.1038/s41598-019-39123-3
M3 - Article
VL - 9
JO - Scientific Reports
JF - Scientific Reports
SN - 2045-2322
IS - 1
M1 - 1720
ER -