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

Achini Vidanapathirana, Benjamin Pullen, Run Zhang, Myngan Duong, Jarrad Matthew Goyne, Xiaozhou Zhang, Claudine S. Bonder, Andrew D. Abell, Christina Bursill, S.J. Nicholls, Peter Psaltis

Research output: Contribution to journalArticle

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.
LanguageEnglish
Article number(2019) 9:1720
Number of pages16
JournalScientific Reports
Publication statusPublished - 8 Feb 2019

Keywords

  • Nitric oxide
  • Endothelial cell
  • Ruthenium

Cite this

Vidanapathirana, A., Pullen, B., Zhang, R., Duong, M., Goyne, J. M., Zhang, X., ... Psaltis, P. (2019). A Novel Ruthenium-based Molecular Sensor to Detect Endothelial Nitric Oxide. Scientific Reports, [(2019) 9:1720].
Vidanapathirana, Achini ; Pullen, Benjamin ; Zhang, Run ; Duong, Myngan ; Goyne, Jarrad Matthew ; Zhang, Xiaozhou ; Bonder, Claudine S. ; Abell, Andrew D. ; Bursill, Christina ; Nicholls, S.J. ; Psaltis, Peter. / A Novel Ruthenium-based Molecular Sensor to Detect Endothelial Nitric Oxide. In: Scientific Reports. 2019.
@article{a64e79b297de485e8db262ecfc07b172,
title = "A Novel Ruthenium-based Molecular Sensor to Detect Endothelial Nitric Oxide",
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. Thechanges 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.",
keywords = "Nitric oxide, Endothelial cell, Ruthenium",
author = "Achini Vidanapathirana and Benjamin Pullen and Run Zhang and Myngan Duong and Goyne, {Jarrad Matthew} and Xiaozhou Zhang and Bonder, {Claudine S.} and Abell, {Andrew D.} and Christina Bursill and S.J. Nicholls and Peter Psaltis",
year = "2019",
month = "2",
day = "8",
language = "English",
journal = "Scientific Reports",
issn = "2045-2322",
publisher = "Nature Publishing Group",

}

Vidanapathirana, A, Pullen, B, Zhang, R, Duong, M, Goyne, JM, Zhang, X, Bonder, CS, Abell, AD, Bursill, C, Nicholls, SJ & Psaltis, P 2019, 'A Novel Ruthenium-based Molecular Sensor to Detect Endothelial Nitric Oxide', Scientific Reports.

A Novel Ruthenium-based Molecular Sensor to Detect Endothelial Nitric Oxide. / Vidanapathirana, Achini; Pullen, Benjamin; Zhang, Run; Duong, Myngan; Goyne, Jarrad Matthew; Zhang, Xiaozhou; Bonder, Claudine S.; Abell, Andrew D.; Bursill, Christina; Nicholls, S.J.; Psaltis, Peter.

In: Scientific Reports, 08.02.2019.

Research output: Contribution to journalArticle

TY - JOUR

T1 - A Novel Ruthenium-based Molecular Sensor to Detect Endothelial Nitric Oxide

AU - Vidanapathirana, Achini

AU - Pullen, Benjamin

AU - Zhang, Run

AU - Duong, Myngan

AU - Goyne, Jarrad Matthew

AU - Zhang, Xiaozhou

AU - Bonder, Claudine S.

AU - Abell, Andrew D.

AU - Bursill, Christina

AU - Nicholls, S.J.

AU - Psaltis, Peter

PY - 2019/2/8

Y1 - 2019/2/8

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. Thechanges 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. Thechanges 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.

KW - Nitric oxide

KW - Endothelial cell

KW - Ruthenium

M3 - Article

JO - Scientific Reports

T2 - Scientific Reports

JF - Scientific Reports

SN - 2045-2322

M1 - (2019) 9:1720

ER -

Vidanapathirana A, Pullen B, Zhang R, Duong M, Goyne JM, Zhang X et al. A Novel Ruthenium-based Molecular Sensor to Detect Endothelial Nitric Oxide. Scientific Reports. 2019 Feb 8. (2019) 9:1720.