Imaging Of Hypochlorous Acid Research Articles

Assessing Early Atherosclerosis by Detecting and Imaging of Hypochlorous Acid and Phosphorylation Using Fluorescence Nanoprobe.

The assessment of early atherosclerosis (AS) is of great significance for the early diagnosis and mechanism research. Herein, a novel nanoprobe PCN@FL is developed to realize the simultaneous detection and imaging of phosphorylation and hypochlorous acid (HClO). The selective recognition of HClO is achieved through the specific interaction between DMTC and HClO, while the levels of phosphorylation are detected via the specific interaction between Zr (IV) and phosphates. The nanoprobe can be utilized to monitor the fluctuations in HClO and phosphate in early atherosclerosis. It is observed that the levels of HClO and phosphate in the serum of early AS mice are higher than those of the normal mice. Ultimately, the levels of hypochlorous acid and phosphorylation in the inner wall of aortic vessels are imaged by two-photon microscope. The results show that the levels of HClO and phosphorylation in the early atherosclerotic mice are significantly higher than those of in normal mice. The nanoprobe provides a suitable fluorescent tool for simultaneous detection and imaging of HClO and phosphorylation, which holds promise for early atherosclerotic disease assessment.

Rational construction of a fluorescent sensor for simultaneous detection and imaging of hypochlorous acid and peroxynitrite in living cells, tissues and inflammatory rat models

Modern medical research indicates that hypochlorous acid (HClO) and peroxynitrite (ONOO–) are important biomarkers of oxidative stress. However, the up- or down-regulation of HClO or ONOO– has been closely associated with the occurrence and development of various diseases. In order to investigate the intrinsic entanglement relationship between HClO and ONOO– and their relationship with the occurrence and development of inflammation, it is very valuable to develop fluorescent sensors that are capable of displaying different signals towards HClO, ONOO– and HClO/ONOO–. In this work, we rationally design and construct a novel robust small organic molecule fluorescent sensor (RhNp-ClO-ONOO) towards ONOO–, HClO and HClO/ONOO– with green, red, and green–red three different fluorescent signal outputs, respectively. RhNp-ClO-ONOO has fast responsive time for HClO (∼60 s) and ONOO– (∼20 s). Also it has markedly low detection limits for HClO (∼25.3 nM) and ONOO– (12.4 nM) respectively. In addition, RhNp-ClO-ONOO could be further shown to detect endogenous HClO/ONOO– in living cells, inflammatory tissues and rat model successfully. Therefore, this novel fluorescent sensor with double responsive moiety can offer a powerful tool for studying the role of HClO and ONOO– and the occurrence and development of inflammatory diseases.

A New Phenylazo-Based Fluorescent Probe for Sensitive Detection of Hypochlorous Acid in Aqueous Solution.

In this paper, we designed and synthesized a novel phenylazo-based fluorescent probe (RHN) for the sensing and imaging of hypochlorous acid (HClO) in mitochondria in living cells. In this process, HClO promoted the oxidation of the phenylazo group to generate a free Rhodol fluorophore moiety, which in turn restored strong fluorescence and realized the detection of HClO. As expected, RHN exhibited high selectivity, high sensitivity and rapid response, with detection limits as low as 22 nM (1.155 ng/mL). Importantly, the results of the cell imaging experiments indicated that RHN has the ability to image and sense HClO in mitochondria, which is of great significance for exploration of the specific role of HClO in both the immune system and diseases.

Imaging of hypochlorous acid in mitochondria using an asymmetric near-infrared fluorescent probe with large Stokes shift.

Small-molecule near-infrared (NIR) imaging facilitates deep tissue penetration, low autofluorescence, non-invasive visualization, and a relatively simple operation. As such it has emerged as a popular technique for tracking biological species and events. However, the small Stokes shift of most NIR dyes often results in a low signal-to-noise ratio and self-quenching due to crosstalk between the excitation and emission spectra. With this research, we developed a NIR-based fluorescent probe WD-HOCl for hypochlorous acid (HOCl) detection using the NIR dye TJ730 as the fluorophore, which exhibits a large Stokes shift of 156 nm, with no crosstalk between the excitation and emission spectra. It contains acyl hydrazide as the responsive group and a pyridinium cation as the mitochondria-targeting group. The fluorescence intensity of WD-HOCl was enhanced by 30.1-fold after reacting with HOCl. Imaging studies performed using BV-2 cells indicated that WD-HOCl could be used for endogenous HOCl detection and imaging in living cells exposed to glucose and oxygen deprivation/reperfusion. Finally, we demonstrated that inhibiting the expression of NOX2 reduced the HOCl levels and the severity of oxidative stress during stroke in a mouse model.

A novel NIR fluorescent probe with fast response and large stokes shift for the detection and imaging of hypochlorous acid in living cells

Hypochlorous acid (HOCl), one of the physiologically important reactive oxygen species (ROS), is engaged in a diverse variety of biological and pathophysiological functions. Excessive HOCl buildup can cause a variety of illnesses, therefore, the identification of events in real-time of abnormal levels of exogenous and endogenous HOCl in biological settings is extremely important. In the present paper, we reported a new near-infrared emissive fluorescent probe D-HBT-HOCl for the detection and bioimaging of HOCl in live cells. With regard to the construction of D-HBT-HOCl, we combined the dicyanoisophorone motif with a 2-aminobenzenethiol unit, resulting in the near-infrared emission wavelength of 700 nm and an ultra-large Stokes shift of 180 nm, which potentially endowed D-HBT-HOCl with superiority in biological imaging. In addition, the N, N-dimethylthiocarbamate served as an efficient recognition group for HOCl as well as a fluorescence quenching group in the probe. The results of the experiments showed that the newly designed probe exhibited excellent sensing capacity, including outstanding selectivity, a high signal-to-noise ratio, and a rapid response time for the detection of HOCl. What's more, this probe was further effectively leveraged for the imaging of both extrinsic and intrinsic HOCl in live cells, demonstrating its versatility, and possessed the potential to be used to further investigate the physiological and pathological activity of HOCl in biological systems.

UV-enhanced oxidative quenching of PFO–PFPV Pdots for ratiometric quantification and imaging of hypochlorous acid in living cells

UV-enhanced oxidative quenching of PFO–PFPV Pdots for ratiometric quantification and imaging of hypochlorous acid in living cells

A Sensitive and Selective Fluorescent Probe for Real-Time Detection and Imaging of Hypochlorous Acid in Living Cells.

Hypochlorous acid (HClO), a reactive oxygen species, plays an essential role in the processes of physiology and pathology via reacting with most biological molecules. The abnormal level of HClO may cause inflammation, especially arthritis. To further understand its key role in inflammation, in situ detection of HClO is necessary. Herein, a water-soluble small molecule fluorescent probe (HDI-HClO) is employed to monitor and identify trace amounts of HClO in the biological system. In the presence of HClO, the probe releases a hydroxyl group emitting strong fluorescence because of the restoration of the intramolecular charge transfer process. Furthermore, this probe displays a 150-fold fluorescence enhancement accompanied by a large Stokes shift and a lower detection limit (8.3 nM). Moreover, the probe can make a rapid response to HClO within 8 s, which provides the possibility of real-time monitoring of intracellular HClO. Based on the advantages of rapid dynamics, good water solubility, and excellent biocompatibility, this probe could effectively monitor the fluctuations of exogenous and endogenous HClO in living cells. The fluorescence imaging of HDI-HClO indicated that it is an excellent potential approach for comprehending the relationship between inflammation and HClO.

Smart Bimodal Imaging of Hypochlorous Acid In Vivo Using a Heterobimetallic Ruthenium(II)-Gadolinium(III) Complex Probe.

A unique heterobimetallic Ru(II)-Gd(III) complex, Ru-AN-Gd, is reported to serve as an effective probe for bimodal phosphorescence-magnetic resonance (MR) imaging of hypochlorous acid (HClO) in vitro and in vivo. The probe was designed by incorporating a MR contrast agent, Gd-DOTA, into a HClO-responsive bipyridine-Ru(II) complex derivative. The specific reaction between Ru-AN-Gd and HClO triggers the cleavage of an ether bond in the probe molecule, resulting in phosphorescence turn-on and MR turn-off responses to HClO. The integration of MR and phosphorescence detection modes allows the probe to be employed for detecting HClO in a quite wide concentration range (0.6-2000 μM) and for imaging HClO at various resolutions ranging from the subcellular level to the whole body without a depth limit. Its applicability was demonstrated by phosphorescence imaging of lysosomal HClO in live cells, visualization of HClO generation in a mouse arthritis model, and bimodal phosphorescence-MR imaging of HClO in drug-induced acute liver and kidney injury of a mouse. The research achievements suggested the potential of Ru-AN-Gd for diagnosis and treatment monitoring of HClO-related disease.

Imaging of Hypochlorous Acid by Fluorescence and Applications in Biological Systems.

In recent decades, HOCl research has attracted a lot of scientists from around the world. This chemical species is well known as an important player in the biological systems of eukaryotic organisms including humans. In the human body, HOCl is produced by the myeloperoxidase enzyme from superoxide in very low concentrations (20 to 400 μm); this species is secreted by neutrophils and monocytes to help fight pathogens. However, in the condition called "oxidative stress", HOCl has the capability to attack many important biomolecules such as amino acids, proteins, nucleotides, nucleic acids, carbohydrates, and lipids; these reactions could ultimately contribute to a number of diseases such as neurodegenerative diseases (AD, PD, and ALS), cardiovascular diseases, and diabetes. In this review, we discuss recent efforts by scientists to synthesize various fluorophores which are attached to receptors to detect HOCl such as: chalcogen-based oxidation, oxidation of 4-methoxyphenol, oxime/imine, lactone ring opening, and hydrazine. These synthetic molecules, involving rational synthetic pathways, allow us to chemoselectively target HOCl and to study the level of HOCl selectivity through emission responses. Virtually all the reports here deal with well-defined and small synthetic molecular systems. A large number of published compounds have been reported over the past years; this growing field has given scientists new insights regarding the design of the chemosensors. Reversibility, for example is considered important from the stand point of chemosensor reuse within the biological system; facile regenerability using secondary analytes to obtain the initial probe is a very promising avenue. Another aspect which is also important is the energy of the emission wavelength of the sensor; near-infrared (NIR) emission is favorable to prevent autofluorescence and harmful irradiation of tissue; thus, extended applicability of such sensors can be made to the mouse model or animal model to help image internal organs. In this review, we describe several well-known types of receptors that are covalently attached to the fluorophore to detect HOCl. We also discuss the common fluorophores which are used by chemist to detect HOCl, Apart from the chemical aspects, we also discuss the capabilities of the compounds to detect HOCl in living cells as measured through confocal imaging. The growing insight from HOCl probing suggests that there is still much room for improvement regarding the available molecular designs, knowledge of interplay between analytes, biological applicability, biological targeting, and chemical switching, which can also serve to further sensor and theurapeutic agent development alike.

A fluorinated bihydrazide conjugate for activatable sensing and imaging of hypochlorous acid by 19F NMR/MRI.

Hypochlorous acid (HClO) is one of the most important reactive oxygen species (ROS) and plays a vital role in many physiological and pathological processes. The comprehensive exploration of mechanistic details and the potential clinical translation necessitate the development of reliable probes for prompt and accurate detection of HClO in complex biological environments. Herein we report a fluorinated bihydrazide conjugate as a 19F NMR/MRI probe with a "turn-on" character for the detection of HClO. This probe could selectively respond to HClO, leading to a significant recovery of 19F signals for 19F NMR/MRI. Activatable sensing and imaging of HClO were achieved with SMMC-7721 cells and nude mice, which demonstrates that this small molecular conjugate could serve as a selective probe for real-time sensing and imaging of HClO in biological systems.

Responsive Upconversion Nanoprobe for Background-Free Hypochlorous Acid Detection and Bioimaging.

Responsive nanoprobes play an important role in bioassay and bioimaging, early diagnosis of diseases and treatment monitoring. Herein, a upconversional nanoparticle (UCNP)-based nanoprobe, Ru@UCNPs, for specific sensing and imaging of hypochlorous acid (HOCl) is reported. This Ru@UCNP nanoprobe consists of two functional components,, i.e., NaYF4 :Yb, Tm UCNPs that can convert near infrared light-to-visible light as the energy donor, and a HOCl-responsive ruthenium(II) complex [Ru(bpy)2 (DNCH-bpy)](PF6 )2 (Ru-DNPH) as the energy acceptor and also the upconversion luminescence (UCL) quencher. Within this luminescence resonance energy transfer nanoprobe system, the UCL OFF-ON emission is triggered specifically by HOCl. This triggering reaction enables the detection of HOCl in aqueous solution and biological systems. As an example of applications, the Ru@UCNPs nanoprobe is loaded onto test papers for semiquantitative HOCl detection without any interference from the background fluorescence. The application of Ru@UCNPs for background-free detection and visualization of HOCl in cells and mice is successfully demonstrated. This research has thus shown that Ru@UCNPs is a selective HOCl-responsive nanoprobe, providing a new way to detect HOCl and a new strategy to develop novel nanoprobes for in situ detection of various biomarkers in cells and early disgnosis of animal diseases.

N-Heterocyclic Carbene Boranes as Reactive Oxygen Species-Responsive Materials: Application to the Two-Photon Imaging of Hypochlorous Acid in Living Cells and Tissues.

N-Heterocyclic carbene (NHC) boranes undergo oxidative hydrolysis to give imidazolium salts with excellent kinetic selectivity for HOCl over other reactive oxygen species (ROS), including peroxides and peroxynitrite. Selectivity for HOCl results from the electrophilic oxidation mechanism of NHC boranes, which stands in contrast to the nucleophilic oxidation mechanism of arylboronic acids with ROS. The change in polarity that accompanies the conversion of NHC boranes to imidazolium salts can control the formation of emissive excimers, forming the basis for the design of the first fluorescence probe for ROS based on the oxidation of B-H bonds. Two-photon microscope (TPM) ratiometric imaging of HOCl in living cells and tissues is demonstrated.

N‐Heterocyclic Carbene Boranes as Reactive Oxygen Species‐Responsive Materials: Application to the Two‐Photon Imaging of Hypochlorous Acid in Living Cells and Tissues

AbstractN‐Heterocyclic carbene (NHC) boranes undergo oxidative hydrolysis to give imidazolium salts with excellent kinetic selectivity for HOCl over other reactive oxygen species (ROS), including peroxides and peroxynitrite. Selectivity for HOCl results from the electrophilic oxidation mechanism of NHC boranes, which stands in contrast to the nucleophilic oxidation mechanism of arylboronic acids with ROS. The change in polarity that accompanies the conversion of NHC boranes to imidazolium salts can control the formation of emissive excimers, forming the basis for the design of the first fluorescence probe for ROS based on the oxidation of B−H bonds. Two‐photon microscope (TPM) ratiometric imaging of HOCl in living cells and tissues is demonstrated.

Enhanced Fluorescence Turn‐on Imaging of Hypochlorous Acid in Living Immune and Cancer Cells

Two closely related phenyl selenyl based boron-dipyrromethene (BODIPY) turn-on fluorescent probes for the detection of hypochlorous acid (HOCl) were synthesized for studies in chemical biology; emission intensity is modulated by a photoinduced electron-transfer (PET) process. Probe 2 intrinsically shows a negligible background signal; however, after reaction with HOCl, chemical oxidation of selenium forecloses the PET process, which evokes a significant increase in fluorescence intensity. The fluorescence intensity of probes 1 and 2 with HOCl involves an ∼18 and ∼50-fold enhancement compared with the respective responses from other reactive oxygen/nitrogen species (ROS/RNS) and low detection limits (30.9 nm for 1 and 4.5 nm for 2). Both probes show a very fast response with HOCl; emission intensity reached a maximum within 1 s. These probes show high selectivity for HOCl, as confirmed by confocal microscopy imaging when testing with RAW264.7 and MCF-7 cells.