Large stokes shift and near-infrared fluorescent probe for bioimaging and evaluating the HClO in an rheumatoid arthritis mouse model.

It is crucial to identify aberrant HClO levels in living things since they pose a major health risk and are a frequent reactive oxygen species (ROS) in living organisms. In order to detect HClO in various biological systems, we created and synthesized a near-infrared fluorescent probe with an oxime group (-C = N-OH) as a recognition unit. The probe DCMP1 has the advantages of fast response (10 min), near-infrared emission (660 nm), large Stokes shift (170 nm) and high selectivity. This probe DCMP1 not only detects endogenous HClO in living cells, but also enables further fluorescence detection of HClO in living zebrafish. More importantly, it can also be used for fluorescence imaging of HClO in an rheumatoid arthritis mouse model. This fluorescent probe DCMP1 is anticipated to be an effective tool for researching HClO.

Fast and easy detection of hypochlorite by a smartphone-based fluorescent turn-on probe: Applications to water samples, zebrafish and plant imaging.

We have developed a fluorescent probe DBT-Cl ((E)-2-(2-(4-(diphenylamino)benzylidene) hydrazinyl)-N,N,N-trimethyl-2-oxoethan-1-aminium chloride) for ClO- with an aggregation-induced emission (AIE) strategy depending on solvent polarity. DBT-Cl possessed a prominent solvatochromic emission property with intramolecular charge transfer (ICT) from the TPA (triphenylamine) to the amide group, which was studied by spectroscopic analysis and DFT calculations. These unique AIE properties of DBT-Cl led to the recognition of ClO- with high fluorescent selectivity. DBT-Cl quickly detected ClO- in less than 1 sec with a fluorescent color change from green to cyan. DBT-Cl had a low detection limit of 9.67 µM to ClO-. Detection mechanism of DBT-Cl toward ClO- was illustrated to be oxidative cleavage of DBT-Cl by 1H NMR titrations, ESI-mass, and DFT calculations. We established the viability for dependable detection of ClO- in actual water samples, as well as zebrafish and plant imaging. In particular, DBT-Cl was capable of easily monitoring ClO- through a smartphone application. Therefore, DBT-Cl assured a promising approach for a fast-responsive and multi-applicable ClO- probe in environmental and living organism systems.

Dual-Emissive Detection of ATP and Hypochlorite Ions for Monitoring Inflammation-Driven Liver Injury In Vitro and In Vivo.

Reactive oxygen species play a pivotal role in liver disease, contributing to severe liver damage and chronic inflammation. In liver injury driven by inflammation, adenosine-5'-triphosphate (ATP) and hypochlorite ion (ClO-) emerge as novel biomarkers, reflecting mitochondrial dysfunction and amplified oxidative stress, respectively. However, the dynamic fluctuations of ATP and ClO- in hepatocytes and mouse livers remain unclear, and multidetection techniques for these biomarkers are yet to be developed. This study presents RATP-NClO, a dual-channel fluorescent bioprobe capable of synchronously detecting ATP and ClO- ions. RATP-NClO exhibits excellent selectivity and sensitivity for ATP and ClO- ions, demonstrating a dual-channel fluorescence response in a murine hepatocyte cell line. Upon intravenous administration, RATP-NClO reveals synchronized ATP depletion and ClO- amplification in the livers of mice with experimental metabolic dysfunction-associated steatohepatitis (MASH). Through a comprehensive analysis of the principal mechanism of the developed bioprobe and the verification of its reliable detection ability in both in vitro and in vivo settings, we propose it as a unique tool for monitoring changes in intracellular ATP and ClO- level. These findings underscore its potential for practical image-based monitoring and functional phenotyping of MASH pathogenesis.

An Oxidative Cleavage-Based Cruciform DNA Nanostructure for In Vivo Hypochlorous Acid Visualization to Monitor Intestinal Inflammation.

Ulcerative colitis is a persistent inflammatory bowel disease characterized by inflammation and ulceration in the colon and gastrointestinal tract. It was indicated that the generation of hypochlorous acid (HClO) through the enzymatic activity of myeloperoxidase is significantly linked to ulcerative colitis. In this study, by assembling two hairpins (Hpa and Hpb) onto a quadrivalent cruciform DNA nanostructure, a novel HClO-activatable fluorescent probe was developed based on DNA nanomaterials (denoted MHDNA), which is sensitive, economic, simple, and stable. In the presence of HClO, the Trigger (T) was liberated from the MHDNA probe through a hydrolysis reaction between HClO and phosphorothioate (PS), which is modified on the MHDNA probe and has proved to exhibit particular susceptibility to the HClO. The liberated T subsequently initiated the opening of Hpa and Hpb to facilitate the catalyzed hairpin assembly (CHA) reaction, resulting in the changes of fluorescence and releasing T for recycled signal amplification to achieve sensitive detection of HClO (with a limit of detection 9.83 nM). Additionally, the MHDNA-based spatial-confinement effect shortens the physical distance between Hpa and Hpb and yields a high local concentration of the two reactive hairpins, achieving more rapid reaction kinetics in comparison to conventional CHA methods. Inspirationally, the MHDNA probe was effectively utilized for imaging HClO in ulcerative colitis mice, yielding valuable diagnostic insights for ulcerative colitis.

A new FRET-based fluorescent probe: Colorimetric and ratiometric detection of hypochlorite and anti-counterfeiting applications.

Hypochlorite (ClO-), as the main component of widely used disinfectants in daily life, comes into closer contact with the human body, which can lead to a number of diseases. The high-performance method is increasingly needed to detect ClO- in our daily life. In this report, we successfully synthesized a FRET ratiometric fluorescent probe (NDAC) containing benzoxadiazole moieties and coumarin moieties bound via ethylenediamine. As expected, NDAC has excellent selectivity and anti-interference ability toward ClO-, and the ratio of fluorescence intensity (I471 nm/I533 nm) has a very good linear relationship with the concentration of ClO-, with a wide linear range (2.5-1750 µM) and low detection limit (0.887 µM). Furthermore, we have successfully applied it for the quantitative detection of ClO- in water samples in daily life. At the same time, there is a very clear change in the fluorescence color after the reaction of the NDAC with ClO-. The blue/green value (B/G) of this color change also shows a very good linear relationship to ClO- (5.0-1000 µM). Therefore, the NDAC has also been successfully used for test strip detection and quantitative detection of ClO- in actual samples through smartphone-based fluorescence image analysis, and this method can provide faster, more convenient and more accessible detection. In addition, NDAC sensors also have potential applications in the field of information anti-counterfeiting.

Detection and application of hypochlorous acid in both aqueous environments and living organisms.

The scarcity of water resources has raised concerns regarding drinking water safety. Excessive addition of hypochlorous acid (OCl-) as a disinfectant in drinking water can result in severe consequences. Moreover, abnormal levels of OCl- within the human body can lead to various diseases. Employing fluorescence analysis, the design and synthesis of specific fluorescent probes for simultaneous detection of OCl- in water environments and living organisms holds strategic significance in ensuring the safety of drinking water and mitigating potential risks caused by its abnormal concentrations. This article utilizes naphthalimide as a precursor to develop a novel probe enabling highly sensitive detection of OCl- in water environments and at the organelle level within living organisms. This endeavor serves to provide assurance for drinking water safety and offers health alerts.

Taming Janus-Faced Quinoline-Derived Fluorescent Probes for Dual-Channel Distinguishable Visualization of HSO3- and HClO in Dried Foods and Living Cells.

With the booming development of food manufacturing, developing ideal analytical tools to precisely quantify food additives is highly sought after in the food science field. Herein, a new series of quinoline-derived multifunctional fluorescent probes has been synthesized. Bearing double reactive sites, these compounds display fluorescence response toward both bisulfite (HSO3-) and hypochlorous acid (HClO). Among these compact structures, compound ethyl-2-cyano-3-(6-(methylthio)quinolin-2-yl)acrylate (QTE) was screened out. Probe QTE not only shows ratiometric variation toward HSO3- with little cross talk but also performs turn-off signal toward HClO. In addition, probe QTE has been utilized for bioimaging of HClO in living cells. Furthermore, the HSO3- content in dried food samples has been appraised by QTE with satisfactory results. Meanwhile, relying on the apparent chromaticity change, a flexible dark-box device has been elaborated for chromatic analysis, promoting visualization of HSO3- in the field.

Dicyanoisophorone-based near-infrared fluorescent probe with large Stokes shift for the monitoring and bioimaging of hypochlorite.

Hypochlorite (ClO-), as one of reactive oxygen species (ROS), is closely linked to various illnesses and is essential for the proper functioning of immune system. Hence, monitoring and assessing ClO- levels in organisms are extremely important for the clinical diagnosis of ClO--related disorders. In this study, a novel ClO--selective fluorescent probe, DCP-ClO, was synthesized with dicyanoisophorone-xanthene unit as parent fluorophore, which displayed excellent selectivity towards ClO-, near-infrared emission (755 nm), large Stokes shift (100 nm), real-time response to ClO-, high sensitivity (LOD = 3.95 × 10-8 M), and low cytotoxicity. The recognition mechanism of DCP-ClO towards ClO- was confirmed to be a typical ICT process by HPLC-MS, HR-MS, 1H NMR and theoretical calculations. Meanwhile, DCP-ClO demonstrated remarkable efficacy in monitoring ClO- levels in water samples and eye-catching ability in imaging endogenous/exogenous ClO- in living organisms, which verified its potential as a powerful tool for the recognition of ClO- in complex biological systems.

Persistent Luminescence Lifetime-Based Near-Infrared Nanoplatform via Deep Learning for High-Fidelity Biosensing of Hypochlorite.

In light of deep tissue penetration and ultralow background, near-infrared (NIR) persistent luminescence (PersL) bioprobes have become powerful tools for bioapplications. However, the inhomogeneous signal attenuation may significantly limit its application for precise biosensing owing to tissue absorption and scattering. In this work, a PersL lifetime-based nanoplatform via deep learning was proposed for high-fidelity bioimaging and biosensing in vivo. The persistent luminescence imaging network (PLI-Net), which consisted of a 3D-deep convolutional neural network (3D-CNN) and the PersL imaging system, was logically constructed to accurately extract the lifetime feature from the profile of PersL intensity-based decay images. Significantly, the NIR PersL nanomaterials represented by Zn1+xGa2-2xSnxO4: 0.4 % Cr (ZGSO) were precisely adjusted over their lifetime, enabling the PersL lifetime-based imaging with high-contrast signals. Inspired by the adjustable and reliable PersL lifetime imaging of ZGSO NPs, a proof-of-concept PersL nanoplatform was further developed and showed exceptional analytical performance for hypochlorite detection via a luminescence resonance energy transfer process. Remarkably, on the merits of the dependable and anti-interference PersL lifetimes, this PersL lifetime-based nanoprobe provided highly sensitive and accurate imaging of both endogenous and exogenous hypochlorite. This breakthrough opened up a new way for the development of high-fidelity biosensing in complex matrix systems.

A lysosome-targeted fluorescent probe for fluorescence imaging of hypochlorous acid in living cells and in vivo.

Lysosomes, as crucial acidic organelles in cells, play a significant role in cellular functions. The levels and distribution of hypochlorous acid (HOCl) within lysosomes can profoundly impact their biological functionality. Hence, real-time monitoring of the concentration of HOCl in lysosomes holds paramount importance for further understanding various physiological and pathological processes associated with lysosomes. In this study, we developed a bodipy-based fluorescent probe derived from pyridine and phenyl selenide for the specific detection of HOCl in aqueous solutions. Leveraging the probe's sensitive photoinduced electron transfer effect from phenyl selenide to the fluorophore, the probe exhibited satisfactory high sensitivity (with a limit of detection of 5.2 nM and a response time of 15 s) to hypochlorous acid. Further biological experiments confirmed that the introduction of the pyridine moiety enabled the probe molecule to selectively target lysosomes. Moreover, the probe successfully facilitated real-time monitoring of HOCl in cell models stimulated by N-acetylcysteine (NAC) and lipopolysaccharide (LPS), as well as in a normal zebrafish model. This provides a universal method for dynamically sensing HOCl in lysosomes.

Design, synthesis, and biological application of A-D-A-type boranil fluorescent dyes.

For the first time, three acceptor-donor-acceptor (A-D-A)-type boranil fluorescent dyes, CSU-BF-R (R = H, CH3, and OCH3), featuring phenothiazine as the donor, were designed and synthesized. CSU-BF-R exhibited remarkable photophysical characteristics, including large Stokes shifts (>150 nm), high fluorescence quantum yields (up to 40%), long-wavelength emissions, and strong red solid-state fluorescence. Moreover, these CSU-BF-R fluorescent dyes were demonstrated to function as highly selective and sensitive ratiometric fluorescent probes for detecting hypochlorous acid (HClO). The preliminary biological applications of CSU-BF-OCH3 for sensing intracellular HClO in living cells and zebrafish were demonstrated. Therefore, CSU-BF-R possess the potential to further explore the physiological and pathological functions associated with HClO and provide valuable insights into the design of high-performance A-D-A-type fluorescent dyes.

Cyanine based ratio fluorescent probe and its application in hypochlorite detection.

Hypochlorite (ClO-), a weakly acidic reactive oxygen species, plays a crucial role in antibacterial and anti-inflammatory defense mechanisms. However, elevated levels of ClO- or disruptions in endogenous sites can lead to tissue damage and various diseases including cardiovascular disease, neuronal degeneration, and arthritis. To address this, the development of a specific fluorescent probe with a built-in self-calibration ratio mode for the analysis and biological imaging of ClO- is essential. In this study, a cyanine-based fluorescent probe (Cy-H) was designed for ratiometric fluorescent detection of ClO-, utilizing its aggregation behavior as a novel approach in this field. Upon exposure to ClO-, the phenolic hydroxyl group in probe Cy-H was oxidized into benzoquinone, leading to the formation of cyanine products that displayed a strong tendency to aggregate. As a result, the maximum emission peak of the probe shifted from 700 nm to 485 nm. Notably, a linear relationship was observed between the peak intensity ratio (I485/I700) and the concentration of hypochlorite, with a limit of detection (LOD) of 0.49 µM. Furthermore, this probe was successfully employed for imaging analysis of hypochlorite in living cells and zebrafish.

A triphenylamine-based carbohydrazide hydrazone fluorescent probe for selective detection of hypochlorite and sensing acidic gases.

Hypochlorous acid and its hypochlorite are important reactive oxygen species in the body, and are involved in various physiological processes related to immunity; their rapid detection is of great significance. Here, we synthesized a fluorescent probe (TPAS) by condensation of 4-(diphenylamino)benzaldehyde, carbohydrazide, and salicylaldehyde, which can be used for the detection of ClO- in water and sensing of acidic gas in its solid state. The probe showed strong selective recognition of ClO- in acetonitrile and good tolerance to interference ions. There were good linear responses between the intensity of absorbance and fluorescence and the amount of ClO-. The TPAS solid and its paper strips can emit red fluorescence when exposed to volatile acidic vapours. After being treated with NH3, the red fluorescence can be restored to yellow. The response process of TPAS to ClO- and acid gases was characterized using nuclear magnetic resonance, electrospray ionisation mass spectrometry, transmission electron microscopy, and density functional theory calculations. Furthermore, it can be utilized in analyzing ClO- in commercially available bleaching products; the detection results were basically compatible with the labelled values. In addition, the probe is biocompatible and can be applied for imaging ClO- in zebrafish.

De Novo Design of a Highly Stable Ratiometric Probe for Long-Term Continuous Imaging of Endogenous HClO Burst.

Long-term continuous imaging of endogenous HClO burst is of great importance for the elucidation of various physiological or pathological processes. However, most of the currently reported HClO probes have failed to achieve this goal due to their insufficient photobleaching resistance under a laser source. Herein, a highly stable ratiometric probe, HFTC-HClO 1, which is capable of continuously monitoring endogenous HClO burst over a long period of time, has been judiciously developed. Briefly, the de novo development of HFTC-HClO 1 mainly involved three main steps: (1) novel coumarins (HFTC 1-5) were designed and synthesized; (2) the most stable scaffold, HFTC 3, was selected through dye screening and cell imaging validation; and (3) based on HFTC 3, three candidate HClO probes were constructed, and HFTC-HClO 1 was finally selected due to its superior sensing properties toward HClO. Furthermore, HFTC-HClO 1 can quantitatively measure HClO levels in various real samples with excellent recovery (>90.4%), and the use of HFTC-HClO 1-coated test strips for qualitative analysis of HClO in real samples was also achieved. In addition, the application of HFTC-HClO 1 for long-term continuous monitoring of intracellular HClO burst was successfully demonstrated. Significantly, HFTC-HClO 1 was able to visualize HClO generated in the rheumatoid arthritis mouse model.

A facile near-infrared xanthene fluorescence probe for visualizing of hypochlorous acid in vitro and in vivo.

BACKGROUND: Hypochlorous acid (HClO) is an important biomarker for inflammation, cardiovascular disease, and even cancer. It is of great significance to accurately monitor and quantitatively analyze the fluctuations of HClO to better understand their physiological functions. Traditional HClO detection methods such as high-performance liquid chromatography (HPLC), and mass spectrometry are preferred, but are costly and unsuitable in vivo. Near-infrared (NIR) fluorescence imaging has the advantages of high sensitivity, high temporal and spatial resolutions, minimal autofluorescence, and deep tissue penetration, which facilitates its application in biological systems. Therefore, the development of sensitivity and simple NIR fluorescence monitoring HClO methods in vivo and in vitro is essential and desirable. RESULTS: Herein, we present a NIR probe NOF3 by integrating the rhodamine scaffold and HClO-triggered moiety for the real-time detection of HClO in vitro and in vivo. NOF3 reacts with the HClO and releases the NOF-OH fluorophore of emitted signals at 730 nm, which is in the NIR region. The designed probe detected concentrations of HClO ranging from 0 to 17 µM with a low detection limit of 0.146 µM, presenting excellent sensitivity and selectivity toward HClO over other species. NOF3 manifests significantly turn-on NIR fluorescent signals in response to HClO concentration, which makes it favorable for monitoring dynamic HClO distribution in vivo. We exemplify NOF3 for the tracking of endogenously overexpressed HClO distribution in RAW 264.7 cells, and further realize real-time in vivo bioimaging of HClO activity in inflammation mice. SIGNIFICANCE: The facile NIR NOF3 probe was successfully applied to visualize endogenous and exogenous HClO in living cells and mice. This study provides not only an effective tool for spatial and temporal resolution HClO bioimaging in vivo but also possesses great potential for use in future research on HClO-related biology and pathology.

A novel fluorescent probe based on carbazole-thiophene for the recognition of hypochlorite and its applications.

A carbazole thiophene-aldehyde and 4-methylbenzenesulfonhydrazide conjugate CSH was synthesized by introducing 5-thiophene aldehyde at the 3-position of the carbazole group as the precursor and then condensing it with 4-methylbenzenesulfonhydrazide. CSH has high selectivity and sensitivity towards ClO-, which can specifically identify ClO- by UV-Vis and fluorescence spectroscopy. CSH can rapidly respond to ClO- in the physiological pH range through a fluorescence quenching pattern, accompanied by the color of CSH changing markedly from turquoise to yellowish green under the 365 nm UV light. Probe CSH exhibits a quantitative response to ClO- (0-11 µM) with a low detection limit (1.16 × 10-6 M). Cell imaging experiments have shown that CSH can capture fluorescent signals in the cyan and yellow channels of HeLa cells through fluorescence confocal microscopy, and can successfully identify exogenous ClO- in HeLa cells. In addition, probe CSH can also be used to detect ClO- in environmental water samples. These results indicate that CSH has potential application prospects in the environmental analysis and biological aspects.

Fluorescent Biomass-Based Platform for Detection of ClO- in Cells and Water-Soluble Systems.

The generation and presence of excessive hypochlorous acid derivative ionic form (ClO-) could cause various diseases, such as arteriosclerosis, DNA damage, and cardiovascular illness. It is a critical need to develop a highly sensitive sensor for reliable detection of ClO- in cells and water-soluble systems. In this work, a hydroxyl group has been introduced into the compound 2-amino-3-(((E)-4-(2-(2-(2-hydroxyethoxy)ethyl)-1,3-dioxo-2,3-dihydro-1H-benzo[de]isoquinolin-6-yl)benzylidene)amino)maleonitrile (NDC) to increase its solubility in water, at the same time, the hydrazone unit was designed as a specific recognition group for the "off-on" fluorescence probe of ClO-. The probe NDC presents high selectivity, sensitivity, anti-interference, and low detection limit (67 nM) for ClO-. The recognition mechanism that ClO- breaks the C=N bond and forms the fluorescent compound 4-(2-(2-(2-hydroxyethoxy)ethyl)-1,3-dioxo-2,3-dihydro-1H-benzo[de]isoquinolin-6-yl)benzaldehyde (ND-3) has been confirmed by time-of-flight mass spectrometry. The probe NDC presents a good performance in the actual test of water samples and can be designed as the test papers for the quick and convenient detection of ClO- range from 0 to 1 µM. Moreover, the practical application was demonstrated by the successful imaging of endogenous and exogenous ClO- in HeLa cells. Our fluorescent biomass-based platform opens vast possibilities for repeatability, sensitivity, and selectivity detection of ClO- in cells and water-soluble systems.

Visualization of the elevated levels of hypochlorous acid in Alzheimer's disease with a ruthenium(II) complex-based luminescence probe.

Alzheimer's disease (AD) is an age-related neurodegenerative disorder that devastatingly affects people's lives. Accumulating evidence indicates that the pathological progression of AD is inseparably connected with hypochlorous acid (HClO). However, further exploring the biological function remains an open challenging due to a lack of effective tools to image HClO in AD brains. To this end, a ruthenium(II) luminescence probe, Ru-HClO, is developed for quantitative detection and visualization of HClO in nerve cells and AD brains. Ru-HClO shows quenched luminescence due to the PET process (excited electron transfer from Ru(II) center to diaminomaleonitrile) and the CN bond isomerization in the excited state. The HClO-triggered specific cleavage reaction with Ru-HClO cleaves the CN bond to form highly luminescent Ru-COOH. Ru-HClO shows rapid response speed, high sensitivity and selectivity, excellent biocompatibility, which makes the probe to be applied to semi-quantitative analysis of HClO in nerve cells and high-throughput screening of anti-AD drugs in the AD cell model. Moreover, using Ru-HClO as a probe, present work further validated that the elevated levels of HClO secretion were accompanied by the AD progressed. These findings may provide valuable results for figuring out the biological roles that HClO played in AD but also for accelerating anti-AD therapeutic discovery.

A turn-on fluorescence strategy for hypochlorous acid detection based on DNAzyme-assisted cyclic signal amplification.

Hypochlorous acid (HClO) is a crucial active oxygen component and one of the innate immunity's barrier substances in the body. Abnormal fluctuations in HClO concentration can lead to increased oxidative stress, cellular dysfunction, and the onset of various diseases. Thus, developing convenient, affordable, efficient, and sensitive methods to monitor HClO concentration in healthcare and pathophysiology research is highly significant. In this study, we developed a novel fluorescence strategy for HClO detection based on nucleic acid oxidative cleavage and Pb2+-dependent DNAzyme. By introducing a phosphorothioate site in the hairpin-structured nucleic acid sequence, the nucleic acid probe specifically recognized HClO and underwent oxidative cleavage. Upon cleavage, the enzyme strand is liberated, forming DNAzyme. This DNAzyme then cleaves the substrate strand, liberating the initially quenched fluorescent dyes and generating a turn-on fluorescent signal. The enzyme strand produced by the oxidative cleavage of HClO can be repeatedly utilized, thus realizing the cyclic signal amplification to reduce background noise. We verified the detection mechanism of this strategy through stepwise fluorescence spectroscopy analysis and electrophoresis. Under optimal experimental conditions, the method achieved a detection limit of 5.38 nM and a linear range of 1 nM-800 nM. This method demonstrated exceptional performance in actual biological sample testing and presented an exciting opportunity for expanded utilization in clinical diagnosis and medical research.

Dental Bleaching with Phthalocyanine Photosensitizers: Effects on Dentin Color and Collagen Content.

With the increasing demand for tooth bleaching in esthetic dentistry, its safety has been the focus of a comprehensive body of literature. In this context, the aim of the present study was to evaluate the application effects of pentalysine ß-carbonylphthalocyanine zinc (ZnPc(Lys)5)-mediated photodynamic therapy in dentin bleaching and its effects on dentin collagen. We first established a new and reproducible tooth staining model using dentin blocks stained by Orange II and then bleached with ZnPc(Lys)5 (25 µM) and hydrogen peroxide (10% or 30%). Data were analyzed with one- and two-way ANOVA and a significance level of p < 0.05. ZnPc(Lys)5 effectively bleached the dentin samples to an extent comparable to hydrogen peroxide at either 10% or 30% concentrations. Further studies on the dentin morphology, chemical element distribution, and protein constituents, using an electron microscope, energy dispersive spectroscopy, X-ray photoelectron spectroscopy, and SDS-PAGE, demonstrated that treatment with the photosensitizer preserved the dentin structure and, at the same time, the major organic component, collagen type I. For comparison, hydrogen peroxide (10% or 30%) treatment significantly degraded the collagen protein. This work indicated that the photosensitizer exerts potent bleaching effects on dentin staining; importantly, does not damage dentin and its collagen content; and opens up a new strategy to further explore various photosensitizers for the bleaching of both tooth enamel and dentin.