Toluidine Blue Research Articles

Histomorphometric Analysis of Osseointegrated Intraosseous Dental Implants Using Undecalcified Specimens: A Scoping Review

Bone histology and histomorphometry are reliable diagnostic tools for the assessment of the bone–implant interface, material safety and biocompatibility, and tissue response. They allow for the qualitative and quantitative analysis of undecalcified bone specimens. This scoping review aims to identify the most common staining techniques, study models for in vivo experiments, and histomorphometric parameters used for quantitative bone evaluation of osseointegrated dental implants in the last decade. The Web of Science, PubMed, and Scopus databases were searched on 1 July 2024 for relevant articles in English, published in the last ten years, and the data were exported to an MS Excel spreadsheet. A total of 115 studies met the eligibility criteria and were included in the present review. The results indicate that the most common study models are dogs, rabbits, and pigs. Some of the most frequently used methods for the assessment of the bone–implant interface are the Toluidine blue, Stevenel’s blue with Van Gieson, and Levai–Laczko stainings. The results from this study demonstrate that the most commonly used histomorphometric parameters in implant dentistry are the bone-to-implant contact (BIC), bone area fraction occupancy (BAFO), bone area (BA), and bone density (BD). This review presents the recent trends in histomorphometric analysis of dental implants and identifies some research gaps that necessitate further research.

Exploring the photosensitizing potential of Nanoliposome Loaded Improved Toluidine Blue O (NLITBO) Against Streptococcus mutans: An in-vitro feasibility study.

Streptococcus mutans is a major contributor to dental caries due to its ability to produce acid and survive in biofilms. Microbial resistance towards common antimicrobial agents like chlorhexidine and triclosan has shifted the research towards antimicrobial Photodynamic therapy (PDT). In this context, Toluidine Blue O (TBO) is being explored for its photosensitizing properties against Streptococcus mutans. There is a huge variation in the effective concentration of TBO among the current studies owing to the differences in source of and delivery system TBO as well as the time, power and energy densities of light. The primary objectives of this study are to encapsulate improved Toluidine Blue O (ITBO) in nanoliposomes (NLITBO), characterize it, and evaluate its antibacterial photosensitizing potential against Streptococcus mutans suspensions in vitro. ITBO was synthesised as per Indian patent (number -543908). NLITBO was prepared using the thin-film hydration method. Dynamic light scattering experiment determined the vesicle size, polydispersity index (PDI), and zeta potential. Surface features were characterized by Scanning and Transmission Electron microscopy. ITBO release from NLITBO was assessed using the extrapolation method. The antibacterial activity of the NLITBO was determined by evaluating the zone of inhibition (ZOI) in the Streptococcus mutans culture and comparing with 2% chlorhexidine gluconate. The minimum inhibitory concentration (MIC) of NLITBO as a photosensitizer with red light (wavelength 650nm, power density 0.1 W/cm2, energy density 9-9.1 J/ cm2, 90seconds time) was evaluated against Streptococcus mutans cells by colorimetric method in 96 well plate. Percentage drug loading, loading efficiency, yield percentage, vesicle size, PDI, Zeta potential of NLTBO was reported as 9.3±0.4%, 84.4±7.6%, 73.5%, 123.52 nm, 0.57, -39.54mV respectively. Clusters of uni-lamellar nanovesicles with smooth non-perforated surfaces were observed in SEM and TEM. The size of the vesicle was within 100 nm. At 24 hours, a cumulative 79.81% of ITBO was released from NLITBO. Mean ZOI and MIC of NLITBO (1 μg /ml) were found to be 0.7±0.2 mm, 0.6μg/ml respectively. We have synthesized and encapsulated improved Toluidine Blue O (ITBO) in nanoliposomes (NLITBO) and thoroughly characterized the formulation. The antibacterial efficacy of NLITBO without light was demonstrated by ZOI which is similar to 2% chlorhexidine gluconate. MIC of NLITBO as a photosensitiser along with the optimal light parameter was also proposed in this study. These findings suggested that NLITBO could serve as an effective alternative to conventional antibacterial treatments in managing Streptococcus mutans rich biofilms. It can have potential pharmaceutical application in oral health care.

Coronal Cementum and Reduced Enamel Epithelium on Occlusal Surface of Impacted Wisdom Tooth in a Human

Background: There is only limited research on the coronal cementum of a tooth, and the mechanisms of its forming process are not well-defined. This report presents a coronal cementum on the occlusal surfaces of enamel in an impacted wisdom tooth in a human, which is not nearly the cervical portion. Materials and Methods: The tooth (Tooth #1) was derived from a 46-year-old female. Histological analysis, including hematoxylin and eosin (HE) and toluidine blue (TB) staining, and Scanning Electron Microscopy and Energy Dispersive X-ray Spectrometer (SEM-EDS) analysis of the extracted tooth were conducted. Radiographic examination showed that Tooth #1 was horizontally impacted in the maxilla and had the apex of a single root placed between the buccal and palatal roots of Tooth #2. Results: Coronal cementum was distributed widely on the enamel, and reduced enamel epithelium was also found with enamel matrix proteins histologically. The formation of acellular cementum was observed to be more predominant than that of the cellular cementum in Tooth #1. SEM showed that the occlusal cementum connected directly with enamel. Calcium mapping revealed an almost similar occlusal cementum and enamel. In addition, the spectrum of elements in coronal cementum resembled the primary cementum according to SEM-EDS. Discussion: Thus, coronal cementogenesis in impacted human teeth might be related to the existence of reduced enamel epithelium.

Fifty shades of green and blue: autopsy findings after administration of xenobiotics.

Unusual findings during an autopsy may come from peculiarities in the position, shape, size, weight, consistency, smell or color of organs. The following study was triggered by an autopsy case in which an unusual blue-green discoloration of organs, which changed during the autopsy, was noticed. A review of the local autopsy database, selected cases including the antemortem clinical documentation and the literature has been performed to clarify the etiology of these conspicuous discolorations and to evaluate their diagnostic value. The study showed that certain xenobiotics may lead to such discoloration. After systemic administration of methylene blue, darkening blue-green discoloration of organs, especially the brain and heart, can be observed. In addition, the systemic administration of toluidine blue also appears to be capable of causing such discoloration. Beyond that, drugs (like Rohypnol®) or other foreign substances (like detergents) containing warning colors, i.e. indigocarmin (E132) or Brilliant Blue FCF (E133) may cause discolorations of the upper gastrointestinal tract or the urinary bladder respectively. A blue-green, possibly darkening discoloration of organs during autopsy may point towards an antemortem administration of certain xenobiotics. The affected organs give an indication of the possible route of application and the type of substance. A differentiated interpretation of the etiology of such conspicuous discolorations at autopsy should only be made considering the (medical) history and, if necessary, complementary (toxicological) examinations.

Facile Preparation of Cross-Linked Moringa oleifera Seed Hulls Powder/Hydroxyapatite Framework Composite for Efficient Removal of Toluidine Blue and Methyl Violet 2B from Aqueous Solution

AbstractIn this study, adsorption of two cationic dyes, Toluidine Blue (TB) and Methyl violet 2B (MV 2B) from an aqueous solution was achieved by using multifunctional composite material. The formulation of the composite (MO@HA) was obtained by using Moringa oleifera seed hull powder, calcium chloride (CaCl2), and ammonium hydrogenophosphate (NH4)2HPO4 salts. Surface morphology, functional groups, specific surface area, and surface charge of the composite were explored using Scanning electron microscopy (SEM), Fourier Transform infrared spectroscopy (FTIR), BET analysis, and point of zero charge (PZC), respectively. The composite material resulted in a structural change in the surface of the adsorbents, increased oxygen vacancies, enhancement of active sites, and a specific surface area of 735.55 m2 g−1. Different adsorption parameters such as pH, contact time, adsorbent dosage, and initial concentration were evaluated. The adsorption study showed that equilibrium was reached after 60 min, and the optimum adsorption pH for both dyes (TB and MV 2B) was 6. Langmuir, Freundlich, Liu, and Temkin were fitted to describe the adsorption isotherm, both TB and MV 2B had best correlation with Liu isotherm. The maximum adsorption capacity of TB and MV 2B were 341.488 and 182.453 mg g−1, respectively. Adsorption-desorption cycling studies on the adsorbent confirmed its regeneration and reusability after 5 cycles. A possible adsorption mechanism involving electrostatic interactions, n-π bonding, and hydrogen bonding was suggested. These findings highlight a new direction in the development of efficient and sustainable adsorbent in environmental remediation, specifically in the removal of dyes from aqueous solution.

Diagnostic Accuracy of Toluidine Blue Electrospun in Detecting Oral Potentially Malignant Disorders.

Oral potentially malignant disorders (OPMDs) with epithelial dysplasia can progress to oral squamous cell carcinoma (OSCC). Toluidine blue (TB) has been used in oral screening with various methods. This study aimed to evaluate the sensitivity and specificity of TB-containing electrospun fibers in detecting OPMDs with epithelial dysplasia compared to histopathology. The concentrations of polyvinylpyrrolidone (PVP) and TB were optimized for fiber diameter and puncture strength using the design of experiments method with a central composite design. These optimized fibers were tested on 36 suspected OPMD lesions in 30 patients, followed by a biopsy for histopathological examination. The quadratic and two-factor interaction models described the relationship between TB and PVP concentrations and fiber characteristics. The TB-containing electrospun fibers had 77.3% sensitivity and 35.7% specificity in detecting OPMDs with epithelial dysplasia. The TB-containing electrospun fibers offer advantages such as easy application, cost-effectiveness, and enhanced sensitivity in identifying OPMDs with epithelial dysplasia. However, their diagnostic accuracy is insufficient as a standalone diagnostic method, and they should be considered an adjunctive tool for detecting dysplastic changes in the oral epithelium. Thailand Clinical Trials Registry: TCTR20240429002.

Comparison and Contrast of Calcite vs. Dolomite after Heat Treatment to Enhance Toluidine Blue Removal from Water

Significant increase in use of color dyes in modern society exerted a great pressure on environmental and water qualities. As such, studies for the removal of color dyes from water have been conducted extensively. In this study, common Earth materials dolomite and calcite were evaluated and contrasted for their removal of toluidine blue (TB), a cationic dye, before and after heat treatment. An increase by a factor of 3 in TB removal capacity from 3.5 to 10 mmol/kg was achieved after dolomite (Dol) was heated to 800 °C (designated as HDol). In contrast, the TB removal capacity increased by more than 100 times from 2 to 220 mmol/kg after calcite (Cal) was heated to 1000 °C (designated as HCal). For Dol and HDol, the TB removal increased as the solution’s pH increased but decreased with increases in the solution’s ionic strength. For Cal and HCal, the influence of the equilibrium solution’s pH and ionic strength on TB removal was negligible. The free energy of TB sorption on Dol, HDol, and Cal were −7 to −15 kJ/mol. The results suggested different removal mechanism for TB by Dol vs. Cal. X-ray diffraction data for Dol showed a slight increase in calcite content after heat treatment. For Cal, CaO was produced after heating, which converted back to calcite after 24 h of mixing with TB solutions. The significant TB removal by HCal could be attributed to its reaction with CaO. Thus, the best solution is to use freshly treated Cal for the removal of TB from solution.

Exploration of the role of Hibiscus esculentus (okra) mucilage for the removal of phenothiazinium dyes from water body: Spectroscopic, thermodynamic and molecular modeling study

Environmental pollution poses a major problem now a day. Several dyes, in the form of industrial waste, pollute water body and may cause adverse effects to human health. In this paper ADME and toxicity of fives Phenothiazinium group of dyes Methylene blue (MB), Azure A (AA), Azure B (AB), Azure C (AC) and Toluidine Blue O (TBO) were predicted using Swiss ADME and Protox II tools. Results showed these dyes may herm for living organism due to their carcinogenic, mutagenic and hepatotoxic properties. Removal efficiency of these dyes using okra plant product were determined using spectroscopic, thermodynamic and molecular modeling study. It was revealed that these dyes adsorb on the surface of okra leaf mostly at pH 7.0 and the adsorption isotherms were found to fit in Langmuir and Freundlich isotherm model, while Temkin model fails to do this. Mucilage present in different parts of okra plant plays a significant role on removal of these dyes and is able to remove near about 71–92 % of dyes from water body by itself. As this process did not fit in any of above said adsorption isotherm model, it may be suggested that some other mechanism may happen. Further studies explore that these dyes bound to the hydrophobic pocket of mucilage with binding affinity in the order of 105 M−1 and the bindings were exothermic in nature with enthalpy change in the range of − 2.94 to − 4.28 kcal/mole. Molecular docking study validate all the experimental results obtained from spectroscopic and thermodynamic study and enlighten the role of structure of dyes on their binding affinity to mucilage. This paper will help to systematically understand the role of okra plant products on removal efficiency of Phenothiazinium group of dyes with their structural variations.

Electroacupuncture alleviates sciatic nerve injury and inhibits autophagy in rats.

Sciatic nerve injury is a common form of peripheral nerve injury (PNI). It has been suggested that electroacupuncture (EA) stimulation at GB30 and ST36 can improve nerve dysfunction post-PNI. Autophagy is an important factor in the regeneration of sciatic nerves and recovery of motor function. Therefore, we investigated the biological effects of EA and examined whether these were mediated by autophagy in sciatic nerve injury. Mechanical clamping of the sciatic nerve in Sprague-Dawley rats was performed to establish an experimental model of sciatic nerve injury. EA stimulation was administered once daily for 15 min for seven consecutive days beginning 1 week after successful modeling. The recovery of sciatic nerve function was examined via the sciatic functional index (SFI) test. Morphometric analysis was conducted by staining nerve samples with toluidine blue. Autophagy-associated protein levels were measured via Western blotting. EA stimulation at GB30 and ST36 significantly increased the number of myelinated fibers, axonal and fiber diameters, and the thickness of the myelin sheath in our rat model of sciatic nerve injury. In addition, EA stimulation greatly facilitated nerve regeneration following sciatic nerve injury. Moreover, sciatic nerve injury-induced autophagy was inhibited by EA stimulation. EA facilitates recovery of injured sciatic nerves and inhibits autophagy in a rat model.

Photobiological response of mast cells to green and red light-emitting diodes (LEDs) in cutaneous burns.

This study assessed the effects of red and green LEDs on mast cells (MCs) in third-degree burns in 75 Wistar rats, divided into control, red LED (RED), and green LED (GREEN) groups. Animals were irradiated daily with RED (630 nm, 300 mW, 0.779 W/cm2, 9 J/cm2, 30 s) and GREEN (520 nm, 180 mW, 0.467 W/cm2, 60 J/cm2, 30 s). Histological sections stained with toluidine blue were analyzed for total and subtype MCs. Standardized MC counting was performed across the viable lesion area, considering lesion margins, through intact connective tissue and the integrity of skin appendages. No statistically significant differences in MCs 2 (with released granules and intact cell border) were found between groups. Irradiated groups showed increased total MCs at 7, 14, and 21 days (p < 0.05), with a decrease in MCs 1 (intact MCs) at all time points compared to control (p < 0.05). Significant changes in MCs 3 (with massive degranulation and partial or complete disintegration of the cell border) degranulation were noted in RED at 7, 14, and 21 days (p < 0.009) and in GREEN at 14 (p < 0.009) and 32 days (p < 0.028). Results suggest red and green LEDs modulate MC recruitment and degranulation in third-degree burns.

Therapeutic Potential of Vitamin B Complex in Peripheral Nerve Injury Recovery: An Experimental Rat Model Study.

Objectives: Vitamin B complexes are frequently used in clinical practice for peripheral nerve trauma. However, there is a lack of scientific data on their effectiveness. This study aims to investigate the impact of the vitamin B complex on nerve recovery in a rat model of peripheral nerve paralysis. Materials and Methods: Sixty male Wistar Albino rats were divided into six groups. Models of nerve injury, including blunt trauma, nerve incision, and autograft, were performed on all rats approximately 1 cm distal to the sciatic notch. B-complex vitamins were injected intraperitoneally at 0.2 mL/day to the treatment groups. The control groups were given 0.2 mL/day saline. After 1 month, the study was terminated, electromyography (EMG) was performed to measure the conduction velocity, and nerve tissue was taken from the repair line. The sciatic function indexes (SFIs) were calculated and analyzed. The histopathological samples were stained with hematoxylin and eosin and Toluidine blue and examined with a light microscope. Pathologically, myelination, fibrosis, edema, and mast cell densities in the nervous tissue were evaluated. Results: The vitamin B treatment groups demonstrated significant improvements in SFI compared to the control groups, indicating functional improvement in nerve damage (p < 0.05). In the nerve graft group, the vitamin B group showed a shorter latency, higher velocity, and larger peak-to-peak compared to the controls (p < 0.05). In the nerve transection group, the vitamin B group had better latency, velocity, and peak-to-peak values than the controls (p < 0.05). In the crush injury group, the vitamin B group exhibited an improved latency, velocity, and peak-to-peak compared to the controls (p < 0.05). Better myelination, less fibrosis, edema, and mast cells were also in the vitamin B group (p < 0.05). Conclusions: Vitamin B treatment significantly improves nerve healing and function in peripheral nerve injuries. It enhances nerve conduction, reduces fibrosis, and promotes myelination, indicating its therapeutic potential in nerve regeneration.

UV-radiation manufacturing of natural macromolecular products salecan and tannic acid-based functional gel material as superadsorbent for toluidine blue remediation

Adsorbent materials constructed from natural macromolecular products are favored because of their wide range of sources, biodegradability, and environmental friendliness. Salecan is a novel extracellular polysaccharide with ideal physicochemical and biological activities. Here, we have designed a polymer gel through UV-initiated polymerization of [2-(Methacryloyloxy) ethyl] dimethyl-(3-sulfopropyl) ammonium hydroxide (SBMA) in the mixture of salecan and tannic acid. Photopatterned polymerization process allowed in situ formation of gel adsorbent in a mild reaction condition with energy-efficient manner. Batch experiments for toluidine blue (TB) adsorption were carried out as a function of initial dye concentration, solution pH, contact time, and gel dosage to examine the adsorption capacity, potential mechanism, and removal efficiency. Adsorption behavior exhibited a pH-dependence pattern, which was closely related to their swelling and morphological properties. Adsorption process was in conformity to pseudo-second-order kinetic and Langmuir isotherm models, unlocking a chemical adsorption behavior and monolayer-type removal. The maximum adsorption was 490.2 mg/g, which could be considered a superiorly competing value. Additionally, the UV-gel still showed desirable recyclability and maintained the adsorption effectiveness over 95 % after five regeneration cycles. This study opened up new prospects in preparing high performance adsorbent for TB decontamination and laid the foundation for polysaccharide-based adsorption material research.

An electrochemical immunosensor for sensitive and rapid detection of cystatin C based on Fe3O4/AuNPs-MWCNTs@PDA nanocomposite

Serum Cystatin C (CysC) is an impressive marker for early diagnosis of renal dysfunction. In this work, we established a novel electrochemical immunosensor based on Fe3O4/AuNPs-MWCNTs@PDA nanocomposite for the detection of CysC. The Fe3O4/AuNPs-MWCNTs@PDA nanozyme complex by polydopamine encapsulation can not only carry massive detection antibodies, but also bind the electroactive substance toluidine blue (TB) through electrostatic adsorption. By immobilizing AuNPs onto the electrode to bind the capture antibody (Ab1), we constructed a sandwich electrochemical immunosensor with low cost, high sensitivity, and repeatability. The detection range is 3.9–125.0 ng/mL with a significant linear relationship between the current peak difference (ip) and logarithm of the CysC concentration. Moreover, the detection limit of the immunosensor is 0.157 ng/mL. We have successfully utilized this novel immunosensor to detect CysC in human serum samples, and these results have implications for its potential use in clinical application.

Organo cationic dyes adsorption on charged nano-clay: Revealing dual mechanisms through GCS modeling and DFT molecular simulations

This study investigates the adsorption mechanisms of organic cationic dyes, specifically malachite green (MG) and toluidine blue (TB), on sodium montmorillonite clay (Na-MMC). Using montmorillonite sourced from the Nador deposit in Northern Morocco, we aim to model cation exchange with the Gouy-Chapman-Stern (GCS) model. The adsorption capacities are found to be 192.5 meq/100 g for TB and 175.8 meq/100 g for MG at room temperature and uncontrolled pH. X-ray diffraction (XRD) analysis was employed to determine the interlayer spacing of the montmorillonite, contributing to the understanding of adsorption mechanisms. GCS modeling provided insight into the adsorption constants for both dyes, alongside parameters related to the solid/solution interface such as the amount adsorbed per layer, ion concentration in the solution, and clay surface potential. These findings confirmed a significant affinity between the dyes and the clay, aligning with the literature. Moreover, Density Functional Theory (DFT) simulations indicated a stronger affinity of toluidine blue for negatively charged surfaces compared to other dyes.

A multifunctional DNA tetrahedron for imaging, gene therapy, and chemotherapy-phototherapy combination: Binding affinity and anticancer activity

Imaging, silencing cancer-related microRNA, and chemotherapy-phototherapy (CTPT) combination therapy are crucial for cancer diagnosis and drug resistance overcoming. In this study, we designed a multifunctional DNA tetrahedron (MB-MUC1-TD) for the targeted delivery of combined daunorubicin (DAU) + toluidine blue O (TBO). The detection limit of miRNA-21 was determined to be 0.91 nM. The intercalation of DAU and TBO into MB-MUC1-TD was proved by spectroscopic and calorimetric methods. The thermodynamic parameters for the interactions of DAU and/or TBO with MB-MUC1-TD confirmed high drug loading. The first addition of TBO in the ternary system achieved a higher loading of both drugs and a more stable complex structure. Deoxyribonuclease I (DNase I) accelerated the release of DAU and/or TBO loaded in MB-MUC1-TD. Confocal laser scanning microscope demonstrated that MB-MUC1-TD exhibited good imaging ability for miRNA-21 to accurately identify cancer cells, and DAU/TBO was predominantly distributed within the nucleus of cancer cells. In vitro cytotoxicity showed better gene therapy efficacy of MB on MCF-7 cells, better biocompatibility of loaded DAU and TBO on LO2 cells, and stronger synergistic cytotoxicity of DAU + TBO on MCF-7/ADR cells. This study may establish a theoretical foundation for co-loading CTPT combination drugs based on multifunctional DNA nanostructures.

Sonodynamic inactivation of gram-negative and gram-positive bacteria in the presence of phenothiazine compounds toluidine blue and azurin A

BackgroundSonodynamic antimicrobial chemotherapy (SACT) is an effective antimicrobial treatment that can avoid the production of drug-resistant bacteria. Design and development of new high-efficiency sonosensitizers play a key role in the practical application of SACT. MethodsThe bacteriostatic effects of two phenothiazine compounds, toluidine blue (TB) and azure A (AA) combined with ultrasonic (US) on Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus) were studied, and the sonodynamic antibacterial activities of TB and AA were compared. The reactive oxygen species (ROS) and the types of ROS produced in the sonodynamic system were detected and the sonodynamic mechanisms of TB and AA were proposed. ResultsThe sonodynamic bacteriostasis mediated by TB and AA increased with the increasing concentration of sonosensitizer, the extension of sonication time and the increase of reaction temperature. The production of ROS was the main reason that TB and AA had excellent sonodynamic antibacterial performance. Singlet oxygen (1O2) and hydroxyl radical (•OH) were the main ROS types in the sonodynamic antibacterial system. The ROS produced by the combined action of AA and US was higher than that of TB. ConclusionBoth TB and AA displayed excellent sonodynamic antibacterial activities. Moreover, AA had a higher sonodynamic activity than TB. The electron donation effect and steric hindrance effect of the methyl group of phenothiazine parent nucleus of TB might be the cause of the decrease of its sonodynamic activity. These results would provide a valuable reference for the further study of phenothiazines sonosensitizers and their clinical application in SACT.

Facile and controllable hybrid-nanoengineering of MWCNTs/Au@ZIF-8 and AuPt@CeO2 based sandwich electrochemical aptasensor for AFB1 determination in foods and herbs

Herein, a sandwich electrochemical sensing strategy for aflatoxin b1 (AFB1) detection based on hybrid-nanoengineering was presented. First, Au nanoparticle was doped into zeolitic imidazolate framework-8 (ZIF-8) to form Au@ZIF-8 by in-situ growth method, followed by multi-walled carbon nanotubes (MWCNTs) addition to synthesize MWCNTs/Au@ZIF-8 via self-assembly. The structural “confinement effect” of ZIF-8 afforded a microenvironment for Au nanoparticles and SMCNTs in a certain spatial region, giving MWCNTs/Au@ZIF-8 excellent electrochemical property as the substrate material. In addition, Au-Pt bimetallic nanoparticle, which exhibited excellent stability and catalytic activity was loaded on the hollow cerium oxide (CeO2) to form AuPt@CeO2 nanoparticle through one-step aqueous phase reduction. Owning to its high surface-to-volume ratio, satisfied electron transfer efficiency and biocompatibility, massive toluidine blue (TB) and AFB1 antibody (Ab) could be modified on the AuPt@CeO2 to form AuPt@CeO2-Ab-TB, which acted as signal tag for the ultrasensitive assay of AFB1. The proposed electrochemical sensing system exhibited wide detection range (2 × 10-5 − 20 ng/mL) and low detection limit (2.13 fg/mL), which has been successfully applied to AFB1 determination in four real samples. The hybrid nanoengineering presented in this work is an active attempt to prepare high-performance substrate material and signal tag, which provides a new insight for the development of highly sensitive and specific electrochemical sensing systems.

Relationship between autophagy and NLRP3 inflammasome during articular cartilage degradation in oestrogen-deficient rats with streptozotocin-induced diabetes

BackgroundEstrogen deficiency and Diabetes mellitus (DM) cause joint tissue deterioration, although the mechanisms are uncertain. This study evaluated the immunoexpression of autophagy and NLRP3-inflammasome markers, in rat articular cartilage with estrogen deficiency and DM. MethodsTwenty rats were sham-operated (SHAM) or ovariectomized (OVX) and equally allocated into four groups: SHAM and OVX groups administered with vehicle solution; SHAM and OVX groups treated with 60mg/kg/body weight of streptozotocin, intraperitoneally, to induce DM (SHAM-DM and OVX-DM groups). After seven weeks, the rats were euthanized, and their joint knees were processed for paraffin embedding. Sections were stained with haematoxylin-eosin, toluidine blue, safranin-O/fast-green or subjected to picrosirius-red-polarisation method; immunohistochemistry to detect beclin-1 and microtubule-associated protein 1B-light chain 3 (autophagy markers), NLRP3 and interleukin-1β (IL-1β) (inflammasome activation markers), along with matrix metalloproteinase-9 (MMP-9), Nuclear factor-kappa B (NFκB), and Vascular endothelial growth factor A (VEGF-A) were performed. ResultsDeterioration of articular cartilage and subchondral bone were greater in SHAM-DM and OVX-DM groups. Higher percentages of immunolabeled chondrocytes to NLRP3, IL-1β, MMP-9, NFκB, and VEGF-A, as well as lower percentages of chondrocytes immunolabeled to autophagy markers, were noticed in estrogen-deficient and diabetic groups. These differences were greater in the OVX-DM group. Percentages of immunolabeled chondrocytes showed negative correlation between autophagy markers v.s IL-1β, NLRP-3, MMP-9, NFκB, and VEGF-A, along with positive correlation between VEGF-A vs. MMP-9, NFκB, IL-1β, and NLRP3, and MMP-9 vs. NFκB. ConclusionsIn conclusion, autophagy reduction and NLRP3 inflammasome activation in chondrocytes may be implicated in articular cartilage degradation, under estrogen-deficient and DM conditions. Moreover, the combination of estrogen deficiency and DM may potentiate those effects.

B10 cells regulate macrophage polarization to alleviate inflammation and bone loss in periodontitis.

The polarization of macrophages into an anti-inflammatory phenotype is crucial for resolving periodontal inflammation. It has been reported that B10 cells can regulate the immune response of macrophages during inflammation and are also able to regulate inflammation in periodontitis. However, whether B10 cells' regulation function in periodontitis is related to macrophage polarization remains unclear. This study aims to investigate whether B10 cells can regulate macrophage polarization in periodontitis. Macrophages were cocultured with B10 cells in vitro for 5days. After coculture, macrophages were obtained for analysis directly or followed by stimulation with Pg-LPS/IFN-γ or IL-4/IL-13. Flow cytometry and/or reverse transcriptase-polymerase chain reaction (RT-PCR) were employed to detect the expression of IL-1β, iNOS, TNF-α, CD206, and ARG-1 in macrophages. B10 cells were transferred on the 5th day after ligation in wild or macrophage-depletion mice. Toluidine blue and TRAP staining were used to evaluate alveolar bone resorption and osteoclast activation. Immunohistochemistry was employed to detect the expression of CD68, IL-1β, TNF-α, iNOS, ARG-1, and IL-10. Immunofluorescence was used to detect the expression of CD68+CD86+M1 macrophages and CD68+CD206+M2 macrophages. In vitro, B10 cells inhibit the expression of IL-1β, iNOS, and TNF-α in macrophages while increasing the expression of CD206 and ARG-1. In experimental periodontitis, B10 cells inhibit the polarization of CD68+CD86+M1 macrophages and iNOS expression but enhance the polarization of CD68+CD206+M2 macrophages and ARG-1 expression. Importantly, the depletion of macrophages partially weakened the regulation function of B10 cells in periodontitis. B10 cells promote M2 macrophage polarization, inhibit M1 macrophage polarization in periodontitis, and alleviate periodontitis partially by regulating macrophage polarization.

Facile coupling of plasmonic Au-NPs on ZnS NFs as a robust SERS substrate for toluidine blue detection and degradation

BackgroundThe robustness and sensitivity of the surface-enhanced Raman spectroscopy (SERS) technique heavily relies on the development of SERS active materials. A hybrid of semiconductor and plasmonic metals is highly effective as a SERS substrate, which enables the trace level detection of various organic pollutants. ResultsThis approach demonstrates the photodeposition of plasmonic gold nanoparticles (Au-NPs) on the surface of semiconductor-zinc sulfide nanoflowers (ZnS NFs), grown via the hydrothermal route. The synergistic contribution of the charge-transfer phenomenon and localized surface plasmon resonance of the Au-NPs/ZnS NFs makes it an ideal SERS substrate for the detection of organic pollutants, toluidine blue (TB). The proposed material has a high SERS enhancement factor (109), low limit of detection (10−11 M), good reproducibility, selectivity and strong anti-interference ability. Furthermore, the practicability of the Au-NPs/ZnS NFs is explored in real-time water samples, which are obtained with the satisfactory recovery rates. Additionally, the UVC light illumination on the Au-NPs/ZnS NFs has efficiently degraded TB within a time period of 150 min. Significance and noveltyThese finding demonstrate the significance of the proposed Au-NPs/ZnS NFs for SERS based detection and degradation of organic pollutants in real-time samples, highlighting their potential in monitoring and treating water pollutants in wastewater.