An investigation of the corrosion behavior of zinc-coated stainless steel orthodontic wires: the effect of physical vapor deposition method.

BACKGROUND: Releasing of metal ions might implicate in allergic reaction as a negative subsequent of the corrosion of Stainless Steel (SS304) orthodontic wires. The aim of this study was to evaluate the corrosion resistance of zinc-coated (Zn-coated) SS orthodontic wires. METHODS: Zinc coating was applied on SS wires by PVD method. Electrochemical impedance spectroscopy (EIS), Potentiodynamic polarization tests and Tafel analysis methods were used to predict the corrosion behavior of Zn-coated and uncoated SS wires in both neutral and acidic environments. RESULTS: The values of Ecorr ,icorr and Rct ,which were the electrochemical corrosion characteristics, reported better corrosion behavior of Zn-coated SS wires against uncoated ones in both artificial saliva and fluoride-containing environments. Experimental results of the Tafel plot analyses were consistent with that of electrochemical impedance spectroscopy analyses for both biological solutions. CONCLUSION: Applying Zn coating on bare SS orthodontic wire by PVD method might increase the corrosion resistance of the underlying stainless-steel substrate.

Process Intensification for Enhanced Fluoride Removal and Recovery as Calcium Fluoride Using a Fluidized Bed Reactor.

This study explored the feasibility of fluoride removal from simulated semiconductor industry wastewater and its recovery as calcium fluoride using fluidized bed crystallization. The continuous reactor showed the best performance (>90% fluoride removal and >95% crystallization efficiency) at a calcium-to-fluoride ratio of 0.6 within the first 40 days of continuous operation. The resulting particle size increased by more than double during this time, along with a 36% increase in the seed bed height, indicating the deposition of CaF2 onto the silica seed. The SEM-EDX analysis showed the size and shape of the crystals formed, along with the presence of a high amount of Ca-F ions. The purity of the CaF2 crystals was determined to be 91.1% though ICP-OES analysis. Following the continuous experiment, different process improvement strategies were explored. The addition of an excess amount of calcium resulted in the removal of an additional 6% of the fluoride; however, compared to this single-stage process, a two-stage approach was found to be a better strategy to achieve a low effluent concentration of fluoride. The fluoride removal reached 94% with this two-stage approach under the optimum conditions of 4 + 1 h HRT combinations and a [Ca2+]/[F-] ratio of 0.55 and 0.7 for the two reactors, respectively. CFD simulation showed the impact of the inlet diameter, bottom-angle shape, and width-to-height ratio of the reactor on the mixing inside the reactor and the possibility of further improvement in the reactor performance by optimizing the FBR configuration.

Efficacy of seventh generation bonding agents as desensitizers in patients with dentin hypersensitivity: a randomized clinical trial.

BACKGROUND: Dentin hypersensitivity (DH) is one of the most challenging and persistent dental complaints characterized by transient, intense pain triggered by various stimuli. It affects a significant portion of the global population, predominantly those aged 20-40. This study aims to evaluate the desensitizing efficacy of seventh-generation dentin bonding agents (Single Bond Universal by 3 M ESPE and Xeno-V + by Dentsply) against a control group using Bifluorid 12 by Voco in mitigating DH within a month of the follow-up period. METHODS: This was a single-center, parallel-group, double-blind, controlled randomized clinical trial conducted at Dow University of Health Sciences, Karachi, Pakistan. A total of 105 patients with DH were allocated into three groups for this study. The patients were divided into three groups (Single Bond Universal by 3 M ESPE and Xeno-V + by Dentsply) and the control group containing fluoride varnish (Bifluorid 12 by Voco). Discomfort Interval Scale scores and Schiff Cold Air Sensitivity Scale scores were recorded at baseline, immediately after the intervention, after 01 weeks, and after 01 month. RESULTS: All the materials demonstrated a statistically significant reduction in discomfort and sensitivity (DIS scores p-value 0.01) immediately after 01 week and over a period of 01 month after treatment compared with the baseline scores before application, with no single material proving superior over the one-month observation period. The study also provided insights into dental hygiene practices, with a significant majority using a toothbrush and sensitivity patterns, with cold stimuli being the most common cause of sensitivity. CONCLUSION: The study demonstrates that Single Bond Universal, Xeno V+, and Bifluorid 12 are equally effective in reducing dentin hypersensitivity, with no distinct superiority observed over a one-month period. The findings highlight the potential of fluoride varnishes as a less technique-sensitive and cost-effective option for treating DH, offering valuable insights for future research and clinical practice. TRIAL REGISTRATION: NCT04225247 ( https://clinicaltrials.gov/study/NCT04225247 ), Date of Registration: 13/01/2020. (Retrospectively registered).

Effect of fluoride or chitosan toothpaste and at-home bleaching in enamel roughness, tooth color, and staining susceptibility.

PURPOSE: To evaluate how fluoride- or chitosan-based toothpaste used during at-home bleaching affects enamel roughness, tooth color, and staining susceptibility. METHODS: Bovine enamel blocks were submitted to a 14-day cycling regime considering a factorial design (bleaching agent x toothpaste, 2 x 3), with n=10: (1) bleaching with 16% carbamide peroxide (CP) or 6% hydrogen peroxide (HP), and (2) daily exposure of a fluoride (1,450 ppm F-NaF) toothpaste (FT), chitosan-based toothpaste (CBT), or distilled water (control). Then, 24 hours after the last day of bleaching procedure the samples were exposed to a coffee solution. Color (ΔEab, ΔE00, L*, a*, b*) and roughness (Ra, µm) analyses were performed to compare the samples initially (baseline), after bleaching, and after coffee staining. The results were evaluated by linear models for repeated measures (L*, a*, b*, and Ra), 2-way ANOVA (ΔEab, ΔE00) and Tukey's test (α= 0.05). RESULTS: After the at-home bleaching procedure (toothpaste vs. time, P< 0.0001), the toothpaste groups presented a statistically lower Ra than the control (CBT 0.05). After coffee exposure, CBT presented lower ΔEab and ΔE00 values in the HP groups (toothpaste, P< 0.0001), and lower b* and a* values in the CP groups (toothpaste vs. time, P= 0.004). CLINICAL SIGNIFICANCE: Fluoride or chitosan delivered by toothpaste can reduce surface alterations of the enamel during at-home bleaching, without affecting bleaching efficacy.

Effect of treatment with phosphate, casein phosphopeptide and fluoride on the remineralization: in vitro study.

This study aimed to evaluate in vitro the effect protocols and anticaries agents containing casein amorphous calcium fluoride phosphopeptide-phosphate (CPP-ACPF, MI Paste Plus), sodium trimetaphosphate (TMP) and fluoride (F), in remineralization of caries lesions. Bovine enamel blocks with initial caries lesions were divided into groups (n = 12): 1) Toothpaste without F-TMP-MI Plus (Placebo); 2) Toothpaste 1100 ppm F (1100F), 3) 1100F + MI Paste Plus (1100F-MI Paste Plus), 4) Toothpaste with 1100F + Neutral gel with 4,500 ppm F + 5%TMP (1100F + Gel TMP) and 5) Toothpaste with 1100F + Neutral gel with 9,000 ppm F (1100F + Gel F). For the 4 and 5 groups the gel was applied only once for 1 minute, initially to the study. For the 3 group, after treatment with 1100F, MI Paste Plus was applied 2x/day for 3 minute. After pH cycling, the percentage of surface hardness recovery (%SHR); integrated loss of subsurface hardness (ΔKHN); profile and depth of the subsuperficial lesion (PLM); concentrations of F, calcium (Ca) and phosphorus (P) in enamel was determined. The data were analyzed by ANOVA (1-criterion) and Student-Newman-Keuls test (p < 0.001). Treatment with 1100F alone led to ~ 28% higher remineralization when compared to treatment with 1100F associated with MI Paste Plus (p < 0.001). The 1100F and 1100F + Gel F groups showed similar values for %SHR (p = 0.150). 1100F + Gel TMP treatment also remineralized the enamel surface by ~ 30% and 20% when compared to the 1100F + Gel F and 1100F groups (p < 0.001). The lower lesion depth (ΔKHN) was observed for the 1100F + Gel TMP group (p < 0.001), where it was 54% and 44% lower in comparison to the 1100F and 1100F + Gel F groups (p < 0.001). Polarized light microscopy photomicrographs showed subsurface lesions in all groups, but these lesions were present to a lower extent in the 1100F + Gel TMP group (p < 0.001). Treatment with 1100F + Gel TMP promoted an increase in the concentration of Ca in the enamel by ~ 57% and ~ 26% when compared to the 1100F and 1100F + MI Paste Plus groups (p < 0.001), respectively. There were no significant differences between the 1100F, 1100F + MI Paste Plus and 1100F + Gel F groups (p > 0.001). Similar values of P in the enamel were observed in the 1100F, 1100F + MI Paste Plus and 1100F + Gel F groups (p > 0.001), except for the 1100F + Gel TMP group, which presented a high concentration (p < 0.001). We conclude that the 1100F+TMP gel treatment/protocol led to a significant increased remineralization when compared to the other treatments/protocols and may be a promising strategy for patients with early caries lesions.

Actinomycetota bioprospecting from ore-forming environments.

Natural products from Actinomycetota have served as inspiration for many clinically relevant therapeutics. Despite early triumphs in natural product discovery, the rate of unearthing new compounds has decreased, necessitating inventive approaches. One promising strategy is to explore environments where survival is challenging. These harsh environments are hypothesized to lead to bacteria developing chemical adaptations (e.g. natural products) to enable their survival. This investigation focuses on ore-forming environments, particularly fluoride mines, which typically have extreme pH, salinity and nutrient scarcity. Herein, we have utilized metagenomics, metabolomics and evolutionary genome mining to dissect the biodiversity and metabolism in these harsh environments. This work has unveiled the promising biosynthetic potential of these bacteria and has demonstrated their ability to produce bioactive secondary metabolites. This research constitutes a pioneering endeavour in bioprospection within fluoride mining regions, providing insights into uncharted microbial ecosystems and their previously unexplored natural products.

Metabolomics reveals high fructose-1,6-bisphosphate from fluoride-resistant Streptococcus mutans.

BACKGROUND: Fluoride-resistant Streptococcus mutans (S. mutans) strains have developed due to the wide use of fluoride in dental caries prevention. However, the metabolomics of fluoride-resistant S. mutans remains unclear. OBJECTIVE: This study aimed to identify metabolites that discriminate fluoride-resistant from wild-type S. mutans. MATERIALS AND METHODS: Cell supernatants from fluoride-resistant and wild-type S. mutans were collected and analyzed by liquid chromatography-mass spectrometry. Principal components analysis and partial least-squares discriminant analysis were performed for the statistical analysis by variable influence on projection (VIP > 2.0) and p value (Mann-Whitney test, p < 0.05). Metabolites were assessed qualitatively using the Human Metabolome Database version 2.0 ( http://www.hmdb.ca ), or Kyoto Encyclopedia of Genes and Genomes ( http://www.kegg.jp ), and Metaboanalyst 6.0 ( https://www.metaboanalyst.ca ). RESULTS: Fourteen metabolites differed significantly between fluoride-resistant and wild-type strains in the early log phase. Among these metabolites, 5 were identified. There were 32 differential metabolites between the two strains in the stationary phase, 13 of which were identified. The pyrimidine metabolism for S. mutans FR was matched with the metabolic pathway. CONCLUSIONS: The fructose-1,6-bisphosphate concentration increased in fluoride-resistant strains under acidic conditions, suggesting enhanced acidogenicity and acid tolerance. This metabolite may be a promising target for elucidating the cariogenic and fluoride resistant mechanisms of S. mutans.

Synthesis of novel magnetic hydroxyapatite-biomass nanocomposite for arsenic and fluoride adsorption.

A magnetic nanocomposite of hydroxyapatite and biomass (HAp-CM) was synthesized through a combined ultrasonic and hydrothermal method, aiming for efficient adsorption of arsenic (As) and fluoride (F-) from drinking water in natural environments. The characterization of HAp-CM was carried out using TG, FTIR, XRD, SEM, SEM-EDS, and TEM techniques, along with the determination of pHpzc charge. FTIR analysis suggested that coordinating links are the main interactions that allow the formation of the nanocomposite. XRD data indicated that the crystalline structure of the constituent materials remained unaffected during the formation of HAp-CM. SEM-EDS analysis revelated a Ca/P molar ratio of 1.78. Adsorption assays conducted in batches demonstrated that As and F- followed a PSO kinetic model. Furthermore, As adsorption fitting well to the Langmuir model, while F- adsorption could be explained by both Langmuir and Freundlich models. The maximum adsorption capacity of HAp-CM was found to be 5.0 mg g-1 for As and 10.2 mg g-1 for F-. The influence of sorbent dosage, pH, and the presence of coexisting species on adsorption capacity was explored. The pH significantly affected the nanocomposite's efficiency in removing both pollutants. The presence of various coexisting species had different effects on F- removal efficiency, while As adsorption efficiency was generally enhanced, except in the case of PO43-. The competitive adsorption between F- and As on HAp-CM was also examined. The achieved results demonstrate that HAp-CM has great potential for use in a natural environment, particularly in groundwater remediation as a preliminary treatment for water consumption.

Mediation of mitochondrial DNA copy number and oxidative stress in fluoride-related bone mineral density alteration in Chinese farmers.

Excessive fluoride can adversely affect bone mineral density (BMD). Oxidative stress and mitochondrial dysfunction are crucial mechanisms of health damage induced by fluoride. Here, a cross-sectional survey involving 907 Chinese farmers (aged 18-60) was carried out in Tongxu County in 2017, aiming to investigate the significance of mitochondrial DNA copy number (mtDNAcn) and oxidative stress in fluoride-related BMD change. Concentrations of urinary fluoride (UF), serum oxidative stress biomarkers, including total antioxidant capacity (T-AOC), total superoxide dismutase (T-SOD), glutathione peroxidase (GSH-Px), and malondialdehyde (MDA), as well as relative mtDNAcn in peripheral blood were determined. The multivariable linear model and mediation analysis were performed to assess associations between UF, oxidative stress, and relative mtDNAcn with BMD. Results showed that GSH-Px levels increased by 6.98 U/mL [95% confidence interval (CI) 3.41-10.56)] with each 1.0 mg/L increment of UF. After stratification, the T-AOC, relative mtDNAcn, and BMD decreased by 0.04 mmol/L (-0.08 ~ -0.01), 0.29-unit (-0.55 ~ -0.04), and 0.18-unit (-0.33 ~ -0.03) with every 1.0 mg/L elevation of UF in the excessive fluoride group (EFG, adults with UF > 1.6 mg/L), respectively. Furthermore, T-AOC and relative mtDNAcn were favorably related to the BMD in the EFG (ß = 0.82, 95%CI 0.16-1.48 for T-AOC; ß = 0.11, 95%CI 0.02-0.19 for relative mtDNAcn). Mediation analysis showed that relative mtDNAcn and T-AOC mediated 15.4% and 17.1% of the connection between excessive fluoride and reduced BMD, respectively. Findings suggested that excessive fluoride was related to lower BMD in adults, and the decrement of T-AOC and relative mtDNAcn partially mediate this relationship.

A V-shaped bis-coumarin based fluorescence probe for F- detection in tea infusions and potable water and bioimaging applications in living systems.

Fluorine (F) is a pivotal element in the formation of human dental and skeletal tissues, and the consumption of water and tea constitutes a significant source of fluoride intake. However, prolonged ingestion of water and tea with excessive fluoride content can lead to fluorosis, which poses a serious health hazard. In this manuscript, a novel turn-on fluorescent probe DCF synthesized by bis-coumarin and tert-butyldiphenylsilane (TBDPS) was introduced for detecting F- in potable water and tea infusions. By leveraging the unique chemical affinity between fluoride and silicon, F- triggers the silicon-oxygen bond cleavage in DCF, culminating in a conspicuous emission of yellow fluorescence. Validated through a succession of optical tests, this probe exhibits remarkable advantages in terms of superior selectivity, a low detection limit, a large Stokes shift, and robust interference resistance when detecting inorganic fluoride. Moreover, it can serve as portable test strips for on-site real-time identification and quantitative analysis of F-. Furthermore, the application of DCF for in-situ monitoring and imaging of F- in zebrafish and soybean root tissues proved its significant value for F- detection in both animal and plant systems. This probe potentially functions as an efficient instrument for delving into the toxic mechanisms of fluoride in physiological processes.

Lanthanide Inorganic Nanoparticles Enhance Semiconducting Polymer Nanoparticles Afterglow Luminescence for In Vivo Afterglow/Magnetic Resonance Imaging.

Dual/multimodal imaging strategies are increasingly recognized for their potential to provide comprehensive diagnostic insights in cancer imaging by harnessing complementary data. This study presents an innovative probe that capitalizes on the synergistic benefits of afterglow luminescence and magnetic resonance imaging (MRI), effectively eliminating autofluorescence interference and delivering a superior signal-to-noise ratio. Additionally, it facilitates deep tissue penetration and enables noninvasive imaging. Despite the advantages, only a limited number of probes have demonstrated the capability to simultaneously enhance afterglow luminescence and achieve high-resolution MRI and afterglow imaging. Herein, we introduce a cutting-edge imaging platform based on semiconducting polymer nanoparticles (PFODBT) integrated with NaYF4@NaGdF4 (Y@Gd@PFO-SPNs), which can directly amplify afterglow luminescence and generate MRI and afterglow signals in tumor tissues. The proposed mechanism involves lanthanide nanoparticles producing singlet oxygen (1O2) upon white light irradiation, which subsequently oxidizes PFODBT, thereby intensifying afterglow luminescence. This innovative platform paves the way for the development of high signal-to-background ratio imaging modalities, promising noninvasive diagnostics for cancer.

MiRNA-seq and mRNA-seq revealed the mechanism of fluoride-induced cauda epididymal injury.

The widespread presence of fluoride in water, food, and the environment continues to exacerbate the impact of fluoride on the male reproductive health. However, as a critical component of the male reproductive system, the intrinsic mechanism of fluoride-induced cauda epididymis damage and the role of miRNAs in this process are still unclear. This study established a mouse fluorosis model and employed miRNA and mRNA sequencing; Evans blue staining, Oil Red O staining, TEM, immunofluorescence, western blotting, and other technologies to investigate the mechanism of miRNA in fluoride-induced cauda epididymal damage. The results showed that fluoride exposure increased the fluoride concentration in the hard tissue and cauda epididymis, altered the morphology and ultrastructure of the cauda epididymis, and reduced the motility rate, normal morphology rate, and hypo-osmotic swelling index of the sperm in the cauda epididymis. Furthermore, sequencing results revealed that fluoride exposure resulted in differential expression of 17 miRNAs and 4725 mRNAs, which were primarily enriched in the biological processes of tight junctions, inflammatory response, and lipid metabolism, with miR-742-3p, miR-141-5p, miR-878-3p, and miR-143-5p serving as key regulators. Further verification found that fluoride damaged tight junctions, raised oxidative stress, induced an inflammatory response, increased lipid synthesis, and reduced lipid decomposition and transport in the cauda epididymis. This study provided a theoretical basis for developing miRNA as potential diagnostic markers and therapeutic target drugs for this injury.

The effect of pseudoknot base pairing on cotranscriptional structural switching of the fluoride riboswitch.

A central question in biology is how RNA sequence changes influence dynamic conformational changes during cotranscriptional folding. Here we investigated this question through the study of transcriptional fluoride riboswitches, non-coding RNAs that sense the fluoride anion through the coordinated folding and rearrangement of a pseudoknotted aptamer domain and a downstream intrinsic terminator expression platform. Using a combination of Escherichia coli RNA polymerase in vitro transcription and cellular gene expression assays, we characterized the function of mesophilic and thermophilic fluoride riboswitch variants. We showed that only variants containing the mesophilic pseudoknot function at 37°C. We next systematically varied the pseudoknot sequence and found that a single wobble base pair is critical for function. Characterizing thermophilic variants at 65°C through Thermus aquaticus RNA polymerase in vitro transcription showed the importance of this wobble pair for function even at elevated temperatures. Finally, we performed all-atom molecular dynamics simulations which supported the experimental findings, visualized the RNA structure switching process, and provided insight into the important role of magnesium ions. Together these studies provide deeper insights into the role of riboswitch sequence in influencing folding and function that will be important for understanding of RNA-based gene regulation and for synthetic biology applications.

Comparison of the effects of fluoride varnish containing silver nanoparticles and conventional fluoride varnish on the surface microhardness of tooth enamel.

BACKGROUND: Nano-silver fluoride (NSF) has been introduced to improve enamel lesions. The effective use of varnishes is important in the prevention of dental caries. OBJECTIVES: The study aimed to compare the effect of conventional sodium fluoride varnish with the same varnish containing 1% and 2% silver nanoparticles (AgNP) on the surface microhardness of enamel. MATERIAL AND METHODS: The baseline surface microhardness of 40 premolar teeth was measured using a Vickers microhardness tester. After immersing the samples in a demineralizing agent for 24 h, the microhardness was measured again. In group B, a layer of conventional fluoride varnish was applied to the tooth surfaces using a microbrush with soft bristles, following the manufacturer's instructions. Groups C and D were treated with 1% and 2% NSF varnishes, respectively, while group A received no varnish. Surface microhardness tests were conducted on all specimens, including those previously tested. RESULTS: The microhardness of the enamel surface increased significantly in all 3 test groups compared to the microhardness after demineralization (p < 0.05). CONCLUSIONS: Conventional fluoride varnish and fluoride varnishes containing 1% and 2% AgNP are equally effective in remineralizing initial caries.

Fluoride export is required for the competitive fitness of pathogenic microorganisms in dental biofilm models.

Microorganisms resist fluoride toxicity using fluoride export proteins from one of several different molecular families. Cariogenic species Streptococcus mutans and Candida albicans extrude intracellular fluoride using a CLCF F-/H+ antiporter and FEX fluoride channel, respectively, whereas oral commensal eubacteria, such as Streptococcus gordonii, export fluoride using a Fluc fluoride channel. In this work, we examine how genetic knockout of fluoride export impacts pathogen fitness in single-species and three-species dental biofilm models. For biofilms generated using S. mutans with the genetic knockout of the CLCF transporter, exposure to low fluoride concentrations decreased S. mutans counts, synergistically reduced the populations of C. albicans, increased the relative proportion of oral commensal S. gordonii, and reduced properties associated with biofilm pathogenicity, including acid production and hydroxyapatite dissolution. Biofilms prepared with C. albicans with genetic knockout of the FEX channel also exhibited reduced fitness in the presence of fluoride but to a lesser degree. Imaging studies indicate that S. mutans is highly sensitive to fluoride, with the knockout strain undergoing complete lysis when exposed to low fluoride for a moderate amount of time. Biochemical purification of the S. mutans CLCF transporter and functional reconstitution establishes that the functional protein is a dimer encoded by a single gene. Together, these findings suggest that fluoride export by oral pathogens can be targeted by specific inhibitors to restore biofilm symbiosis in dental biofilms and that S. mutans is especially susceptible to fluoride toxicity. IMPORTANCE: Dental caries is a globally prevalent condition that occurs when pathogenic species, including Streptococcus mutans and Candida albicans, outcompete beneficial species, such as Streptococcus gordonii, in the dental biofilm. Fluoride is routinely used in oral hygiene to prevent dental caries. Fluoride also has antimicrobial properties, although most microbes possess fluoride exporters to resist its toxicity. This work shows that sensitization of cariogenic species S. mutans and C. albicans to fluoride by genetic knockout of fluoride exporters alters the microbial composition and pathogenic properties of dental biofilms. These results suggest that the development of drugs that inhibit fluoride exporters could potentiate the anticaries effect of fluoride in over-the-counter products like toothpaste and mouth rinses. This is a novel strategy to treat dental caries.

Legumain-Triggered Macrocyclization of Radiofluorinated Tracer for Enhanced PET Imaging.

Enhancing the accumulation and retention of small-molecule probes in tumors is an important way to achieve accurate cancer diagnosis and therapy. Enzyme-stimulated macrocyclization of small molecules possesses great potential for enhanced positron emission tomography (PET) imaging of tumors. Herein, we reported an 18F-labeled radiotracer [18F]AlF-RSM for legumain detection in vivo. The tracer was prepared by a one-step aluminum-fluoride-restrained complexing agent ([18F]AlF-RESCA) method with high radiochemical yield (RCY) (88.35 ± 3.93%) and radiochemical purity (RCP) (>95%). More notably, the tracer can be transformed into a hydrophobic macrocyclic molecule under the joint action of legumain and reductant. Simultaneously, the tracer could target legumain-positive tumors and enhance accumulation and retention in tumors, resulting in the amplification of PET imaging signals. The enhancement of radioactivity enables PET imaging of legumain activity with high specificity. We envision that, by combining this highly efficient 18F-labeled strategy with our intramolecular macrocyclization reaction, a range of radiofluorinated tracers can be designed for tumor PET imaging and early cancer diagnosis in the future.

Evaluation of toothpastes for treating root carious lesions - a laboratory-based pilot study.

BACKGROUND: Root caries is preventable and can be arrested at any stage of disease development. The aim of this study was to investigate the potential mineral exchange and fluorapatite formation within artificial root carious lesions (ARCLs) using different toothpastes containing 5,000 ppm F, 1,450 ppm F or bioactive glass (BG) with 540 ppm F. MATERIALS AND METHODS: The crowns of each extracted sound tooth were removed. The remaining roots were divided into four parts (n = 12). Each sample was randomly allocated into one of four groups: Group 1 (Deionised water); Group 2 (BG with 540 ppm F); Group 3 (1,450 ppm F) and Group 4 (5,000 ppm F). ARCLs were developed using demineralisation solution (pH 4.8). The samples were then pH-cycled in 13 days using demineralisation solution (6 h) and remineralisation solution (pH 7) (16 h). Standard tooth brushing was carried out twice a day with the assigned toothpaste. X-ray Microtomography (XMT) was performed for each sample at baseline, following ARCL formation and after 13-day pH-cycling. Scanning Electron Microscope (SEM) and 19F Magic angle spinning nuclear magnetic resonance (19F-MAS-NMR) were also performed. RESULTS: XMT results showed that the highest mineral content increase (mean ± SD) was Group 4 (0.09 ± 0.05), whilst the mineral content decreased in Group 1 (-0.08 ± 0.06) after 13-day pH-cycling, however there was evidence of mineral loss within the subsurface for Groups 1, 3 and 4 (p < 0.05). SEM scans showed that mineral contents within the surface of dentine tubules were high in comparison to the subsurface in all toothpaste groups. There was evidence of dentine tubules being either partially or completely occluded in toothpaste groups. 19F-MAS-NMR showed peaks between - 103 and - 104ppm corresponding to fluorapatite formation in Groups 3 and 4. CONCLUSION: Within the limitation of this laboratory-based study, all toothpastes were potentially effective to increase the mineral density of artificial root caries on the surface, however there was evidence of mineral loss within the subsurface for Groups 1, 3 and 4.

Zinc- and Fluoride-Releasing Bioactive Glass as a Novel Bone Substitute.

Bioglass 45S5, a silica-based glass, has pioneered a new field of biomaterials. Bioglass 45S5 promotes mineralization through calcium ion release and is widely used in the dental field, including toothpaste formulations. However, the use of Bioglass 45S5 for bone grafting is limited owing to the induction of inflammation, as well as reduced degradation and ion release. Phosphate-based glasses exhibit higher solubility and ion release than silica-based glass. Given that these glasses can be synthesized at low temperatures (approximately 1,000°C), they can easily be doped with various metal oxides to confer therapeutic properties. Herein, we fabricated zinc- and fluoride-doped phosphate-based glass (multicomponent phosphate [MP] bioactive glass) and further doped aluminum oxide into the MP glass (4% Al-MP glass) to overcome the striking solubility of phosphate-based glass. Increased amounts of zinc and fluoride ions were detected in water containing the MP glass. Doping of aluminum oxide into the MP glass suppressed the striking dissolution in water, with 4% Al-MP glass exhibiting the highest stability in water. Compared with Bioglass 45S5, 4% Al-MP glass in water had a notably reduced particle size, supporting the abundant ion release of 4% Al-MP glass. Compared with Bioglass 45S5, 4% Al-MP glass enhanced the osteogenesis of mouse bone marrow-derived mesenchymal stem cells. Mouse macrophages cultured with 4% Al-MP glass displayed enhanced induction of anti-inflammatory M2 macrophages and reduced proinflammatory M1 macrophages, indicating M2 polarization. Upon implanting 4% Al-MP glass or Bioglass 45S5 in a mouse calvarial defect, 4% Al-MP glass promoted significant bone regeneration when compared with Bioglass 45S5. Hence, we successfully fabricated zinc- and fluoride-releasing bioactive glasses with improved osteogenic and anti-inflammatory properties, which could serve as a promising biomaterial for bone regeneration.

Measurement and analysis of microbial fluoride resistance in dental biofilm models.

The interactions between communities of microorganisms inhabiting the dental biofilm is a major determinant of oral health. These biofilms are periodically exposed to high concentrations of fluoride, which is present in almost all oral healthcare products. The microbes resist fluoride through the action of membrane export proteins. This chapter describes the culture, growth and harvest conditions of model three-species dental biofilm comprised of cariogenic pathogens Streptococcus mutans and Candida albicans and the commensal bacterium Streptococcus gordonii. In order to examine the role of fluoride export by S. mutans in model biofilms, procedures for generating a strain of S. mutans with a genetic knockout of the fluoride exporter are described. We present a case study examining the effects of this mutant strain on the biofilm mass, acid production and mineral dissolution under exposure to low levels of fluoride. These general approaches can be applied to study the effects of any gene of interest in physiologically realistic multispecies oral biofilms.

Computational approaches to investigate fluoride binding, selectivity and transport across the membrane.

The use of molecular dynamics (MD) simulations to study biomolecular systems has proven reliable in elucidating atomic-level details of structure and function. In this chapter, MD simulations were used to uncover new insights into two phylogenetically unrelated bacterial fluoride (F-) exporters: the CLCF F-/H+ antiporter and the Fluc F- channel. The CLCF antiporter, a member of the broader CLC family, has previously revealed unique stoichiometry, anion-coordinating residues, and the absence of an internal glutamate crucial for proton import in the CLCs. Through MD simulations enhanced with umbrella sampling, we provide insights into the energetics and mechanism of the CLCF transport process, including its selectivity for F- over HF. In contrast, the Fluc F- channel presents a novel architecture as a dual topology dimer, featuring two pores for F- export and a central non-transported sodium ion. Using computational electrophysiology, we simulate the electrochemical gradient necessary for F- export in Fluc and reveal details about the coordination and hydration of both F- and the central sodium ion. The procedures described here delineate the specifics of these advanced techniques and can also be adapted to investigate other membrane protein systems.