Salivary proteomic profile of response to different resistance training protocols: A case report.

Resistance training (RT) with blood flow restriction (BFR) or high intensity (HI) are effective to increase muscle mass. To understand this effect, techniques known as "omics" are used to identify possible biomarkers. This study analyzed the salivary proteomic profile of healthy individuals trained before and after two RT protocols both designed with eight exercises for upper- and lower-limbs, one performed at low percentage of one-maximum repetition (%1RM) with BFR technique, and other at high %1RM (HI) without BRF technique. Four healthy males between 18 and 28 years participated in the study. Stimulated saliva was collected before (BBFR/BHI) and immediately after (ABFR/AHI) the two RT protocols. All protein-related processing was performed using label-free proteomic. The difference in expression between groups was expressed as p < .05 for downregulated proteins and 1-p > .95 for upregulated proteins. There was difference in salivary flow between ABFR and BBFR (p = .005). For HI, 87 proteins were found after the practice and 119 before. Three hemoglobin isoforms were increased in AHI compared with BHI. In the BFR comparison, 105 proteins were identified after (ABFR) and 70 before (BBFR). Among those increased ABFR, we highlight five hemoglobin isoforms and Deleted in malignant brain tumors 1 protein. Between ABFR and AHI, 17 isoforms of histones, Transaldolase, Transketolase, Glyceraldehyde-3-phosphate dehydrogenase, and Antileukoproteinase were decreased ABFR. For HI, there was an increase in proteins related to oxidative stress and metabolism of the musculoskeletal system, compared with BFR. HI seems to induce higher anabolic signaling to muscle mass increase and antiatherosclerotic effects.

Enamel Caries Lesion Depth Obtained by Optical Coherence Tomography and Transverse Microradiography: A Comparative Study.

INTRODUCTION: Visual imaging of subsurface caries lesions is of vital interest in dentistry, which can be obtained by invasive radiography technique as well as by available non-destructive imaging approaches. Thus, as a first step toward the development of a new innovative approach, Spectral-domain optical coherence tomography (SD-OCT) was applied to detect the lesion depth in comparison to the established reference technique (transverse microradiography [TMR]). METHODS: Bovine enamel specimens were demineralized for 5 days, following previous studies. For OCT, the resulting artificial lesions were scanned three-dimensionally (SD-OCT) and semi-automated measured (CarLQuant). For TMR, specimens were sectioned and the lesion depth was manually determined (Inspektor Research System). RESULTS: The range of lesion depth detected with OCT was 24.0-174.0 µm (mouth rinse study), 18.0-178.0 µm (toothpastes study) and with TMR 59.2-198.0 µm (mouth rinse study), 33.2-133.4 µm (toothpastes study). We found a strong correlation between both methods in terms of lesion depth (Spearman rankwith outlierp < 0.001, Rho = 0.75, Spearman rankwithout outlierp = 0.001, Rho = 0.79). The two methods produce similar results (Passing-Bablok regression, 1.16). As deeper is the lesion, the smallest is the difference between both methods as indicated by Bland-Altman-plots. CONCLUSION: Especially in the case of deep lesions, the values obtained by both methods are in agreement, and OCT can potentially substitute TMR to detect and assess lesion depth with the benefit of being non-destructive.

Effectiveness of Fluoride Varnishes for White Spot Lesion Prevention and Remineralization during Orthodontic Treatment: A Randomized Controlled Trial.

INTRODUCTION: This study investigated the effectiveness of titanium tetrafluoride (TiF4) varnish compared to sodium fluoride (NaF) varnish for preventing and remineralizing white spot lesions (WSLs) in individuals undergoing orthodontic treatment. METHODS: This randomized, placebo-controlled, and double-blinded study was conducted with sixty-five adolescents who were selected based on caries activity and then randomized into three parallel groups: G1 (placebo varnish), G2 (5%-NaF varnish) and G3 (4%-TiF4 varnish). Volunteers received varnish application weekly for the first 4 weeks, after 6 (T1) and 12 (T2) months. The measured outcomes included: prevention of new WSLs, and reversal/progression of WSLs, assessed by Nyvad and ICDAS indices, as well as quantitative light-induced fluorescence (QLF). Chi-square, ANOVA, and Kruskall-Wallis tests were applied. The level of significance was set at 0.05, and post hoc Bonferroni test for p values were performed to correct for multiple comparisons. RESULTS: 1,274 teeth were included; 70.5% were Nyvad 0, and 29.5% were Nyvad 1, with no differences between the groups at baseline (T0). Regarding ICDAS, 70.5% were ICDAS 0, 21.6% were ICDAS 1, and 7.9% were ICDAS 2. G1 showed an increasing prevalence of WSLs at T1 and maintained stable at T2; G2 exhibited a decline at T2, while G3 experienced a decrease at T1 and T2 (p < 0.01). Incidence of WSLs at T2 was 10.2% (G1), 5.6% (G2) and 1.4% (G3). The percentage of teeth initially scored as Nyvad 0 that progressed to Nyvad 1 was 13%, 6.8%, and 1% for G1, G2, and G3, respectively. Conversely, the percentage of teeth initially scored as Nyvad 1 that regressed to Nyvad 0 or Nyvad 2 (T0-T2) was 14%, 49.3%, and 74.4%, respectively (p < 0.001). As for ICDAS index, regression was observed in 6.5%, 17.8% and 24%, while progression was observed in 14.9%, 7.7% and 0.9% for G1, G2 and G3, respectively (p < 0.001). Significant differences among the 3 groups for integrated fluorescence loss (mean±SD, G1: -14.28±9.47, G2: -11.10±11.49 and G3: -6.77±11.00) were found at T2 (p < 0.01). CONCLUSION: Both varnishes demonstrated the ability to prevent and remineralize WSLs. However, TiF4 varnish exhibited the most effective control over WSLs during the 12-month orthodontic treatment.

Effect of experimental toothpaste containing hydroxyapatite nanoparticles and propolis, with and without fluoride, on the microcosm biofilm and enamel demineralization.

This study evaluated the antimicrobial and anticaries effects of toothpaste containing hydroxyapatite nanoparticles (nanoHAP - 5 or 10%), xylitol (2 or 3%) and propolis (1 or 2%), associated or not with 1500 ppm fluoride (F). An in vitro model was used with microcosm biofilm produced from a pool of human saliva and McBain saliva (1:50) in the first 8 h of culture on 162 bovine enamel specimens. At the end of the experimental period, analyses of metabolic activity, colony forming units (CFU) and transverse microradiography (TMR) were performed. This study showed a possible decrease in demineralization and increase in remineralization by the commercial toothpaste (1500 ppm F) and for the experimental toothpaste containing the highest concentration of all agents, combined with F. In addition, a reduction in antimicrobial activity possibly caused by propolis and xylitol, mainly in relation to cariogenic bacteria, was observed.

Effect of experimental and commercial artificial saliva formulations on the activity and viability of microcosm biofilm and on enamel demineralization for irradiated patients with head and neck cancer (HNC).

The effect of different artificial saliva formulations on biofilm activity and viability, and on enamel demineralization for head and neck cancer (HNC) patients was evaluated. Irradiated enamel samples were treated (1 min) with BioXtra® or with experimental formulations containing carboxymethylcellulose plus inorganic constituents alone (AS) or containing 0.1 mg mL-1 CaneCPI-5 (AS + Cane), 1.0 mg mL-1 hemoglobin (AS + Hb) or combination of both (AS + Cane + Hb). Phosphate-buffered-saline and chlorhexidine (0.12%) were negative and positive control, respectively. Biofilm was produced from the saliva of five male HNC patients, under 0.2% sucrose exposure for 5 days, and daily treated with the formulations (1 min). No significant effects were observed for the different experimental treatments. BioXtra® significantly reduced lactobacilli, demonstrating antibacterial potential for this group. Chlorhexidine was an effective treatment to significantly reduce all parameters, being an important antimicrobial and anticaries agent. Future in vitro studies must be performed using a new approach for the design of the experimental formulations.

Effect of a Toothpaste Containing Surface Pre-Reacted Glass-Ionomer Filler on the Remineralization of Artificial Carious Enamel Lesions in situ.

This study evaluated the remineralizing effect of a toothpaste containing surface pre-reacted glass-ionomer (S-PRG) filler on demineralized enamel in situ. For this, 180 bovine enamel samples were demineralized by using a microcosm biofilm model for 3 days. Thereafter, the samples were randomly signed to 15 healthy volunteers and to 3 cross-over in situ phases corresponding to the following treatments: (1) toothpaste containing 1,500 ppm F as NaMFP (positive control, Colgate®Cavity Protection), (2) toothpaste containing 5% S-PRG filler (Shofu®), and (3) placebo toothpaste (negative control prepared by Shofu®). Four demineralized enamel blocks were fixed into each palatal appliance per phase. The volunteers wore the appliances for 5 days and were trained to brush their teeth 2 times for 2 min a day, while one drop of the toothpaste's slurry (1:3) was dripped on each sample for the same period. The surface hardness and TMR analyses were done and analyzed by ANOVA/Tukey and t test (p < 0.05). S-PRG filler and Colgate® toothpastes were equally able to improve 2-2.5× enamel remineralization by the analysis of % surface hardness recovery. However, S-PRG toothpaste was the only one able to significantly improve ΔΔZ (the integrated mineral loss recovery: 1,489 ± 503 %vol.µm) compared to placebo (1,050 ± 467 %vol.µm), while Colgate® did not differ from placebo. No differences were seen between the groups with respect to ΔLD. S-PRG filler and Colgate® toothpastes show similar potential to remineralize the lesion surface. However, S-PRG toothpaste is better to recover mineral loss at the subsurface area.

Effect of a sugarcane cystatin on the profile and viability of microcosm biofilm and on dentin demineralization.

To analyze the effect of a sugarcane cystatin (CaneCPI-5) on the microbial profile and viability, as well as on the prevention of dentin demineralization using a microcosm biofilm model. Ninety bovine dentine specimens were divided into five experimental groups according with the solution they were treated for 60 s: (1) PBS (negative control), (2) 0.12% chlorhexidine (positive control), (3) Fluoride (500 ppm F, as NaF), (4) 0.025 mg/ml CaneCPI-5, and (5) 0.05 mg/ml CaneCPI-5. Specimens were incubated with inoculum (McBain's saliva plus human saliva) in the first 8 h, and from then on, they were exposed to McBain saliva containing sucrose and daily treated (60 s) with the solutions for 5 days. Resazurin and colony-forming unit counting assays were performed. Dentin demineralization was measured by transverse micro-radiography (TMR). 0.12% chlorhexidine significantly reduced the metabolic activity of the microcosm biofilm in relation to the negative control and treated groups (p < 0.01). CHX and F significantly reduced the counts of total microorganisms, mutans group streptococci, and lactobacilli when compared with the negative control. None of the treatments was able to significantly reduce dentin demineralization in comparison with the negative control. In the model evaluated, CaneCPI-5 neither altered the microcosm biofilm profile and viability nor protected dentin against demineralization.

A sugarcane cystatin (CaneCPI-5) alters microcosm biofilm formation and reduces dental caries.

The antimicrobial and anticaries effects of CaneCPI-5 were evaluated. Ninety bovine enamel samples were treated for 60 s with either phosphate-buffered-saline (PBS), 0.12% chlorhexidine (CHX), 0.05 mg ml-1 CaneCPI-5, 0.1 mg ml-1 CaneCPI-5 or 0.5 mg ml-1 CaneCPI-5. They were incubated with inoculum (human saliva + McBain's saliva) for the first 8 h. From then until the end of the experiment, the enamel was exposed to McBain saliva with sucrose and, once a day, for 5 days, they were treated with the solutions. At the end of the experimental period, resazurin and viable plate count assays were performed. Enamel demineralization was also measured. All concentrations of CaneCPI-5 and CHX significantly reduced the activity of biofilms compared with PBS. For viable plate counts, all treatments similarly reduced the lactobacilli and total streptococci; for the mutans streptococci, 0.05 mg ml-1 CaneCPI-5 performed better than CHX. All CaneCPI-5 concentrations significantly reduced the integrated mineral loss. This study represents the first step regarding the use of CaneCPI-5 within the concept of acquired enamel pellicle and biofilm engineering to prevent dental caries.

The Effect of Solutions Containing Extracts of Vochysia tucanorum Mart., Myrcia bellaCambess., Matricaria chamomilla L. and Malva sylvestris L. on Cariogenic Bacterial Species and Enamel Caries Development.

This study evaluated the effect of experimental solutions containing plant extracts on bacterial species and enamel caries prevention. Microcosm biofilm was produced from human saliva mixed with McBain saliva (0.2% sucrose) on bovine enamel for 5 days (3 days under anaerobiosis and 2 days under aerobiosis) at 37°C. From the 2nd day, the following treatments were applied (1 × 60 s/day): Vochysia tucanorum (10 mg/mL); Myrcia bella (5 mg/mL); Matricaria chamomilla (80 mg/mL); Malva sylvestris, fluoride, and xylitol (Malvatricin Plus®); 0.12% chlorhexidine (CHX, PerioGard®); and PBS (negative control). The medium pH was measured. Quantitative polymerase chain reaction was performed for the detection of Streptococcus mutans and Lactobacillus spp. Enamel demineralization was measured by spectral-domain optical coherence tomography. The data were compared by means of the Kruskal-Wallis/Dunn, two-way ANOVA/Bonferroni, and ANOVA/Tukey tests (p < 0.05). The pH decreased after sucrose exposure; only CHX reestablished pH >5.5 by the last day. CHX also eliminated Lactobacillusspp., but the other treatments did not differ significantly from PBS. Malvatricin Plus® and CHX eliminated S. mutans, but the other treatments did not differ from PBS. Similar results were seen concerning the reduction of lesion depth and reflectivity. The experimental natural-extract solutions were ineffective against cariogenic bacteria and in preventing the development of enamel caries.

In vitro effect of curcumin-mediated antimicrobial photodynamic therapy on fibroblasts: viability and cell signaling for apoptosis.

Although it was demonstrated that curcumin-mediated antimicrobial photodynamic therapy (aPDT) is effective for reducing the viability of microbial cells and the vitality of oral biofilms, the cytotoxicity of this therapeutic approach for host cells has not been yet elucidated. Hence, the aim of this study was to evaluate the cytotoxicity and apoptotic effects of curcumin-mediated aPDT on mouse fibroblasts. Cells were treated with 0.6 or 6 µmol.L-1 curcumin combined with 0.075 or 7.5 J.cm-2 LED at 455 nm. Cytotoxicity was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and crystal violet (CV) assays, while quantitative reverse transcriptase-PCR (qRT-PCR) was used to assess the expression of Bax, Bad, Bcl-2, VDAC-1, cytochrome C, and Fas-L genes for apoptosis. The differences between groups were detected by Kruskal-Wallis and post hoc Dunn's tests for MTT and CV assays and by ANOVA and post hoc Tukey test for qRT-PCR (P < 0.05). The effect of 0.6 µmol.L-1 curcumin plus 0.075 J.cm-2 LED (minimum parameter) did not differ statistically from control group; however, the combination of 0.6 µmol.L-1 curcumin plus 7.5 J.cm-2 LED reduced viable cells in 34%, while the combinations of 6 µmol.L-1 curcumin plus 0.075 and 7.5 J.cm-2 LED reduced viable cells in 47% and 99%, respectively. aPDT increased significantly the relative expression of Bax/Bcl-2, cytochrome C, VDAC-1, and Fas-L genes, without influence on the ratio Bad/Bcl-2. Therefore, curcumin-mediated aPDT activated Bcl-2 apoptosis signaling pathways in mouse fibroblasts regarding present conditions, reducing the viability of cells with the increase of curcumin concentrations and light energies.

Dentifrices or gels containing MMP inhibitors prevent dentine loss: in situ studies.

OBJECTIVES: Evaluate the effect of dentifrices or gels containing MMP inhibitors on dentine loss in situ. MATERIALS AND METHODS: Acrylic palatal appliances containing bovine dentine blocks were divided into two rows, corresponding to the groups erosion (ERO) and erosion associated with abrasion (ERO+ABR). For ERO, the appliances were immersed in a cola drink for 5 min, 4 times/day, while for ERO+ABR, the blocks were brushed for 15 sec with a dentifrice slurry after the second and third erosive challenges. Ten volunteers took part in study 1 (S1), where the dentifrices evaluated contained 1100 ppm fluoride as NaF, 0.61% green tea extract, or 0.012% chlorhexidine digluconate. Thirteen volunteers participated in study 2 (S2), in which the treatment was performed only once (1 min) with gels containing 400 µM EGCG (EGCG400), 0.012% chlorhexidine, 1 mM FeSO4, 1.23% F (NaF), placebo, or received no treatment. Dentine loss was analyzed by profilometry (µm). RESULTS: Regarding S1, ERO+ABR induced significantly higher dentine loss compared with ERO and all dentifrices tested led to significantly lower dentine loss when compared with placebo. For S2, regardless of the conditions or times of evaluation, gels containing EGCG, CHX, or FeSO4 led to significantly less wear compared with the other groups. CONCLUSION: Both dentifrices and gels containing MMP inhibitors significantly reduced dentine loss. CLINICAL RELEVANCE: Dentifrices and gels containing MMP inhibitors are able to increase the protection against dentine wear, although gels have a better effect when compared with fluoride gel, lasting up to 10 days after a single application.

Salivary Hemoglobin Protects against Erosive Tooth Wear in Gastric Reflux Patients.

OBJECTIVE: Saliva is the most important biological factor to protect against erosive tooth wear (ETW). Gastroesophageal reflux disease (GERD) patients have an increased risk of ETW due to the frequent presence of intrinsic acids in the oral cavity. Remarkably, not all GERD patients suffer from ETW, which might be due to differences in the composition of the saliva. METHODS: This study compared the proteomic profile of saliva in patients (1) with GERD and ETW (basic erosive wear examination, BEWE, score ≥9; GE group) and (2) with GERD without ETW (BEWE = 0; GNE group) using shotgun label-free quantitative proteomic analysis nLC-ESI-MS/MS. The ability of hemoglobin (Hb) to protect against initial enamel erosion caused by a daily 10-s immersion of enamel specimens in 0.01 M HCl (pH 2.3) for 3 days was evaluated in vitro for proof of concept. Surface hardness change was used as response variable. RESULTS: The differential expression of Hb subunits was significantly increased in the GNE group versus the GE group, in particular the Hb α-subunit that showed a >22-fold increase. Expressions of serum albumin (4.5-fold) and isoforms of cytoskeletal keratin type II (>3-fold) were also increased in the GNE group. Proteinase inhibitors, such as α1-antitrypsin and α2-macroglobulin, were only identified in the GNE group. In vitro, Hb (1.0 and 4.0 mg/mL) significantly reduced initial enamel erosion compared to a negative control after 3 days. CONCLUSIONS: Our results indicate that many proteins, with special emphasis on Hb, may be involved in the resistance of GERD patients to the occurrence of ETW. These proteins may be candidates for inclusion in dental products to protect against ETW.

Photobiomodulation therapy can change actin filaments of 3T3 mouse fibroblast.

The purpose of this study was to investigate the effects that photobiomodulation therapy might produce in cells, in particular, related to their structure. Thus, this paper presents the results of morphological changes in fibroblasts following low-intensity light illumination. Mouse fibroblasts were grown on glass coverslips on either 4 kPa or 16 kPa gels, to mimic normal tissue conditions. Cells were photo-irradiated with laser light at either 625 nm or 808 nm (total energies ranging from 34 to 47 J). Cells were fixed at 5 min, 1 h, or 24 h after photo-irradiation, stained for both actin filaments and the cell nucleus, and imaged by confocal microscopy. A non-light exposed group was also imaged. A detailed analysis of the images demonstrated that the total polymerized actin and number of actin filaments decrease, while the nucleus area increases in treated cells shortly after photo-irradiation, regardless of substrate and wavelength. This experiment indicated that photobiomodulation therapy could change the morphological properties of cells and affect their cytoskeleton. Further investigations are required to determine the specific mechanisms involved and how this phenomenon is related to the photobiomodulation therapy mechanisms of action.

Root caries lesions inhibition and repair using commercial high-fluoride toothpastes with or without tri-calcium phosphate and conventional toothpastes containing or not 1.5% arginine CaCO3: an in situ investigation.

OBJECTIVES: This double-blind, crossover, and in situ study evaluated the effect of commercial high-F toothpastes with or without functionalized ß-tri-calcium phosphate (fTCP) and conventional-F toothpaste containing or not 1.5% arginine-CaCO3 in preventing and repairing root caries-like lesions. METHODS: Twelve volunteers wore palatal appliances containing sound (S) and pre-demineralized (PD) bovine root dentin blocks in 4 phases of 14 days/each. They used F toothpastes, 3×/day: (i) 1,450 µg F/g (conventional-F); (ii) 1,450 µg F/g + 1.5% arginine-CaCO3 (conventional-F + arginine/CaCO3); (iii) 5,000 µg F/g (high-F); and (iv) 5,000 µg F/g (high-F + fTCP). Sucrose solution (20%) was dropped 8×/day on the S blocks. The demineralization in the S blocks and the remineralization in the PD ones were estimated by % of surface hardness difference (%SH-S) and % of SH recovery (%SHR-PD), respectively. Caries lesion area (ΔS) was determined in the blocks. Statistical analyses were performed by one-way ANOVA/Kruskal-Wallis and multiple comparisons tests (p < 0.05). RESULTS: Both high-F toothpastes presented similar effect to prevent early caries lesions and it was higher than conventional-F + arginine/CaCO3. All treatments resulted in significantly lower ΔS values compared with conventional-F toothpaste (p < 0.0001). High-F + fTCP showed non-significantly higher preventive and reversible effect compared to high-F. The effect of conventional-F + arginine/CaCO3 was similar to high-F and lower than high-F + fTCP in preventing and reverting in-deep caries lesions. CONCLUSIONS: Overall, high-F toothpastes seem to be more effective and interesting feasible strategy for both prevention and remineralization actions in root dentin. CLINICAL RELEVANCE: With the trend of increasing dentin root caries, this study brings new evidences about the preventive and repair effect of high-F toothpastes with or without fTCP in root caries and support the use of arginine in toothpastes.

Effect of commercial herbal toothpastes and mouth rinses on the prevention of enamel demineralization using a microcosm biofilm model.

This work evaluated the effects of commercial toothpastes and mouth rinses containing natural/herbal agents on biofilm viability, extracellular polysaccharide (EPS) production and on enamel demineralization in vitro. Microcosm biofilm was produced on bovine enamel for 5 days and treated daily with: Orgânico natural® (toothpaste/mouth rinse), Boni Natural Menta & Malaleuca® (toothpaste/mouth rinse), Propolis & Myrrh® (toothpaste), Colgate Total 12 Clean Mint® (toothpaste, positive control), Malvatricin® Plus (mouth rinse), PerioGard® (mouth rinse, positive control) or PBS (negative control). Tom's Propolis & Myrrh® and Colgate Total 12® toothpastes and Malvatricin® Plus and PerioGard® mouth rinses significantly reduced biofilm viability (p < 0.05). Only PerioGard® had significant effects on biofilm thickness and EPS. Despite the indication that Tom's Propolis & Myrrh® significantly reduced lesion depth, only Colgate Total 12® significantly reduced mineral loss. Malvatricin® Plus significantly reduced mineral loss and lesion depth, as did PerioGard®. Some herbal products, Malvatricin® Plus and Tom's Propolis & Myrrh®, showed anticaries effects.

Protective Effect of 4% Titanium Tetrafluoride Varnish on Dentin Demineralization Using a Microcosm Biofilm Model.

This study evaluated the effect of titanium tetrafluoride (TiF4) varnish on the development of dentin carious lesions. Bovine root dentin samples were treated for 6 h with: (A) 4% TiF4 varnish (2.45% F); (B) 5.42% sodium fluoride (NaF) varnish (2.45% F); (C) 2% chlorhexidine (CHX) gel - positive control; (D) placebo varnish; or (E) untreated - negative control (n = 4 × biological triplicate, n = 12). Treated dentin samples were exposed to human saliva mixed with McBain saliva (1:50) for the first 8 h in 24-well plates. Thereafter, the medium was removed, and McBain saliva containing 0.2% sucrose was applied for 16 h. From days 2 to 5, McBain saliva with sucrose was replaced daily (37°C, 5% CO2). The demineralization was measured using transverse microradiography, while the effect on biofilm was analyzed using viability, extracellular polysaccharide (EPS), and lactic acid production assays. The data were statistically analyzed (p < 0.05). All treatments (fluorides and CHX) significantly reduced the biofilm viability compared to placebo varnish and negative control. However, none of them was able to reduce the colony-forming unit counting for total microorganism, total streptococci, and Streptococcus mutans. NaF significantly reduced the number of Lactobacillus sp. compared to negative control. No effect was seen on lactic acid production neither on EPS synthesis, except that CHX significantly reduced the amount of insoluble EPS. Both fluorides were able to reduce dentin demineralization compared to placebo varnish and negative control; TiF4 had a better effect in reducing mineral loss and lesion depth than NaF. Therefore, TiF4 varnish has the best protective effect on dentin carious lesion formation using this model.

Analysis of Polymorphisms in Genes Differentially Expressed in the Enamel of Mice with Different Genetic Susceptibilities to Dental Fluorosis.

Genes expressed during amelogenesis are candidates to increase the risk of dental fluorosis (DF). Thus, this study aimed to evaluate the association between polymorphisms in enamel development genes and susceptibility to DF in mice. Mice of both sexes, representing strains 129P3/J (n = 20; resistant to DF) and A/J (n = 20; susceptible to DF), were divided into 2 groups. Each strain received a diet with a low concentration of fluoride (F) and drinking water containing 0 or 50 mg/L of F for 6 weeks. Clinical evaluation and analysis of Vickers enamel microhardness of the incisors were performed. Livers were collected for genomic DNA extraction. Seventeen genetic polymorphisms in Amelx, Ambn, Ambn, Col14a1, Col1a1, Col5a2, Enam, Fam20a, Fam83h, Foxo1, Klk4, Mmp20, Serpinf1, Serpinh1, Smad3, Tuft1, and Wdr72 were genotyped by real-time PCR using Taqman chemistry. Overrepresentation of alleles and genotypes in DF was evaluated using the χ2 test with an alpha of 5%. The clinical aspects of the enamel and the surface enamel microhardness confirmed the DF condition. In the polymorphisms rs29569969, rs13482592, and rs13480057 in Ambn, Col14a1, and Mmp20, respectively, genotype and allele distributions were statistically significantly different between A/J and 129P3/J strains (p < 0.05). In conclusion, polymorphisms in Ambn, Col14a1, and Mmp20 are associated with the susceptibility to DF.

Low-level fluoride exposure reduces glycemia in NOD mice.

Water fluoridation is the most widespread measure to prevent dental caries but its relationship with the development of type-1 diabetes (T1D), which has been increasing by 2-5% worldwide, is not quite well understood. AIM: This study evaluated if fluoride (F) administered in the drinking water can prevent or reduce the development of T1D in non-obese diabetic (NOD) mice, as well as to explore the underlying mechanisms. MATERIALS AND METHODS: Twenty-four weaning NOD mice received water containing 0, 10 or 50 ppm F for 21 days. Plasma glucose and insulin were analyzed. Quantitative proteomic analysis was conducted in the liver and gastrocnemius muscle. RESULTS: Animals treated with 10 ppm F had significantly lower glucose levels than the control group, but there was no significant difference among the groups in relation to insulin. The % of ß-cell function was significantly higher in the 10 ppm F group. Changes in the proteomic profile of muscle and liver were seen among the groups. In the muscle, the 10 ppm F group presented, when compared with control, increased expression of proteins involved in energy metabolism. The 50 ppm F group, compared with control, presented increased expression of proteins related to muscle contraction, differentiation of brown adipose tissue and apoptosis. For the liver, the 10 ppm F group had increase in proteins involved in energy metabolism and protein synthesis, in respect to control. There was also an increase in isoforms of Glutathione S transferase, which was confirmed by Western blotting. In the group treated with 50 ppm F, proteins related to ROS metabolism and energetic metabolism were altered. CONCLUSION: Increased expression of antioxidant proteins by treatment with low F concentration may possibly help to explain protection against the development of T1D, which should be better explored in future mechanistic studies.

Effect of a mouthrinse containing Malva sylvestris on the viability and activity of microcosm biofilm and on enamel demineralization compared to known antimicrobials mouthrinses.

The purpose of this study was to evaluate the antimicrobial (anti-biofilm) and anti-caries (enamel demineralization prevention) effects of Malva sylvestris (Malvatricin® Plus) compared with known antimicrobial mouthrinses. Microcosm biofilm was produced on enamel, using inoculum from pooled human saliva mixed with McBain saliva (0.2% sucrose) for 14 days. The biofilm was treated with mouthrinses for 1 min day-1. Oral-B® Complete, Listerine® Zero and Malvatricin® Plus had the greatest effect on the reduction of biofilm viability (p < 0.0001). On the other hand, lactic acid production was reduced significantly with PerioGard®, Noplak® Max and Listerine® Zero compared with the control (p < 0.0001). No significant differences were found among the mouthrinses with respect to the colony-forming unit counting (total microorganisms, total streptococci, mutans streptococci and lactobacilli) and extracellular polysaccharide production. Enamel demineralization was reduced significantly with PerioGard®, Noplak® Max and Malvatricin® Plus compared with the control (p < 0.0001). Malva sylvestris has a comparable anti-caries effect to chlorhexidine mouthrinses.

Changes in the Proteomic Profile of Acquired Enamel Pellicles as a Function of Their Time of Formation and Hydrochloric Acid Exposure.

OBJECTIVE: Changes in the protein profile of acquired enamel pellicles (AEP) formed in vivo over different time periods were evaluated after the application of hydrochloric acid (HCl). METHODS: Nine subjects were submitted to dental prophylaxis with pumice. After 3 or 120 min, the teeth were isolated with cotton rolls and 50 µL of 0.1 M HCl (pH 1.0), 0.01 M HCl (pH 2.0), or deionized water were applied on the buccal surface of the teeth for 10 s. The AEP was then collected using an electrode filter paper presoaked in 3% citric acid. After protein extraction, the samples were submitted to reverse-phase liquid chromatography coupled to mass spectrometry (nano LC-ESI-MS/MS). Label-free quantification was performed (Protein Lynx Global Service software). RESULTS: A total of 180 proteins were successfully identified in the AEP samples. The number of identified proteins increased with the time of pellicle formation. Only 4 proteins were present in all the groups (isoforms of IgA, serum albumin, and statherin). The greatest number of proteins identified uniquely in one of the groups was obtained for the groups treated with HCl after 2 h of pellicle formation (approx. 50 proteins). CONCLUSION: Proteins resistant to removal by HCl, such as serum albumin and statherin, were identified even in the short-term AEP. In addition, 120-min pellicles present many proteins that are resistant to removal by HCl. This suggests an increase in protection against intrinsic acids with the time of pellicle formation, which should be evaluated in future studies.