Hemoglobin Protects Enamel against Intrinsic Enamel Erosive Demineralization.

INTRODUCTION: This study investigated the changes in the acquired enamel pellicle (AEP) proteome when this integument is formed in vivo after treatment with sugarcane-derived cystatin (CaneCPI-5), hemoglobin (HB), and a statherin-derived peptide (StN15), or their combination and then exposed to an intrinsic acid challenge. The effectiveness of these treatments in preventing intrinsic erosion was also evaluated. METHODS: Ten volunteers, after prophylaxis, in 5 crossover phases, rinsed with the following solutions (10 mL, 1 min): control (deionized water-H2O) - group 1, 0.1 mg/mL CaneCPI-5 - group 2, 1.0 mg/mL HB - group 3, 1.88 × 10-5M StN15 - group 4, or a blend of these - group 5. Following this, AEP formation occurred (2 h) and an enamel biopsy (10 µL, 0.01 m HCl, pH 2.0, 10 s) was conducted on one incisor. The biopsy acid was then analyzed for calcium (Arsenazo method). The vestibular surfaces of the other teeth were treated with the same acid. Acid-resistant proteins in the residual AEP were then collected and analyzed quantitatively via proteomics. RESULTS: Compared to control, treatment with the proteins/peptide, mixed or isolated, markedly enhanced acid-resistant proteins in the AEP. Notable increases occurred in pyruvate kinase PKM (11-fold, CaneCPI-5), immunoglobulins and submaxillary gland androgen-regulated protein 3B (4-fold, StN15), Hb, and lysozyme C (2-fold, StN15). Additionally, a range of proteins not commonly identified in the AEP but known to bind calcium or other proteins were identified in groups treated with the tested proteins/peptide either in isolation or as a mixture. The mean (SD, mM) calcium concentrations released from enamel were 3.67 ± 1.48a, 3.11 ± 0.72a, 1.94 ± 0.57b, 2.37 ± 0.90a, and 2.38 ± 0.45a for groups 1-5, respectively (RM-ANOVA/Tukey, p < 0.05). CONCLUSIONS: Our findings demonstrate that all treatments, whether using a combination of proteins/peptides or in isolation, enhanced acid-resistant proteins in the AEP. However, only HB showed effectiveness in protecting against intrinsic erosive demineralization. These results pave the way for innovative preventive methods against intrinsic erosion, using "acquired pellicle engineering" techniques.

Estimated Dietary Fluoride Intake by 24-Month-Olds from Chocolate Bars, Cookies, Infant Cereals, and Chocolate Drinks in Brazil.

The use of fluoride (F) in the prevention of dental caries is established. However, a high amount of F intake during tooth development can cause dental fluorosis The aim of this study was to analyze variations in F concentrations in chocolate bars (CB), chocolate cookies (CC), infant cereals (IC), and chocolate milk drinks (CD) to determine the daily intake of F from different sources by children at the age of risk for developing dental fluorosis. Distinct brands of CB, CC, IC, and CD were analyzed. Fluoride was separated by hexamethyldisiloxane-facilitated diffusion. Analysis was made in triplicate with an F ion-specific electrode. F ingestion (mg/kg body weight) was evaluated with the suggested consumption (0.05-0.07 mg/kg/day) for children aged 24 months (12 kg). The concentrations for all the analyzed products ranged from 0.025 to 1.827 µg/g F. The mean (range) F concentrations were CB= 0.210 ± 0.205 µg/g (0.073-0.698, n = 8), CC = 0.366 ± 0.416 µg/g (0.320-1.827, n = 9), IC = 0.422 ± 0.395 µg/g (0.073-1.061, n = 5), and CD = 0.169 ± 0.170 µg/mL (0.025-0.443, n = 12). The products that had the highest concentration in the categories CB, CC, IC, and CD, respectively, were Nescau-Ball (0.698 µg/g), Passatempo (1.827 µg/g), Milnutri (1.061 µg/g), and Toddynho (0.443 µg/mL). The consumption of only one unit of Toddynho (CD) is equivalent to more than 11% of the maximum suggested daily intake for a 24-month-old child (0.07 mg/kg body weight). When one product from each category is consumed together only once a day, this consumption is equivalent to approximately 24% of the suggested daily intake of fluoride for a 24-month-old child. The presence of high levels of fluoride in certain products suggests that they play a significant role in overall fluoride intake. It is crucial to closely monitor the fluoride content of food and drinks that are consumed by children who are at risk for dental fluorosis, and for product labels to clearly display the fluoride concentrations.

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.

Acquired pellicle protein-based engineering protects against erosive demineralization.

OBJECTIVES: To evaluate, in vivo: 1) proteomic alterations in the acquired enamel pellicle (AEP) after treatment with sugarcane-derived cystatin (CaneCPI-5), hemoglobin (HB), statherin-derived peptide (StN15) or their combination before the formation of the AEP and subsequent erosive challenge; 2) the protection of these treatments against erosive demnineralization. MATERIALS AND METHODS: In 5 crossover phases, after prophylaxis, 10 volunteers rinsed (10 mL, 1 min) with: deionized water-1, 0.1 mg/mL CaneCPI-5-2, 1.0 mg/mL HB-3, 1.88 × 10-5 M StN15-4 or their combination-5. AEP was formed (2 h) and enamel biopsy (10 µL, 1%citric acid, pH 2.5, 10 s) was performed on one incisor for calcium analysis. The same acid was applied on the vestibular surfaces of the remaining teeth. The acid-resistant proteins within the remaining AEP were collected. Samples were quantitatively analyzed by label-free proteomics. RESULTS: Treatment with the proteins/peptide, isolated or combined, increased several acid-resistant proteins in the AEP, compared with control. The highest increases were seen for PRPs (32-fold, StN15), profilin (15-fold, combination), alpha-amylase (9-fold; StN15), keratins (8-fold, CaneCPI-5 and HB), Histatin-1 (7-fold, StN15), immunoglobulins (6.5-fold, StN15), lactotransferrin (4-fold, CaneCPI-5), cystatins, lysozyme, protein S-100-A9 and actins (3.5-fold, StN15), serum albumin (3.5-fold, CaneCPI-5 and HB) and hemoglobin (3-fold, StN15). Annexin, calmodulin, keratin, tubulin and cystatins were identified exclusively upon treatment with the proteins/peptide, alone or combined. Groups 2, 3 and 4 had significantly lower Ca released from enamel compared to group 1 (Kruskal-Wallis/Dunn's, p < 0.05). CONCLUSIONS: Treatment with CaneCPI-5, HB or StN15 remarkably increases acid-resistant proteins in the AEP, protecting against erosion. CLINICAL SIGNIFICANCE: Our results show, for the first time, that treatment with proteins/peptide remarkably increases acid-resistant proteins in the AEP, protecting against erosive demineralization. These findings open an avenue for a new preventive approach for erosive demineralization, employing acquired pellicle engineering procedures that may in the future be incorporated into dental products.

Saliva as a diagnostic tool for dental caries, periodontal disease and cancer: is there a need for more biomarkers?

Introduction: A biomarker is a biological indicator of normal or pathogenic processes. Identification of biomarkers is useful for the prevention, diagnosis and prognosis of diseases as well as for monitoring the progression of pathological disorders. Several types of molecules present in biological fluids can act as biomarkers such as DNA, coding and non-coding RNA, lipids, metabolites, proteins and even microbes. In this context, saliva emerges as a useful diagnostic tool for the detection of biomarkers involved with oral and systemic diseases, since it reflects the pathophysiological conditions of the organism and allows early, rapid, practical and noninvasive detection of biomarkers.Areas covered: This review discusses the properties of saliva as a diagnostic tool and addresses the main identified biomarkers related to dental caries, periodontal disease, head and neck cancer and other types of cancer of considerable incidence among the world population.Expert commentary: Despite extensive efforts which have been directed toward the identification of one or a combination of biomarkers with good predictive values for the early detection of dental caries, periodontal disease and cancer, these biomarkers still need validation before chairside point-of-care devices can be widely used in the clinic.

The erosive potential of 1% citric acid supplemented by different minerals: an in vitro study.

PURPOSE: The objective of the present in vitro study was to evaluate the effect of different minerals in combination with 1% citric acid on dental erosion. MATERIALS AND METHODS: Ninety enamel samples were randomly allocated to nine groups (G1: pure 1% citric acid solution [control]; G2: with 1 mM Ca; G3: with 0.047 mM F; G4: with 1 mM Fe; G5: with 1 mM P; G6: with 1 mM Ca and 0.047 mM F; G7: with 1 mM Ca and 1 mM P; G8: with 1 mM Fe and 0.047 mM F; G9: with 1 mM Ca, 1 mM P, 0.047 mM F and 1.0 mM Fe). The samples were subjected to six pH cycles, each consisting of immersion in pure or modified 1% citric acid (1 min) followed by storage in artificial saliva (59 min). Enamel wear was assessed using profilometry. RESULTS: Data were analysed using analysis of variance and Tukey test (P < 0.05). Enamel loss (mean + or - SD) amounted to between 0.87 + or - 0.30 and 1.74 + or - 0.74 microm but did not significantly differ among the groups. CONCLUSIONS: The modification of 1% citric acid with different minerals did not have a protective effect on enamel erosion.

Fluoride intake from regular and low fluoride dentifrices by 2-3-year-old children: influence of the dentifrice flavor.

This study evaluated the fluoride intake from dentifrices with different fluoride concentrations ([F]) by children aged 24-36 months, as well as the influence of the dentifrice flavor in the amount of fluoride ingested during toothbrushing. Thirty-three children were randomly divided into 3 groups, according to the [F] in the dentifrices: G-A (523 microgF/g), G-B (1,062 microgF/g) and G-C (1,373 microgF/g). Dentifrices A and B are marketed for children, while dentifrice C is a regular product. The amount of F ingested was indirectly obtained, subtracting the amount expelled and the amount left on the toothbrush from the amount initially loaded onto the brush. The results were analyzed by ANOVA, Tukey's test and linear regression analysis (p < 0.05). Children ingested around 60% of the dentifrice loaded onto the brush, but no significant differences were seen among the groups (p > 0.05). Mean daily fluoride intake from dentifrice for G-A, G-B and G-C was 0.022(a) feminine, 0.032(a) feminine and 0.061(b) mg F/kg body weight, respectively (p < 0.01). There was a strong positive correlation (r = 0.86, p < 0.0001) between the amount of dentifrice used and the amount of fluoride ingested during toothbrushing. The results indicate the need for instructing children's parents and care givers to use a small amount of dentifrice (< 0.3 g) to avoid excessive ingestion of fluoride. The use of low-[F] dentifrices by children younger than 6 years also seems to be a good alternative to minimize fluoride intake. Dentifrice flavor did not influence the percentage of fluoride intake.

Fluoride intake from regular and low fluoride dentifrices by 2-3-year-old children: influence of the dentifrice flavor

This study evaluated the fluoride intake from dentifrices with different fluoride concentrations ([F]) by children aged 24-36 months, as well as the influence of the dentifrice flavor in the amount of fluoride ingested during toothbrushing. Thirty-three children were randomly divided into 3 groups, according to the [F] in the dentifrices: G-A (523 μgF/g), G-B (1,062 μgF/g) and G-C (1,373 μgF/g). Dentifrices A and B are marketed for children, while dentifrice C is a regular product. The amount of F ingested was indirectly obtained, subtracting the amount expelled and the amount left on the toothbrush from the amount initially loaded onto the brush. The results were analyzed by ANOVA, Tukey's test and linear regression analysis (p < 0.05). Children ingested around 60 percent of the dentifrice loaded onto the brush, but no significant differences were seen among the groups (p > 0.05). Mean daily fluoride intake from dentifrice for G-A, G-B and G-C was 0.022ª, 0.032ª and 0.061b mg F/kg body weight, respectively (p < 0.01). There was a strong positive correlation (r = 0.86, p < 0.0001) between the amount of dentifrice used and the amount of fluoride ingested during toothbrushing. The results indicate the need for instructing children's parents and care givers to use a small amount of dentifrice (< 0.3 g) to avoid excessive ingestion of fluoride. The use of low-[F] dentifrices by children younger than 6 years also seems to be a good alternative to minimize fluoride intake. Dentifrice flavor did not influence the percentage of fluoride intake.

Avaliou-se a ingestão de flúor após uso de dentifrícios contendo diferentes concentrações de flúor ([F]) por crianças entre 24-36 meses de idade, além da influência do sabor do dentifrício na quantidade de flúor ingerida durante a escovação. Dividiram-se 33 crianças aleatoriamente em 3 grupos, de acordo com a [F] nos dentifrícios: G-A (523 μgF/g), G-B (1.062 μgF/g) e G-C (1.373 μgF/g). Os dentifrícios A e B são infantis, e o C, convencional. A quantidade de flúor ingerida foi indiretamente obtida subtraindo-se a quantidade de flúor expelida e a quantidade que restou na escova daquela inicialmente carregada na escova. Os resultados foram analisados por ANOVA, teste de Tukey e análise de regressão linear (p < 0,05). Aproximadamente 60 por cento do dentifrício carregado na escova foi ingerido pelas crianças, embora sem diferenças significativas entre os grupos (p > 0,05). A ingestão média diária de flúor para G-A, G-B e G-C foi 0,022ª, 0,032ª e 0,061b mg F/kg de peso corporal, respectivamente (p < 0,01). Houve uma forte correlação positiva (r = 0,86, p < 0,0001) entre a quantidade de dentifrício utilizada e a quantidade de flúor ingerida durante a escovação. Os resultados indicam a necessidade de se instruir pais e cuidadores de crianças a utilizarem uma quantidade pequena de dentifrício (< 0,3 g) para se evitar ingestão excessiva de flúor. O uso de dentifrícios com [F] reduzida por crianças menores de 6 anos também se constitui numa boa alternativa para se minimizar a ingestão de flúor. O sabor do dentifrício não influenciou na porcentagem de ingestão deste íon.