Responses of Dental Pulp Cells to a Less Invasive Bleaching Technique Applied to Adhesively Restored Teeth

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Effect of hydrogen peroxide concentration on enamel color and microhardness

Objectives: The aim of this study was to investigate the effect of hydrogen peroxide gels with different concentrations (20%, 25%, 30%, and 35%) on enamel Knoop microhardness (KNIT) as well as on changes in dental color (C).Methods: Cylindrical specimens of enamel/dentin (3-nun diameter and 2-nun thickness) were obtained from bovine incisors and randomly divided into six groups (n=20), according to the concentration of the whitening gel (20%, 25%, 30%, 35%, control, thickener). After polishing, initial values of KNH0 and color measurement, assessed by spectrophotometry using the CIE L*a*b* system, were taken from the enamel surface. The gels were applied on the enamel surface for 30 minutes, and immediate values of KNHi were taken. After seven days of being stored in artificial saliva, new measures of KNH7 and color (L-7* a(7)* b(7)*, for calculating Delta E, Delta L, and Delta b) were made. Data were submitted to statistical analysis of variance, followed by Tukey test (p<0.05).Results: Differences in gel concentration and time did not influence the microhardness (p=0.54 and p=0.29, respectively). In relation to color changes, Delta E data showed that the 35% gel presented a higher color alteration than the 20% gel did (p=0.006).Conclusion: Bleaching with 35% hydrogen peroxide gel was more effective than with the 20% gel, without promoting significant adverse effects on enamel surface microhardness.

In vitro effect of calcium-containing prescription-strength fluoride toothpastes on bovine enamel erosion under hyposalivation-simulating conditions

Purpose: To evaluate the ability of calcium-containing prescription-strength fluoride (F) toothpastes in preventing enamel erosion under low salivary flow simulating conditions. Methods: Enamel and dentin bovine specimens were assigned to the following groups: A - placebo; B - 1,100 ppm F/NaF (Aquafresh Advanced); C - 5,000 ppm F/NaF (Prevident 5000 Booster); D - 5000 ppm F/NaF+calcium sodium phosphosilicate (Topex Renew); and E - 5,000 ppm F/NaF+tri-calcium phosphate (Clinpro 5000). Specimens were positioned in custom-made devices, creating a sealed chamber on the surface, connected to peristaltic pumps. Citric acid was injected into the chamber for 2 minutes, followed by artificial saliva (0.05 ml/minute), for 60 minutes, 4x/day, for 3 days. Aquafresh was also tested under normal salivary flow (0.5 ml/minute), as reference (Group F). Specimens were exposed to the toothpastes for 2 minutes, 2x/day. After cycling, surface loss (SL) and concentration of loosely- and firmly-bound F were determined. Data were analyzed by ANOVA. Results: Group A (placebo) presented highest surface loss (SL), while Group F had the lowest, for both substrates. For enamel, none of the dentifrices differed from Group B or among each other. For dentin, none of the dentifrices differed from Group B, but Group E showed greater protection than Group C. Group E presented the highest F concentrations for both substrates, only matched by Group D for firmly-bound fluoride on enamel. All fluoridated dentifrices tested reduced SL, with no additional benefit from higher F concentrations. Some formulations, especially Clinpro 5000, increased F availability on the dental substrates, but no further erosion protection was observed.

Evaluation of the bleached human enamel by scanning electron microscopy

Since bleaching has become a popular procedure, the effect of peroxides on dental hard tissues is of great interest in research. Purpose: The aim of this in vitro study was to perform a qualitative analysis of the human enamel after the application of in-office bleaching agents, using Scanning Electron Microscopy (SEM). Materials and Methods: Twenty intact human third molars extracted for orthodontic reasons were randomly divided into four groups (n=5) treated as follows: G1- storage in artificial saliva (control group); G2- four 30-minute applications of 35% carbamide peroxide (total exposure: 2h); G3- four 2-hour exposures to 35% carbamide peroxide (total exposure: 8h); G4- two applications of 35% hydrogen peroxide, which was light-activated with halogen lamp at 700mW/cm2 during 7min and remained in contact with enamel for 20min (total exposure: 40min). All bleaching treatments adopted in this study followed the application protocols advised by manufacturers. Evaluation of groups submitted to 35% carbamide peroxide was carried out after two time intervals (30 minutes and 2 hours per session), following the extreme situations recommended by the manufacturer. Specimens were prepared for SEM analysis performing gold sputter coating under vacuum and were examined using 15kV at 500x and 2000x magnification. Results: Morphological alterations on the enamel surface were similarly detected after bleaching with either 35% carbamide peroxide or 35% hydrogen peroxide. Surface porosities were characteristic of an erosive process that took place on human enamel. Depression areas, including the formation of craters, and exposure of enamel rods could also be detected. Conclusion: Bleaching effects on enamel morphology were randomly distributed throughout enamel surface and various degrees of enamel damage could be noticed. Clinical significance: In-office bleaching materials may adversely affect enamel morphology and therefore should be used with caution.

Dental mineralization and salivary activity are reduced in offspring of spontaneously hypertensive rats (SHR)

Several pathologies have been diagnosed in children of hypertensive mothers; however, some studies that evaluated the alterations in their oral health are not conclusive. This study analyzed the salivary gland activity and dental mineralization of offsprings of spontaneously hypertensive rats (SHR). Thirty-day-old SHR males and Wistar rats were studied. The salivary flow was evaluated by injection of pilocarpine, the protein concentration and salivary amylase activity, by the Lowry method and kinetic method at 405 nm, respectively. Enamel and dentin mineralization of the mandibular incisors was quantified with aid of the microhardness meter. The results were analyzed by the ANOVA or Student's t test (p<0.05). It was noticed that the salivary flow rate (0.026 mL/min/100 g ± 0.002) and salivary protein concentration (2.26 mg/mL ± 0.14) of SHR offspring were reduced compared to Wistar normotensive offspring (0.036 mL/min/100 g ± 0.003 and 2.91 mg/mL ± 0.27, respectively), yet there was no alteration in amylase activity (SHR: 242.4 U/mL ± 36.9; Wistar: 163.8 U/mL ± 14.1). Microhardness was lower both in enamel (255.8 KHN ± 2.6) and dentin (59.9 KHN ± 0.8) for the SHR teeth compared to the Wistar teeth (enamel: 328.7 KHN ± 3.3 and dentin: 67.1 KHN ± 1.0). These results suggest that the SHR offspring are more susceptible to development of pathologies impairing oral health, once they presented lesser flow and salivary protein concentration and lower dental mineralization.

Effect of incorporation of remineralizing agents into bleaching gels on the microhardness of bovine enamel in situ

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

In vitro effect of sodium trimetaphosphate additives to conventional toothpastes on enamel demineralization

The aim of this study was to evaluate the ability of conventional toothpastes (1100 ppm F) supplemented with sodium trimetaphosphate (TMP) in demineralization. Blocks of enamel were selected and then divided into seven experimental groups of 12: toothpaste without F and TMP (placebo), toothpaste with 1100 ppm F (1100), and toothpaste with 1100 ppm F supplemented with TMP-1 % (1100 1 % TMP), 3 % (1100 3 % TMP), 4.5 % (1100 4.5 % TMP), 6 % (1100 6 % TMP), and 9 % (1100 9 % TMP). Blocks were subjected to five pH cycles (demineralizing/remineralizing solutions) at 37 °C and treated with toothpaste slurries twice daily, after which the blocks were maintained for 2 days in fresh remineralizing solution. Following treatments, surface hardness (SHf) and cross-sectional hardness were determined for calculating the integrated loss of subsurface hardness (ΔKHN). The fluoride present in the enamel was also measured. The SHf and ΔKHN measurements showed that supplementation with 3 % TMP was the most effective (p < 0.001) and showed greater concentration of F in the enamel (p < 0.001). Addition of 3 % TMP to a conventional toothpaste (1100 ppm F) showed greater efficacy in reducing enamel demineralization. Fluoride toothpastes containing trimetaphosphate possess good anticaries potential required to reduce the prevalence of dental caries in high-risk patients.

Influence of whitening gel application protocol on dental color change

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Enamel microabrasion: an overview of clinical and scientific considerations

Superficial stains and irregularities of the enamel are generally what prompt patients to seek dental intervention to improve their smile. These stains or defects may be due to hypoplasia, amelogenesis imperfecta, mineralized white spots, or fluorosis, for which enamel microabrasion is primarily indicated. Enamel microabrasion involves the use of acidic and abrasive agents, such as with 37% phosphoric acid and pumice or 6% hydrochloric acid and silica, applied to the altered enamel surface with mechanical pressure from a rubber cup coupled to a rotatory mandrel of a low-rotation micromotor. If necessary, this treatment can be safely combined with bleaching for better esthetic results. Recent studies show that microabrasion is a conservative treatment when the enamel wear is minimal and clinically imperceptible. The most important factor contributing to the success of enamel microabrasion is the depth of the defect, as deeper, opaque stains, such as those resulting from hypoplasia, cannot be resolved with microabrasion, and require a restorative approach. Surface enamel alterations that result from microabrasion, such as roughness and microhardness, are easily restored by saliva. Clinical studies support the efficacy and longevity of this safe and minimally invasive treatment. The present article presents the clinical and scientific aspects concerning the microabrasion technique, and discusses the indications for and effects of the treatment, including recent works describing microscopic and clinical evaluations.

Amorphous calcium phosphate sealants: remineralization of carious lesions in the enamel can be achieved with use of pit-and-fissure sealants containing amorphous calcium phosphate

New pit-and-fissure sealants with the capacity to release calcium and phosphate because of the presence of ACP have been introduced into the dental marketplace. With the continuous introduction of new dental materials, it is important not only to research and confirm their properties, but also to propose modifications or associations that may contribute to their improvement.

Influência da espessura do substrato dental sobre a eficácia clareadora e citotoxicidade de diferentes protocolos de clareamento

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Comparative clinical study of the effectiveness of different dental bleaching methods - two year follow-up

This study evaluated color change, stability, and tooth sensitivity in patients submitted to different bleaching techniques. Material and methods: In this study, 48 patients were divided into five groups. A half-mouth design was conducted to compare two in-office bleaching bleaching techniques (with and without light activation): G1: 35% hydrogen peroxide (HP) (Lase Peroxide - DMC Equipments, Sao Carlos, SP, Brazil) + hybrid light (HL) (LED/Diode Laser, Whitening Lase II DMC Equipments, Sao Carlos, SP, Brazil); G2: 35% HP; G3: 38% HP (X-traBoost - Ultradent, South Jordan UT, USA) + HL; G4: 38% HP; and G5: 15% carbamide peroxide (CP) (Opalescence PF - Ultradent, South Jordan UT, USA). For G1 and G3, HP was applied on the enamel surface for 3 consecutive applications activated by HL. Each application included 3x3' HL activations with 1' between each interval; for G2 and G4, HP was applied 3x15' with 15' between intervals; and for G5, 15% CP was applied for 120'/10 days at home. A spectrophotometer was used to measure color change before the treatment and after 24 h, 1 week, 1, 6, 12, 18 and 24 months. A VAS questionnaire was used to evaluate tooth sensitivity before the treatment, immediately following treatment, 24 h after and finally 1 week after. Results: Statistical analysis did not reveal any significant differences between in-office bleaching with or without HL activation related to effectiveness; nevertheless the time required was less with HL. Statistical differences were observed between the result after 24 h, 1 week and 1, 6, 12, 18 and 24 months (integroup). Immediately, in-office bleaching increased tooth sensitivity. The groups activated with HL required less application time with gel. Conclusion: All techniques and bleaching agents used were effective and demonstrated similar behaviors.

Probing the Enamel Topography After Acid Erosion by Scanning Electrochemical Microscopy

In this work, we present an investigation on the thickness of the eroded enamel layer in tooth samples after exposure to citric and hydrochloric acid by using Scanning Electrochemical Microscopy (SECM). Approaching curves with typical negative feedback behavior were obtained in enamel samples for evaluation of topographic changes. In a control experiment, SECM images showed no significant difference in the current monitored during the scan, implying that enamel demineralization did not occur in mineral water medium. Topographic SECM images obtained after contact with citric and hydrochloric acid for different periods of time showed a significant increase in the current relative to a previously protected surface, indicating the structural loss of enamel. The thickness of the enamel layer eroded after contact with hydrochloric acid was significantly higher when compared to the one obtained with citric acid. Hence, our results showed that the enamel acid erosion is a relatively fast process, which is strongly dependent on parameters such as pH, time, acid strength and acid concentration.

Quantitative Proteomic Analysis of the Effect of Fluoride on the Acquired Enamel Pellicle

The acquired enamel pellicle (AEP) is a thin film formed by the selective adsorption of salivary proteins onto the enamel surface of teeth. The AEP forms a critical interface between the mineral phase of teeth (hydroxyapatite) and the oral microbial biofilm. This biofilm is the key feature responsible for the development of dental caries. Fluoride on enamel surface is well known to reduce caries by reducing the solubility of enamel to acid. Information on the effects of fluoride on AEP formation is limited. This study aimed to investigate the effects of fluoride treatment on hydroxyapatite on the subsequent formation of AEP. In addition, this study pioneered the use of label-free quantitative proteomics to better understand the composition of AEP proteins. Hydroxyapatite discs were randomly divided in 4 groups (n = 10 per group). Each disc was exposed to distilled water (control) or sodium fluoride solution (1, 2 or 5%) for 2 hours. Discs were then washed and immersed in human saliva for an additional 2 hours. AEP from each disc was collected and subjected to liquid chromatography electrospray ionization mass spectrometry for protein identification, characterization and quantification. A total of 45 proteins were present in all four groups, 12 proteins were exclusively present in the control group and another 19 proteins were only present in the discs treated with 5% sodium fluoride. Relative proteomic quantification was carried out for the 45 proteins observed in all four groups. Notably, the concentration of important salivary proteins, such as statherin and histatin 1, decrease with increasing levels of fluoride. It suggests that these proteins are repulsed when hydroxyapatite surface is coated with fluoride. Our data demonstrated that treatment of hydroxyapatite with fluoride (at high concentration) qualitatively and quantitatively modulates AEP formation, effects which in turn will likely impact the formation of oral biofilms.

Xylitol concentrations in artificial saliva after application of different xylitol dental varnishes

Objective: The present study analyzed xylitol concentrations in artificial saliva over time after application of varnishes containing 10% and 20% xylitol. Material and Methods: Fifteen bovine enamel specimens (8x4 mm) were randomly allocated to 3 groups (n=5/group), according to the type of varnish used: 10% xylitol, 20% xylitol and no xylitol (control). After varnish application (4 mg), specimens were immersed in vials containing 500 mu L of artificial saliva. Saliva samples were collected in different times (1, 8, 12, 16, 24, 48 and 72 h) and xylitol concentrations were analyzed. Data were assessed by two-way repeated-measures ANOVA (p<0.05). Results: Colorimetric analysis was not able to detect xylitol in saliva samples of the control group. Salivary xylitol concentrations were significantly higher up to 8 h after application of the 20% xylitol varnish. Thereafter, the 10% xylitol varnish released larger amounts of that polyol in artificial saliva. Conclusions: Despite the results in short-term, sustained xylitol releases could be obtained when the 10% xylitol varnish was used. These varnishes seem to be viable alternatives to increase salivary xylitol levels, and therefore, should be clinically tested to confirm their effectiveness.