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Mercury in solution following exposure of various amalgams to carbamide peroxides.

Carbamide peroxide (CP) is an easily administered material for whitening teeth. Although toxicological research on CP alone has revealed no adverse health effects, possible oxidation and release of mercury from amalgams have not previously been investigated. This research evaluated the quantitative release of mercury from amalgams into solution by CP. CP preparations can generally be divided into two classes based on the presence or absence of carbopol, an oxygen-releasing inhibitor. Rembrandt (R), a 10% CP with carbopol and White and Brite (WB), a 10% CP without carbopol were used in this study. Four different types of amalgams [Dispersalloy (D), Sybraloy (S), Tytin (T) and Valiant Ph.D. (V)] were selected. Uniform samples of the four amalgams were prepared and stored at 37 degrees C for 1 week. Vials of saline (10 ml), R and WB were prepared. R and WB were mixed with saline to a 50:50 solution to reduce viscosity and facilitate stirring. Magnetic teflon coated stir bars were placed in all vials, and one amalgam specimen was placed in each non-control vial. After being stirred for 8 hours, solutions were analyzed for elemental mercury content using a Jerome Gold Film Mercury Analyzer. All background mercury levels were zero, but following the experiment there were significantly higher amounts of mercury in the CP solutions as compared to the 100% saline solutions. These results suggest there is an active oxidation of the amalgam releasing mercury ions into solution.

Acrylic Resins↗

At-home vital bleaching: a comparison of hydrogen peroxide and carbamide peroxide treatments.

Tray bleaching of vital teeth performed at home by the patient under the dentist s supervision, whether alone or in combination with any of the in-office techniques, provides an interesting alternative to other methods employed in this type of dental treatment. This bleaching procedure applies low-concentration peroxides to the enamel by means of a custom-made mouth tray specifically designed for this purpose. The aim of this study is to examine and compare two commercially-available bleaching products, at equivalent concentrations, for use in this technique: VivaStyle (Vivadent) and FKD (Kin); the former is a 10% carbamide peroxide and the latter a 3.5% hydrogen peroxide formulation. It examines the parameters that must be monitored during the application of this type of procedure and presents 6 cases (3 treated with one of the above-mentioned products and the other 3 with the other), establishing the bleaching power of the products and the appearance and intensity of post-operatory hypersensitivity. The results obtained show that both products are effective for the purpose for which they were designed. In general, dental hypersensitivity was minimal.

Carbamide Peroxide↗

Effects of two 10% peroxide carbamide bleaching agents on dentin microhardness at different time intervals.

OBJECTIVE: The purpose of this in vitro study was to evaluate the microhardness of human dentin exposed to two 10% carbamide peroxide agents at different bleaching times. METHOD AND MATERIALS: Opalescence 10% and Rembrandt 10% were tested. A placebo agent was used as a control group. The bleaching and placebo agents were applied to the surface of human dentin fragments for 8 hours and then stored in individual receptacles with artificial saliva for the remaining 16 hours each day. Microhardness testing was performed at baseline, after 8 hours, 7, 14, 21, 28, 35, and 42 days of treatment, and 7 and 14 days posttreatment. RESULTS: Analysis of Variance and the Tukey test revealed significant differences in microhardness values for dentin treated with the agents within each time interval. There was a decrease in the microhardness values of dentin for both bleaching agents after 8 hours of treatment. Fourteen days after the completion of treatment, the microhardness values for dentinal surfaces treated with either Opalescence or Rembrandt reached the baseline values; the dentinal surfaces treated with a placebo exhibited an increase in microhardness values posttreatment. CONCLUSION: Ten percent carbamide peroxide bleaching agents decreased dentinal microhardness over time, but after 14 days in artificial saliva storage at the completion of treatment, the baseline microhardness values were recovered.

Analysis of Variance↗

Scanning electron microscopy study of dental enamel surface exposed to 35% hydrogen peroxide: alone, with saliva, and with 10% carbamide peroxide.

Several vital bleaching systems have been introduced in response to the demand in esthetic dentistry. The active agents are commonly hydrogen peroxide or carbamide peroxide used in at-home or in-office techniques. Although generally positive results have been reported concerning the whitening ability of these agents, concerns still remain as to their effects on dental tissues. The purpose of this investigation was to evaluate the effect of these bleaching agents on the enamel surface morphology. Twelve extracted teeth were used according to three experimental protocols. In experimental protocol 1, specimens were treated with 35% hydrogen peroxide. In experimental protocol 2, after treatment with 35% hydrogen peroxide the specimens were immersed in natural saliva for 1 week. In experimental protocol 3, 35% hydrogen peroxide was applied once and 10% carbamide peroxide was applied for 1 week (12 h of 10% carbamide peroxide alternating with 12 h saliva). Scanning electron microscopy evaluation revealed that regional variation in tooth morphology surface sometimes exceeded the effects of the peroxide used according to experimental protocols. Thirty-five percent hydrogen peroxide had a tendency to promote an increase in density of pits. Precipitates were observed on specimen surfaces immersed in natural saliva according to protocol 2. A smooth and shiny surface was observed in specimens treated according to protocol 3. The potential relationship between surface alterations and differences in enamel permeability is currently under investigation. CLINICAL SIGNIFICANCE The differences in various articles written on the subject cannot be reconciled because of the lac of standardization of baseline data regarding factors such as location on the tooth, type of tooth eruption or noneruption, and age in the oral cavity. This article demonstrates that, despite changes observed in the enamel surface after bleaching, normal variation in tooth morphology may exceed the effects of 35% hydrogen peroxide and 10% carbamide peroxide on the teeth. Hence, considering the morphologic features of the tooth surface, bleaching, as described in thi study, can be considered safe for enamel.

Adult↗

Oxidative stress aspects of the cytotoxicity of carbamide peroxide: in vitro studies.

Carbamide peroxide is the active ingredient in many at-home patient-applied tooth whiteners. The cytotoxicity of carbamide peroxide, as related to oxidative stress, was evaluated in vitro with several human cell lines, including Smulow-Glickman (S-G) gingival epithelial cells. The potency of carbamide peroxide was related to its hydrogen peroxide component rather than to carbamide, was eliminated in the presence of exogenous catalase, and was enhanced in the presence of aminotriazole, an inhibitor of cellular catalase. The intracellular level of glutathione, a scavanger of toxic oxygen metabolites, was decreased in cells exposed to carbamide peroxide; at higher concentrations of carbamide peroxide, leakage of lactic acid dehydrogenase was also evident. Cells pretreated with the glutathione-depleting agents, buthionine sulfoximine, chlorodinitrobenzene, and bis(chloroethyl) nitrosourea, were hypersensitive to subsequent challenge with carbamide peroxide. Conversely, pretreatment with the iron chelator, deferoxamine, protected the cells against subsequent exposure to carbamide peroxide.

Carbamide Peroxide↗

Safety issues when using carbamide peroxide to bleach vital teeth--a review of the literature.

Hydrogen Peroxide is used to bleach discoloured teeth but since its introduction in the late nineteenth century there have been concerns about its safety and efficacy. This paper reviews the literature on hydrogen peroxide and carbamide peroxide and assesses if these products can be recommended for clinical use. The authors used a Medline search to find the literature for review and from these the findings were divided into laboratory, animal and human studies. In conclusion no dental treatment is without risk but from the evidence it seems that bleaching teeth is comparatively safe.

Animals↗

Effects of three concentrations of carbamide peroxide on the structure of enamel.

Carbamide peroxide bleaching agents have raised important questions on their potential adverse effects on the structure of enamel. The purpose of this study was to examine the effects of three carbamide peroxide bleaching agents in different concentrations (10, 16 and 35%) on the structure of enamel. Forty enamel slabs prepared from human third molars were divided equally among four groups. The specimens in the first and second group were subjected to 10% or 16% carbamide peroxide for 8 h per day for 6 weeks. Thirty-five percent carbamide peroxide was applied to the third group for 30 min a day for a 4 day period according to the manufacturers recommendation. The fourth group served as the control and was kept in artificial saliva during the test period. At the completion of the treatment, all the specimens were examined by infrared absorbtion spectroscopy and X-ray diffraction analysis. The results revealed that 10% or 16% carbamide peroxide did not seem to effect the structure of enamel, whereas 35% carbamide did affect the structure. The use of lower concentrations of carbamide peroxide (10-16%) is recommended over higher concentrations (35%) to avoid changes to the enamel.

Carbamide Peroxide↗

Higher-concentration carbamide peroxide effects on surface roughness of composites.

BACKGROUND: Ten percent carbamide peroxide has been used extensively within the dental profession for the purpose of bleaching teeth. Although this method has been successful, the use of higher-concentration carbamide peroxides (20-35%) for home bleaching has increased substantially in the past few years. The purpose of this study was to evaluate changes in surface roughness of a hybrid and a microfilled composite after exposure to bleaching agents containing higher concentrations of carbamide peroxide. MATERIALS AND METHODS: Sixty-eight circular, resin-based composite specimens (8 mm x 2 mm) were prepared against a Mylar surface. Half of the specimens were fabricated from a hybrid composite material and the other half with a microfilled composite. Specimens were polished with aluminum oxide finishing disks, divided into four groups, and then exposed either to 20% carbamide peroxide for 3 hours per day for 14 days or to 35% carbamide peroxide for 1 hour per day for 14 days. The mean surface roughness (Ra) was determined for each specimen before and after exposure to bleaching agents, using mechanical profilometry. Data were analyzed by two-way analysis of variance (ANOVA) for differences between composite type, carbamide peroxide concentrations, and interaction of the two factors with respect to average surface roughness. All data analysis was performed at alpha = 0.05. RESULTS: Mean change from baseline of surface roughness (Ra) of microfilled composite and 20% carbamide peroxide was -0.03 microm +/- 0.10 microm; microfilled composite and 35% carbamide peroxide: 0.01 microm x 0.02 microm; hybrid composite and 20% carbamide peroxide: -0.43 microm x 0.91 microm; hybrid composite and 35% carbamide peroxide: -0.19 microm x 0.43 microm. Surface roughness of hybrid or microfilled composite did not change significantly from baseline with either concentration of carbamide peroxide (p = .300). CLINICAL SIGNIFICANCE: Higher-concentration carbamide peroxide bleaching agents, used as intended by the manufacturer, pose no significant risk to resin composite restoration surfaces.

Aluminum Oxide↗

A randomized clinical trial comparing a novel 5.3% hydrogen peroxide whitening strip to 10%, 15%, and 20% carbamide peroxide tray-based bleaching systems.

A randomized and controlled, parallel-group clinical trial compared the whitening benefits of a novel 5.3% hydrogen peroxide bleaching strip to 10%, 15%, and 20% carbamide peroxide tray-based bleaching systems. A total of 36 healthy adults were randomized to a 14-day regimen in which both arches were whitened for 1 hour per day in the whitening-strip group or 2 hours per day in the tray groups. Efficacy was measured objectively using digital images of the anterior teeth at baseline and after 14 days of treatment. Overall tooth color (L*a*b*) was derived from individual pixel values, and then mean levels of delta b*, delta L*, and composite color (delta E*) were compared using analysis of covariance. After 14 days of treatment, all groups experienced a greater than 1-unit mean improvement in delta b*, delta L*, and delta E* relative to baseline. For the primary study variable, reduction of yellow (delta b*) outcomes after 14 hours of using the experimental strip were comparable to those observed with the 10% tray group after 28 hours of use. These two treatment groups did not differ statistically with respect to any of the color measurements used in this study. For the tray groups, there was a reasonable dose relationship for the primary end point, delta b*, with the 15% and 20% tray groups averaging 17% and 68% improvements in yellow, respectively, over the 10% group. Except for the 20% carbamide peroxide system, where sensitivity was relatively common, all test products were well tolerated. In this first comparative evaluation vs marketed controls, use of the whitening strips twice daily for 14 days yielded a highly significant improvement in tooth color vs baseline.

Adult↗

Changes in surface levels of mercury, silver, tin, and copper of dental amalgam treated with carbamide peroxide and hydrogen peroxide in vitro.

OBJECTIVES: The effect of 10% carbamide peroxide or 10% hydrogen peroxide on the surface levels of mercury, silver, tin, and copper of amalgam fillings was tested in vitro with scanning electron microscopy and energy dispersive spectrometric microanalysis. STUDY DESIGN: Samples of amalgam were treated for 14 and 28 days with either 10% carbamide peroxide or 10% hydrogen peroxide solutions and compared with phosphate buffer controls. RESULTS: A significant increase in mercury levels occurred after treatment with carbamide peroxide for 14 days (p < 0.01) and 28 days (p < 0.001) and after treatment with hydrogen peroxide for 28 days (p < 0.001). A significant increase in silver levels occurred after treatment with carbamide peroxide for 14 days (p < 0.05) and 28 days (p < 0.01) and subsequent to treatment with hydrogen peroxide for 14 days (p < 0.05) and 28 days (p < 0.001). A significant reduction in tin levels occurred after treatment with hydrogen peroxide for 14 days (p < 0.01) and 28 days (p < 0.001), and after treatment with carbamide peroxide for 28 days (p < 0.01). A significant reduction in copper levels was found after treatment with carbamide peroxide for 14 days (p < 0.05). CONCLUSIONS: It appears that prolonged treatment with bleaching agents may cause microstructural changes in amalgam surfaces, possibly increasing exposure of patients to toxic byproducts.

Carbamide Peroxide↗

Mercury release from dental amalgam after treatment with 10% carbamide peroxide in vitro.

OBJECTIVES: The effect of 10% carbamide peroxide on mercury release from dental amalgams was assessed in vitro by using a cold-vapor atomic absorption Mercury Analyzer System. STUDY DESIGN: Samples of 4 commercial brands of dental amalgam, Megaloy (Dentsply/Caulk, Milford, Del), Mega+ (CFPM, Aulnaye, France), Nongama 2 (Silmet, Or Yehuda, Israel), and Valiant Ph.D. (Dentsply/Caulk, Milford, Del), were treated for 48 hours with 10% carbamide peroxide and compared with samples treated with phosphate buffer. RESULTS: Amalgam specimens exposed for 48 hours to 10% carbamide peroxide showed significantly higher concentrations of mercury in solution as compared with specimens treated with phosphate buffer (P <.001). Megaloy and Valiant Ph.D. yielded significantly higher mercury concentrations in solution than Mega+ and Nongama 2 (P <.001). Mega+ yielded significantly higher mercury concentrations in solution than Nongama 2 (P <.05). No significant differences were found in mercury concentrations in solution between Megaloy and Valiant Ph.D. CONCLUSIONS: Treatment with 10% carbamide peroxide bleaching agents caused an increase in mercury release from amalgam restorations, possibly increasing exposure of patients to its adverse effects. Amalgam brands differed in the amounts of mercury release after bleaching with carbamide peroxide.

Carbamide Peroxide↗

Safety issues when using a 16% carbamide peroxide whitening solution.

BACKGROUND: The scientific literature is lacking on the occurrence of side effects and other safety issues when using carbamide peroxide whitening solutions of concentrations greater than 10%. This double-blind nightguard vital bleaching study compares safety issues when using 16% carbamide peroxide against a placebo or 10% carbamide peroxide (Nite White Classic by Discus Dental Inc.). Evaluated were changes in gingival index, plaque index, nonmarginal gingival index, nongingival oral mucosal index, tooth vitality, and the patients' perceptions of tooth sensitivity and gingival irritation. MATERIALS AND METHODS: Twenty female dental hygiene students participated in the study. Each participant wore a maxillary treatment tray for 1 week without any solution and then for 8 to 10 hours per night for 14 nights, filling each quadrant with placebo, 10% carbamide peroxide, or 16% carbamide peroxide, using a split tray design. RESULTS: With respect to gingival index, plaque index, nonmarginal gingival index, nongingival oral mucosa index, tooth vitality, and tooth sensitivity, there were no statistically significant differences between the 16% carbamide peroxide solution and the other two solutions (p > .05). Quadrants receiving the 16% carbamide peroxide solution experienced more gingival irritation than quadrants receiving placebo or 10% carbamide peroxide solution (p > .05). CONCLUSIONS: When evaluating the above-mentioned safety issues, except for gingival irritation, there were no statistically significant differences between a 16% carbamide peroxide solution and 10% carbamide peroxide solution or a placebo when used as described here. CLINICAL SIGNIFICANCE: Among the 20 participants whose data were analyzed, it was found that a 16% carbamide peroxide whitening solution (Nite White Classic), when used as described in this study, can be effective in nightguard vital bleaching with no statistical differences in gingival index, plaque index, nonmarginal gingival index, nongingival oral mucosa changes, tooth vitality, or tooth sensitivity, compared with a 10% whitening solution (Nite White Classic). More gingival irritation was experienced with 16% carbamide peroxide. Additionally, 20% of the participants in this study self-reported sensitivity when wearing their treatment tray without any solution, and 36% of the participants reported sensitivity to the placebo solution.

Adult↗

The effect of 10% carbamide peroxide, carbopol and/or glycerin on enamel and dentin microhardness.

This study evaluated the effects of 10% carbamide peroxide, carbopol and glycerin and their associations on microhardness over time on enamel and dentin. Eight treatment agents were evaluated: a commercial bleaching agent containing 10% carbamide peroxide (Opalescence 10% Ultradent), 10% carbamide peroxide, carbopol, glycerin, 10% carbamide peroxide + carbopol, 10% carbamide peroxide + glycerin, carbopol + glycerin and 10% carbamide peroxide + carbopol + glycerin. Three hundred and twenty human dental fragments, 80 sound enamel fragments (SE), 80 demineralized enamel fragments (DE), 80 sound dentin fragments (SD) and 80 demineralized dentin (DD) fragments, were exposed to the treatment agents (n=10). These agents were applied onto the surface of the fragments eight hours a day for 42 days. After eight hours, they were washed from the dental fragment surfaces after five back-and-forth movements with a soft bristle toothbrush under distilled and deionized running water. During the remaining time (16 hours per day), the fragments were kept in individual vials in artificial saliva. After the 42-day treatment period, the specimens were kept individually in artificial saliva for 14 days. Knoop microhardness measurements were performed at baseline, after eight hours, and 7, 14, 21, 28, 35 and 42 days, and 7 and 14 days post-treatment (corresponding to 49 and 56 days after the initial treatment agent applications). The non-parametric Kruskal-Wallis analysis showed significant differences among the agents at each time interval, except at baseline for sound and demineralized enamel and dentin. For SE, SD and DD, there was a decrease in microhardness values during treatment with all agents. There was a tendency towards lower microhardness values after treatment with carbopol and its associations for sound tissues. DD showed low microhardness values during and after treatment with CP and its associations. For DE, there was an increase in microhardness values during treatment with all agents and in the post-treatment phase. The baseline microhardness values were not recovered during the 14-day post-treatment phase. Opalescence 10%, carbamide peroxide, carbopol, glycerin and their associations may change the microhardness of sound and demineralized dental tissues, even in the presence of artificial saliva.

Acrylic Resins↗

Susceptibility of enamel surfaces to demineralization after application of fluoridated carbamide peroxide gels.

The aim of the present study was to evaluate the effect of experimental, fluoridated carbamide peroxide gels on formation of erosively induced demineralization of enamel. Seventy-five bovine enamel specimens were polished for microhardness determination and evenly distributed among 5 groups (A-E). The specimens were treated with 10% carbamide peroxide gel (8 h) and subjected twice to remineralization for 2 h in artificial saliva and demineralisation for 90 s in 1% citric acid, pH 2.2. The cycles of treatment with carbamide peroxide and twofold re- and demineralization were repeated three times. The carbamide peroxide gels were different in pH and fluoride content. Gel A (pH 7.0) and gel B (pH 5.7) were fluoridated (0.5% F), gel C (pH 7.0) and gel D (pH 5.7) were not fluoridated. In control group E the samples were not treated with a gel, but stored in 100% humidity for 8 h instead. Knoop microhardness of the specimens was assessed directly after polishing, and after each carbamide peroxide treatment and demineralization. All specimens showed a loss of microhardness at the end of the experiment. After 3 days, the controls revealed a significantly lower hardness loss compared to the specimens treated with the carbamide peroxide gels. Surface softening was significantly lower for the specimens of group A compared to the remaining groups (B-D), which were not significantly different among each other. It is concluded that treatment with either fluoridated or unfluoridated carbamide peroxide gels, at either neutral and acidic pH, renders enamel more susceptible to demineralization. Use of a fluoridated neutral gel decreases the degree of surface softening compared to the other gels investigated.

Analysis of Variance↗

Effect of carbamide peroxide and hydrogen peroxide on the surface morphology and zinc oxide levels of IRM fillings.

The effect of 10% carbamide peroxide or 10% hydrogen peroxide on the surface morphology and zinc oxide levels of IRM fillings was tested. Ninety IRM samples were treated with either 10% carbamide peroxide, 10% hydrogen peroxide or phosphate buffer which served as control. Treatment consisted of placing the samples in a dry incubator at 37 degrees C for 1, 3 or 7 days. At each time point, the samples were removed from the test solutions, dried and prepared for surface scanning electron microscopy and energy dispersive spectrometric analysis. After 3 days, 10% carbamide peroxide significantly reduced the zinc oxide levels as compared to the 10% hydrogen peroxide group (<0.01) and the controls (p<0.01). 10% hydrogen peroxide reduced the zinc oxide levels similarly to the control. No significant changes in the zinc oxide levels were found between 3 and 7 days in any of the groups tested. Microscopy examination of the carbamide peroxide group revealed granular surface with well defined crystalline areas. In the hydrogen peroxide group, numerous cracks with multiple sun burst-like areas were found. At the macroscopic level, the samples of this group appeared cracked and more swollen, as compared to controls and samples treated with carbamide peroxide. In conclusion, both 10% carbamide peroxide and 10% hydrogen peroxide altered the surface morphology and the zinc oxide levels of IRM fillings, but their modes of action differed.

Carbamide Peroxide↗

Shear bond strength of enamel treated with seven carbamide peroxide bleaching agents.

PURPOSE: Lower average values of bond strength of adhesive systems to enamel bleached with 10% carbamide peroxide agents have been reported, but the effects of higher concentrations of carbamide peroxide bleaching agents are still unknown. The aim of this in vitro study was to evaluate the shear bond strength of enamel treated with different concentrations of carbamide peroxide to an adhesive system after a postbleaching period of storage in artificial saliva for 15 days. MATERIALS AND METHODS: Seven carbamide peroxide bleaching agents with concentrations varying from 10 to 22% were analyzed. A placebo agent was used as a control group. The agents were applied on the enamel fragments for 8 h/d for 42 days. During the remaining time, the specimens were stored in artificial saliva. After that time, the fragments were stored individually in artificial saliva for 15 days. An adhesive system was used to bond resin-based composite cylinders on the enamel surface. Shear bond strength tests were performed and the fractured surfaces of the specimens were visually examined with a stereomicroscope at x 30. RESULTS: The analysis of variance did not show differences in shear bond strength among the treatment agents. The fractures for all treatment agents were predominantly adhesive. CONCLUSION: After 15 days storage in artificial saliva, different concentrations of carbamide peroxide bleaching agents and a placebo agent had the same enamel shear bond strength values. CLINICAL SIGNIFICANCE: Concentrations varying from 10 to 22% carbamide peroxide agents cause no differences in shear bond strength of enamel to an adhesive system after 15 days of storage in artificial saliva.

Analysis of Variance↗

A clinical study of the effect of pellicle on the degradation of 10% carbamide peroxide within the first hour.

OBJECTIVE: The purpose of this study was to determine the degradation of 10% carbamide peroxide within the first hour of use and the effect of pellicle on the degradation of carbamide peroxide in vivo. METHOD AND MATERIALS: Fifteen subjects were assigned to wear nightguard trays filled with bleaching gel for 0.5, 2.5, 5.0, 10.0, 20.0, 40.0, and 60.0 minutes under 2 conditions: (1) subjects did not receive a prophylaxis before each bleaching treatment (pellicle was assumed to be present on the tooth surface); and (2) subjects received prophylaxis to remove pellicle from the tooth surface. At each evaluation time, 3 types of sample were collected: (1) a "grab sample," a sample of gel taken with a spatula from the reservoir of the tray on the maxillary right central incisor; (2) the remaining gel from the tray; and (3) the remaining gel scraped from the teeth. The collected gel samples were analyzed for the amount of peroxide using the United States Pharmacopeia-accepted method. Subjects were asked not to swallow the saliva during treatment but to expectorate into a beaker. The collected saliva was analyzed to determine the amount of peroxide. This sample represented the approximate amount of peroxide that subject would ingest during treatment. RESULTS: Pellicle did not affect the degradation of carbamide peroxide for the teeth or grab samples. The degradation of carbamide peroxide was exponential, except during the first 5 minutes, when the degradation rate was much higher. The saliva collected from subjects during use of the bleaching tray revealed an average of 2.1 mg of carbamide peroxide for 1 hour of bleaching treatment. CONCLUSION: The degradation rate is not affected by the removal of the pellicle.

Analysis of Variance↗