The use of mouthwash containing trimetaphosphate as an adjunct therapy to fluoridated toothpaste reduces enamel demineralization.

INTRODUCTION: The decline in dental caries has been attributed to the widespread use of fluoride (F). Two forms of presentation are fluoridated toothpaste (FT) and mouthwash (MW), widely used by the population. MATERIALS AND METHODS: This study aimed to evaluate in vitro the effects of combining FT and MW, whether supplemented with sodium trimetaphosphate (TMP) or not, on dental enamel demineralization. Bovine enamel blocks (n = 60) were selected based on initial surface hardness (SHi) and divided into 5 experimental groups (n = 12 each): I) Placebo Toothpaste (without F/TMP); II) 1100 ppm F Toothpaste (FT); III) 1100F associated with a MW at 100 ppm F (FT + MW 100F); IV) 1100F associated with a MW at 225 ppm F (FT + MW 250F); and V) 1100F associated with a MW at 100 ppm F supplemented with 0.4 % TMP (FT + MW 100F-TMP). The blocks were treated twice a day, undergoing 5 pH cycles over 7 days. Thus, the percentage change in surface hardness (%SH), integrated subsurface hardness loss (ΔKHN), and the concentration of F, phosphorus (P), and calcium (Ca) in the enamel were determined. The data were submitted to ANOVA and Student-Newman-Keuls test (p < 0.001). RESULTS: The 1100F group was statistically inferior to the groups associated with MW for %SH, ΔKHN, and the concentration of P and Ca in the enamel (p < 0.001). Blocks treated with FT + MW 225F and FT + MW 100F-TMP showed significantly lower %SH compared to the other groups (p < 0.001). The FT + MW 100F - TMP group exhibited the lowest depth mineral loss (ΔKHN), and higher concentration de P in enamel (p < 0.001). CONCLUSION: The adjunct use of MW with FT produces a greater protective effect in inhibiting enamel demineralization, and the supplementation of TMP to the MW with 100F provides a superior effect compared to MW with 225F. CLINICAL SIGNIFICANCE: This combination of treatments could be regarded as one of several alternative fluoride supplements for subjects at elevated risk of caries.

Pomegranate extract in polyphosphate-fluoride mouthwash reduces enamel demineralization.

OBJECTIVES: To evaluate the anti-demineralizing effect of a mouthwash comprising pomegranate peel extract (PPE 3%), sodium trimetaphosphate (TMP 0.3%), and fluoride (F 225 ppm) in an in situ study, and to assess its irritation potential in an ex vivo study. METHODS: This double-blind crossover study was conducted in four phases with 7 days each. Twelve volunteers used palatal appliances containing enamel blocks, which were subjected to cariogenic challenges. The ETF formulation (PPE + TMP + F, pH 7.0), TF formulation (TMP + F, pH 7.0), deionized water (W, pH 7.0), and essential oil commercial mouthwash (CM, 220 ppm F, pH 4.3) were dropped onto the enamel twice daily. The percentage of surface hardness loss, integrated loss of subsurface hardness, calcium, phosphorus, and fluoride in enamel and biofilms were determined. In addition, alkali-soluble extracellular polysaccharide concentrations were analyzed in the biofilms. The irritation potential was evaluated using the hen's egg chorioallantoic membrane test through the vascular effect produced during 300-s of exposure. RESULTS: ETF was the most efficacious in preventing demineralization. It also showed the highest concentrations of calcium and phosphorus in the enamel and in the biofilm, as well as the lowest amount of extracellular polysaccharides in the biofilm. In the eggs, ETF produced light reddening, whereas CM led to hyperemia and hemorrhage. CONCLUSIONS: The addition of PPE to formulations containing TMP and F increased its anti-demineralizing property, and this formulation presented a lower irritation potential than the CM. CLINICAL RELEVANCE: ETF can be a promising alternative alcohol-free mouthwash in patients at high risk of caries.

Effect of nanometric ß-calcium glycerophosphate supplementation in conventional toothpaste on enamel demineralization: An in vitro study.

The aim of this study was to evaluate the effects of supplementing toothpastes containing 1100 ppm F with micrometric or nanometric [beta]-calcium glycerophosphate (ß-CaGPm/ß-CaGPn) on artificial enamel demineralization, using a pH cycling model. Bovine enamel blocks (4 mm × 4 mm, n = 120) selected using initial surface hardness were randomly allocated to ten toothpaste groups (n = 12): without fluoride or ß-CaGPm or ß-CaGPn (Negative control), 1100 ppm F (1100 F), and 1100 ppm F plus 0.125%, 0.25%, 0.5%, and 1.0% of ß-CaGPm or ß-CaGPn. Blocks were treated two times per day with toothpaste slurry and subjected to five pH cycles (demineralizing and remineralizing solutions) at 37 °C. The final surface hardness, percentage of surface hardness loss (%SH), cross-sectional hardness (ΔKHN), and profile analysis and lesion depth subsurface were analysed using polarized light microscopy (PLM). Fluoride (F), calcium (Ca), and phosphorus (P) concentrations were also measured. Data were analysed using ANOVA and Student-Newman-Keuls tests ([alpha] = 0.001). Blocks treated with 1100 F toothpaste containing 0.5%ß-CaGPm or 0.25%ß-CaGPn showed with reduced %SH values when compared with those treated with 1100 F alone (p < 0.001). Reduced lesion depths (ΔKHN and PLM) were observed for the slurry made up of 1100 F and 0.25%ß-CaGPn (p < 0.001). The addition of ß-CaGPm and ß-CaGPn did not influence the enamel F concentration, with the 1100 F/0.25%ß-CaGPn group exhibiting the highest Ca and P enamel concentrations (p < 0.001). Based on the findings of this in vitro study, we can conclude that the fluoride toothpaste produced a superior effect when combined at an appropriate ß-CaGP molar ratio. This effect was achieved with a lower proportion of ß-CaGP in the form of nanometric particles.

Ability of a Blue Hemoglobin-Based Liquid as a Novel Technology to Stain Initial Enamel Demineralization: A Proof-of-Concept in vitro Study.

During orthodontic treatment, enamel demineralization can occur. Its early detection is the basis for efficient preventive measures to arrest or remineralize lesions. In the present study, the application of a novel blue hemoglobin-based liquid (BlueCheck) was evaluated as proof of concept for detection of artificially demineralized smooth surfaces. 60 samples from extracted human posterior teeth were randomly assigned to four groups (15 per group). In 30 of these samples (groups A and B), superficial enamel was removed to create a ground surface. On the surface of other 30 samples (group C and D), orthodontic metal brackets were bonded. On each surface, BC liquid was applied and rinsed with water after 3 min (baseline). All surfaces were checked by two independent observers for presence of blue areas. On each sample, one side was covered by nail varnish to protect this enamel part from demineralization. The samples were demineralized with lactic acid (pH 4.6) for 7 days (group A and C) and 14 days (group B and D), respectively. Mineral loss was determined using quantitative light-induced fluorescence after demineralization. BlueCheck dye was again applied on the samples and evaluated for presence of stained areas. Histological sections were prepared from randomly selected samples and lesion depth was measured. Kruskal-Wallis test was used for group comparison (α = 0.05). After demineralization, median ΔF value for all samples was -8.25% indicating the presence of an initial demineralization. The difference of ΔF values was not statistically significant between samples at 7 or 14 days of demineralization, nor for samples with and without orthodontic brackets (p = 0.13). At baseline, none of the sample surfaces showed discoloration, whereas a distinctive blue color was visible after demineralization in all samples exposed to acid-exposed areas, corresponding to 100% sensitivity. The internal control surfaces (without demineralization) did not show any staining, corresponding to 100% specificity. Histologically measured lesion depths ranged between 200 and 254 µm. In this in vitro study, staining of demineralized enamel surface areas were shown to be reliable. Based on our results, this easily applicable product seems useful to be an adjuvant method to clinical examination to monitor oral health during an orthodontic treatment on tooth surfaces after removal of dental biofilm.

Antimicrobial and cytotoxic capacity of pyroligneous extracts films of Eucalyptus grandis and chitosan for oral applications.

Chitosan films containing distilled pyroligneous extracts of Eucalyptus grandis (DPEC), characterized and developed by Brazilian Agricultural Research Corporation-Embrapa Temperate Agriculture (EMBRAPA-CPACT), were evaluated for antimicrobial activity against Candida albicans, Streptococcus mutans, and Lactobacillus acidophilus by direct contact test. Further, their capacity for the prevention of teeth enamel demineralization and cytotoxicity in vitro were also determined. The natural polymers were tested at different concentrations (1500-7500 µg mL-1) and the formulation of an experimental fluoride varnish with antimicrobial activity was evaluated by direct contact test, whereas cytotoxicity was analyzed through the colorimetric MTT assay. Preliminary data showed no statistically significant differences in cytotoxicity to NIH/3T3 cell line when DPEC is compared to the control group. On the other hand, the antimicrobial capacity and demineralization effects were found between the test groups at the different concentrations tested. Chitosan films containing distilled pyroligneous extracts of E. grandis may be an effective control strategy to prevent biofilm formation related to dental caries when applied as a protective varnish. They may inhibit the colonization of oral microorganisms and possibly control dental caries through a decrease in pH and impairment of enamel demineralization.

Remineralization efficacy of nonfluoride versus herbal-based pediatric dentifrice in demineralized primary teeth.

BACKGROUND: Dental caries is one of the most prevalent posteruptive bacterial infections worldwide, characterized by a progressive demineralization process that affects the mineralized dental tissues. Although the decline of dental caries prevalence can be attributed to the widespread use of dentifrices that contain fluoride, yet there is a need for an advanced alternative nonfluoride remineralizing dentifrice. Yet, there is a need for an advanced alternative nonfluoride remineralizing dentifrice. AIM: The aim of this study was to evaluate and compare the remineralizing effect of nonfluoride-based and herbal-based pediatric dentifrice in demineralized primary teeth with an ideal in vitro method of pH cycling and evaluating the values under Polarized Light Microscope (Olympus BX43) using image analysis software (ProgRes, Speed XT core3). MATERIALS AND METHODS: A total of 30 tooth samples were collected and placed in the demineralizing solution for 96 h to produce a demineralized lesion of approximately 100 µm, and then cut longitudinally into 60 sections that were randomly assigned to two groups with 27 samples each, Group A - nonfluoride-based dentifrice (Mee Mee®), Group B - herbal-based dentifrice (Mamaearth™), after which they were subjected to pH cycling for 7 days along with dentifrice slurry preparation. The sections were evaluated under the polarizing light microscopy for remineralizing efficacy. The lesion depth was measured and tabulated to be sent for statistical analysis. RESULTS: The mean demineralization value for nonfluoride and herbal-based dentifrice groups were 7.8730 µm and 28.3174 µm, respectively. Hence, it can be inferred that since lesion depth measured was lesser in nonfluoride than herbal-based dentifrice, remineralization has occurred in the nonfluoride-based dentifrice group. CONCLUSION: Nonfluoride-based dentifrice showed significant results in remineralizing the demineralized lesion, while herbal-based dentifrice showed poor efficiency in remineralizing the demineralized lesion.

In vitro demineralization prevention by fluoride and silver nanoparticles when applied to sound enamel and enamel caries-like lesions of varying severities.

OBJECTIVES: To investigate the effect of fluoride and silver nanoparticles on the prevention of in vitro demineralization of sound enamel and enamel caries-like lesions of varying severities. METHODS: Caries-like lesions of different severities (1/6/15 days) were created in bovine enamel specimens. One group remained sound. All specimens were demineralized again using a partially saturated acetic acid solution. Mimicking the intra-oral retention of fluoride and silver in vitro, this solution was supplemented with fluoride (0/1/10 ppm) and/or silver nanoparticles (0/10 ppm) in a factorial design. Changes in lesion depth (ΔL) and integrated mineral loss (ΔΔZ) were evaluated by digital transverse microradiography. Data was analyzed using three-way ANOVA. RESULTS: Lesion severity significantly affected ΔΔZ and ΔL, after no treatment and after the treatment of fluoride and silver independently (p = 0.012 and p = 0.037, respectively). Fluoride and the fluoride × lesion severity interaction were shown to be significant (p < 0.001) on ΔΔZ and ΔL. Silver nanoparticles significantly affected ΔΔZ (p = 0.041), but not ΔL (p = 0.15). The silver nanoparticles × lesion severity interaction was significant for ΔΔZ and ΔL (p = 0.032 and p = 0.024, respectively). No interaction was observed for ΔΔZ and ΔL between fluoride and silver (p = 0.962 and p = 0.971, respectively) as well as lesion severity and the use of fluoride and silver combined (p = 0.722 and p = 0.158, respectively). CONCLUSION: Fluoride and silver nanoparticles had a significant effect on the prevention of in vitro demineralization of sound enamel and enamel caries-like lesions of varying severities. CLINICAL SIGNIFICANCE: Fluoride and silver nanoparticles may potentially allow for more tailored caries prevention.

Effect of Aerva sanguinolenta (Lal bishalyakarani) plant extract on biofilm-induced human enamel demineralization: An in vitro study.

BACKGROUND: Controlling cariogenic biofilm formation by plant extracts could add to preventive strategies to dental caries. OBJECTIVE: To evaluate in vitro the role of Aerva Sanguinolenta ethanolic extract on biofilm-induced microbial human enamel demineralization. METHODOLOGY: The prepared enamel sections of study group (SG), positive control group (PCG), and negative control group (NCG) were immersed in 2 ml of 0.2% ethanolic extract of A. sanguinolenta, 0.12% chlorhexidine, and distilled water, respectively, for 2 min before subjecting to closed batch culture technique utilizing mono- or dual-species culture media of Streptococcus mutans and Lactobacillus acidophilus. Quantification of biofilm and demineralization of enamel was performed by crystal violet (CV) assay and scanning electron microscope (SEM) attached to energy-dispersive X-ray analysis, respectively. STATISTICAL ANALYSIS: Two-way ANOVA and Tukey's test were used for analysis. RESULTS: CV assay of biofilm recorded the highest and lowest optical absorbance value in NC3 (2.728660) and PC3 (0.364200), respectively. Thus, biofilm formation is highest in NCG and lowest among PCG. Surface roughness and porosity in enamel are greatest among NCG and lowest among SG as evident by SEM. Wt% of calcium (S3 47.7170) and phosphorus ion (S3 22.7330) was highest in SG, closely resembling that of B enamel (Ca = 41.9530, P = 19.6650). Wt% of oxygen is lowest in SG (S3 28.8920) and resembles baseline O2 (37.4950). Thus, the amount of biofilm formation is moderate and amount of demineralization of enamel is least among SGs. CONCLUSION: Enamel exposed to 2 ml of 0.2% solution of A. sanguinolenta for 2 min could fairly inhibit formation of biofilm and positively inhibit underlying demineralization in cariogenic environment.

Anti-caries effect of fluoridated milk-based drink consumed by older adults on an in vitro root caries experimental model.

OBJECTIVE: This study aimed to evaluate the anti-caries effect of a fluoridated milk-based drink on a root caries model by assessing mineral loss and both biofilm microbial viability and acidogenicity under increasing concentrations of fluoride supplementation. DESIGN: Streptococcus mutans UA159 biofilms were grown on root dentin slabs for five days. The slabs were randomly assigned to following groups: milk-based drink (G1) and milk-based drink supplemented with 5-ppm NaF (G2), 10 ppm NaF (G3), and 20 ppm NaF (G4). A 10% sucrose and 0.9% NaCl solution were used as positive and negative-caries controls, respectively. Slabs/biofilms were exposed to the different treatments 3 times/day for 5 min. To estimate biofilm acidogenicity, the pH of the spent media was serially measured to calculate the area above the curve. Viable bacteria and dentin demineralization were assessed after the experimental phase. Results were compared using ANOVA followed by the Tukey test. RESULTS: G1 exhibited slightly lower acidogenicity than the positive caries control group (p < 0.05). G2, G3, and G4 induced lower acidogenicity than 10% sucrose and the non-supplemented milk-based drink. The lowest acidogenicity was found in G4 (p < 0.05). Fluoride-supplemented milk-based drinks (G2, G3, and G4) resulted in lower bacterial counts (p < 0.05) and induced lower demineralization (p < 0.05) than the positive caries control and non-supplemented milk-beverage (G1). There was a dose-dependent inhibition of demineralization with fluoride-supplemented milk-based drinks. CONCLUSIONS: Fluoride supplementation of a milk-based drink for older adults may reduce its cariogenicity in root dentin.

Combined effects of a topical fluoride treatment and 445 nm laser irradiation of enamel against a demineralization challenge: A light and electron microscopic ex vivo study.

This study investigated the caries-preventive effect of 445 nm laser radiation in combination with fluoride on the prevention of white spot lesions. Previously, several studies have indicated the ability of 488 nm argon ion laser irradiation to reduce early enamel demineralization. A diode laser (445 nm) could be an alternative technology for possible caries-preventive potential. Each sample of a group of seventeen caries-free bovine teeth was treated in four different ways on four different zones of the labial surface: control/no treatment (C), laser irradiation only (L) (0.3 W, 60 s and applied dose of 90 J/cm2), amine fluoride application only (10,000 ppm and pH 3.9) (F), and amine fluoride application followed by laser irradiation (FL). After treatment, the teeth were subjected to a demineralization solution (pH 4.3 for 48 h at 37 °C) to induce subsurface lesions. After sectioning, the teeth were examined by light microscopy. Three teeth were analyzed by scanning electron microscopy (SEM). The depths of the subsurface lesions in the C, L, F, and FL groups were 103.01 (± 13.04), 96.99 (± 14.51), 42.59 (± 17.13), and 24.35 (± 11.38) µm, respectively. The pairwise group comparison showed the following results: p < 0.001 for FL versus C, FL versus L, F versus C, and F versus L, p = 0.019 for FL versus F and p = 0.930 for L versus C. The SEM micrographs support the light-microscopic examination. The results of the current study have shown that using relatively low irradiation settings of 445 nm laser on fluoridated enamel may be effective for prevention of white spot lesions.

Effectiveness of in vitro primary coronal caries prevention with silver diamine fluoride - Chemical vs biofilm models.

OBJECTIVES: The main goal of this study was to investigate the effectiveness of SDF and its individual components, silver (Ag+) and fluoride (F-) ions, in preventing enamel demineralization using biofilm and chemical models. METHODES: Polished human enamel specimens were assigned to five treatment groups (n = 18 per group): SDF (38 %); SDF followed by application of a saturated solution of potassium iodide (SDF + KI); silver nitrate (AgNO3; silver control, 253,900 ppm Ag+); potassium fluoride (KF; fluoride control, 44,800 ppm F); deionized water (DIW). Treatments were applied once to sound enamel. In the biofilm model, specimens were demineralized by aerobic overnight incubation using cariogenic bacteria isolated from human saliva in brain heart infusion supplemented with 0.2 % sucrose for three days. In the chemical model, enamel specimens were immersed in a demineralizing solution containing 0.1 M lactic acid, 4.1 mM CaCl2, 8.0 mM KH2PO4, 0.2 % Carbopol 907, pH adjusted to 5.0 for five days. Vickers surface microhardness was used to determine the extent of enamel demineralization. Data were analyzed using one-way ANOVA. RESULTS: In the chemical model, there was no statistically significant difference between SDF and SDF + KI in preventing coronal caries (p < 0.0001). In the biofilm model, SDF + KI was significantly less effective in preventing demineralization than SDF (p < 0.0001). In both models, SDF and SDF + KI were superior in their ability to prevent caries lesion formation than AgNO3 and DIW. CONCLUSION: KI application after SDF treatment appears to impair SDF's ability to prevent biofilm-mediated but not chemically induced demineralization. CLINICAL SIGNIFICANCE: SDF may be a viable option in preventing primary coronal caries.

Effect of Er,Cr:YSGG Laser on the Prevention of Primary and Permanent Teeth Enamel Demineralization: SEM and EDS Evaluation.

Objective: To evaluate in vitro the effect of the erbium, chromium:yttrium-scandium-gallium-garnet (Er,Cr:YSGG) laser on resistance of primary and permanent human enamel to demineralization using water cooling and fluoride coapplication as variable parameters. Methods: Enamel specimens were prepared from extracted primary and permanent teeth (n = 225 each). The specimens were separated into 15 subgroups (n = 15/group) based on laser application at three different power settings (0.25, 0.50, and 0.75 W), laser application with and without water cooling, and application of acidulated phosphate fluoride (APF) gel before laser treatment. Morphological changes were assessed with scanning electron microscopy (SEM), and the specimens' chemical contents were determined with energy-dispersive X-ray spectroscopy. Results: In both the primary and permanent teeth, the highest Ca and P content was observed in the noncooled 0.75 W laser group (p < 0.05), irrespective of APF pretreatment (p > 0.05). The Ca and P content for the noncooled APF +0.75 W laser group was lower than that for the APF group and the noncooled 0.75 W laser group. For both dentitions, the F mass content for the APF+laser groups was significantly higher than laser-only groups (p < 0.05). Under SEM, both the primary and permanent enamel exhibited cracks, craters, and surface roughness without water cooling, consistent with increased power output. Conclusions: Er,Cr:YSGG laser application at 0.75 W without water cooling increased enamel resistance to demineralization. Compared with topical APF application, Er,Cr:YSGG laser application barely improved enamel resistance against demineralization, and coapplication did not result in a synergistic effect.

Influence of cigarette smoke combined with different toothpastes on enamel erosion.

This in vitro study aimed to evaluate the effect of different toothpastes on dental enamel subjected to an erosive cycle with and without exposure to cigarette smoke. Bovine enamel specimens were randomly allocated into 12 groups (n = 12). For the in vitro simulation of smoking, half the groups underwent an exposure cycle of 20 cigarettes per day for 5 days. Subsequently, all groups were subjected to a 5-day erosion cycle intercalating demineralization (1 min; 1% citric acid; pH = 3.5) and treatment with toothpaste slurries (2 min) of NaF, SnF2, F/Sn/Chitosan, F/CaSiO3/Na3PO4, and F/bioactive glass. The control group was immersed in distilled water. Surface microhardness (SMH) was measured initially, after exposure to smoke, and after the erosive cycle, and %SMH was calculated. At the end of the experimental cycle, surface roughness, profilometry, and atomic force microscopy (AFM) were performed. SMH increased after exposure to cigarette smoke (p < 0.05). After the erosive cycle, there were no differences between the presence and absence of cigarette smoke exposure in SMH and roughness (p > 0.05). Besides increasing enamel SMH, cigarette smoke did not prevent enamel loss after the erosion cycle (p < 0.05). In profilometry, roughness and surface loss had the lowest values in the groups treated with SnF2 and F/Sn/Chitosan (p < 0.05). AFM showed lower mineral loss with F/CaSiO3/Na3PO4 and F/Sn/Chitosan. For all groups, except F/CaSiO3/Na3PO4, cigarette smoke resulted in higher enamel wear. F/Sn/Chitosan showed the best results against erosion.

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.

Effect of hydroalcoholic extract of Myracrodruon urundeuva All. and Qualea grandiflora Mart. leaves on the viability and activity of microcosm biofilm and on enamel demineralization.

OBJECTIVES: The aim of this study was to assess the effect of Myracrodruon urundeuva All. and Qualea grandiflora Mart. leaves hydroalcoholic extracts on viability and metabolism of a microcosm biofilm and on enamel demineralization prevention. METHODOLOGY: Microcosm biofilm was produced on bovine enamel using inoculum from pooled human saliva mixed with McBain saliva, under 0.2% sucrose exposure, for 14 days. The biofilm was daily-treated with the extracts for 1 min. At the end, it was analyzed with respect to viability by fluorescence, CFU counting and extracellular polysaccharides (phenol-sulphuric acid colorimetric assay) and lactic acid (enzymatic assay) production. The demineralization was measured by TMR. The data were compared using ANOVA or Kruskal-Wallis (p<0.05). RESULTS: M. urundeuva All. at 100, 10 and 0.1 µg/mL and Q. grandiflora Mart. at 100 and 0.1 µg/mL reduced biofilm viability similarly to positive control (chlorhexidine) and significantly more than the negative-vehicle control (35% ethanol). M. urundeuva at 1000, 100 and 0.1 µg/mL were able to reduce both lactobacilli and mutans streptococci CFU counting, while Q. grandiflora (1000 and 1.0 µg/mL) significantly reduced mutans streptococci CFU counting. On the other hand, the natural extracts were unable to significantly reduce extracellular polysaccharides and lactic acid productions neither the development of enamel carious lesions. CONCLUSIONS: The extracts showed antimicrobial properties on microcosm biofilm, however, they had no effect on biofilm metabolism and caries protection.

Effect of hydroalcoholic extract of Myracrodruon urundeuva All. and Qualea grandiflora Mart. leaves on the viability and activity of microcosm biofilm and on enamel demineralization

Abstract Objectives: The aim of this study was to assess the effect of Myracrodruon urundeuva All. and Qualea grandiflora Mart. leaves hydroalcoholic extracts on viability and metabolism of a microcosm biofilm and on enamel demineralization prevention. Methodology: Microcosm biofilm was produced on bovine enamel using inoculum from pooled human saliva mixed with McBain saliva, under 0.2% sucrose exposure, for 14 days. The biofilm was daily-treated with the extracts for 1 min. At the end, it was analyzed with respect to viability by fluorescence, CFU counting and extracellular polysaccharides (phenol-sulphuric acid colorimetric assay) and lactic acid (enzymatic assay) production. The demineralization was measured by TMR. The data were compared using ANOVA or Kruskal-Wallis (p<0.05). Results: M. urundeuva All. at 100, 10 and 0.1 μg/mL and Q. grandiflora Mart. at 100 and 0.1 μg/mL reduced biofilm viability similarly to positive control (chlorhexidine) and significantly more than the negative-vehicle control (35% ethanol). M. urundeuva at 1000, 100 and 0.1 μg/mL were able to reduce both lactobacilli and mutans streptococci CFU counting, while Q. grandiflora (1000 and 1.0 μg/mL) significantly reduced mutans streptococci CFU counting. On the other hand, the natural extracts were unable to significantly reduce extracellular polysaccharides and lactic acid productions neither the development of enamel carious lesions. Conclusions: The extracts showed antimicrobial properties on microcosm biofilm, however, they had no effect on biofilm metabolism and caries protection.

In situ effect of fluoride toothpaste supplemented with nano-sized sodium trimetaphosphate on enamel demineralization prevention and biofilm composition.

OBJECTIVE: To evaluate the effect of a fluoride toothpaste containing nano-sized sodium trimetaphosphate (TMPnano) on enamel demineralization in situ and composition of the biofilm. DESIGN: This crossover double-blind study consisted of four phases (seven days each) and 12 volunteers who wore oral appliances containing four enamel bovine blocks. The cariogenic challenge was performed by 30% sucrose solution (6x/day). The toothpaste treatments (3x/day) were as follows: no F/TMP/TMPnano (Placebo), 1100 ppm F (1100F), 1100F plus 3% micrometric or nano-sized TMP (1100F/TMP; 1100F/TMPnano). Percentage of surface hardness loss (%SH), and integrated loss of subsurface hardness (ΔKHN), as well as enamel calcium (Ca), phosphorus (P), and fluoride (F) were determined. Moreover, biofilm formed on the blocks were analyzed for F, Ca, P, and insoluble extracellular polysaccharide (EPS) concentrations. Data were analyzed using one-way ANOVA, repeated measures followed by Fisher LSD test (p < 0.001). RESULTS: 1100F/TMPnano promoted the lowest %SH and ΔKHN among all groups (p < 0.001). Regarding the F concentrations in the enamel and in the biofilm, there were no significant differences between 1100 F and 1100 F/TMPnano, but significantly increased enamel Ca concentrations (p < 0.001). 1100F/TMPnano showed lower values of EPS concentration when compared with 1100F (∼80%) (p < 0.001). CONCLUSION: 1100F/TMPnano promoted a greater protective effect against enamel demineralization and significantly affected the composition of biofilm formed in situ when compared to 1100F toothpaste.

Hydroalcoholic extracts of Myracrodruon urundeuva All. and Qualea grandiflora Mart. leaves on Streptococcus mutans biofilm and tooth demineralization.

OBJECTIVES: This study evaluated the effect of the hydroalcoholic extracts of Myracrodruon urundeuva All. and Qualea grandiflora Mart. leaves (alone or combined) on the viability of Streptococcus mutans biofilm and on the prevention of enamel demineralization. METHODS: Strain of S. mutans (ATCC 21175) was reactivated in BHI broth. Minimum inhibitory concentration, minimum bactericidal concentration, minimum inhibition biofilm concentration and minimum eradication biofilm concentration were determined in order to choose the concentrations to be tested under biofilm model. S. mutans biofilm (5 × 105 CFU/ml) was produced on bovine enamel, using McBain saliva under 0.2% sucrose exposure, for 3 days. The biofilm was daily treated with the extracts for 1 min. The biofilm viability was tested by fluorescence and the enamel demineralization was measured using TMR. RESULTS: Myracrodruon urundeuva All. (Isolated or combined) at the concentrationsc ≥0.625 mg/ml was able to reduce bacteria viability, while Qualea Grandflora Mart. alone had antimicrobial effect at 5 mg/ml only (p < 0.05). On the other hand, none of the extracts were able to reduce enamel demineralization. CONCLUSIONS: The hydroalcoholic extracts of Myracrodruon urundeuva All. and Qualea grandiflora Mart. leaves (isolated or combined) have antimicrobial action; however, they do not prevent enamel caries under S. mutans biofilm model.

Remineralizing effect of a fluoridated gel containing sodium hexametaphosphate: An in vitro study.

OBJECTIVES: To evaluate in vitro the effect of neutral pH topical gels with reduced fluoride concentration (F), supplemented or not with sodium hexametaphosphate (HMP) on the remineralization of dental enamel, using a pH-cycling model. Materials and methods Bovine enamel blocks with caries-like lesions were randomly treated with five gels (n = 24/group): without F/HMP (Placebo); 4500 ppm F (4500F), 4500F plus 9% HMP (4500F + HMP); 9000 ppm F (9000F); and 12,300 ppm F (Acid gel). After pH-cycling, the percentage of surface hardness recovery (%SHR), integrated loss of subsurface hardness (ΔKHN), and concentrations of loosely- (CaF2) and firmly-bound (FA) fluoride formed and retained in/on enamel were determined. The results were analyzed by ANOVA followed by the Student-Newman-Keuls test (p < 0.001). RESULTS: The 4500F + HMP gel promoted the highest %SHR among all groups; the lowest ΔKHN was achieved by 4500F + HMP and Acid gel, without significant differences between these. The Acid gel group presented the highest CaF2 and FA formed and retained on/in enamel (p < 0.001). CONCLUSION: Based on the present results, the addition of 9% sodium hexametaphosphate to a gel with reduced fluoride concentration (4500F) was able to significantly enhance the remineralization of artificial carious lesions in vitro when compared to 4500F, reaching protective levels similar to an acidic formulation with ∼3-fold higher fluoride concentration.

A comparative evaluation of APF gel, CPP/ACP paste alone and in combination with carbon dioxide laser on human enamel resistance to acid solubility using atomic absorption spectrometry: an in-vitro study.

BACKGROUND: The aim of this study was to compare the effects of acidulated phosphate fluoride (APF) gel, calcium phosphopeptide-amorphous calcium phosphate (CPP/ACP) paste alone and in combination with CO2 laser on the resistance of enamel to acid solubility. METHODS: Ninety enamel sections were obtained from 15 extracted teeth and were randomly assigned to six groups: 1) control group; 2) APF group; 3) CPP-ACP group; 4) CO2 laser group; 5) APF + CO2 group; and 6) CPP-ACP + CO2 group. The specimens were individually demineralized in 0.1 M lactic acid solution with adjusted pH of 4.8 for 24h at 37 ºC. The acid solubility was determined using atomic absorption spectrometry. Statistical analysis was done using one-way ANOVA and Tukey-Kramer post hoc test (P<0.05). RESULTS: The average extent of calcium ion released (ppm) was estimated as follow: group 1: 6.974±1.757, group 2: 5.363±1.383, group 3: 6.962±1.489, group 4: 6.890±1.560, group 5: 4.803±1.080 and group 6: 6.789±1.218. Based on the between-group comparison results, group 2 and group 5 showed significant differences with the other groups. CONCLUSIONS: Under the studied conditions, only, the APF group alone and in combination with CO2 laser could decrease enamel acid solubility.