Enamel remineralisation prospect of Moringa Oleifera hydrogel, eggshell hydrogel versus sodium fluoride varnish on artificially demineralised primary teeth: in vitro study.

PURPOSE: The purpose of the present in vitro study is to investigate and compare the remineralising potential of Moringa Oleifera extract, eggshell, and sodium fluoride varnish on microhardness of artificially demineralised enamel of primary teeth with biomimetic minimally invasive approach following the world paradigm shift towards natural products in paediatric dentistry. MATERIAL AND METHODS: Sample size included 44 primary molars. The mineral content and surface microhardness of all specimens were initially assessed using energy dispersive x-ray examination (EDX) and Vickers microhardness. The specimens were artificially demineralised for 96 h at a temperature of 37°C and then reassessed directly after demineralisation. The demineralised enamel specimens were randomly divided into four groups according to the remineralisation regimen utilised. Group 1: Artificial saliva (control); Group 2: Sodium fluoride varnish; Group 3: Eggshell hydrogel; and Group 4: Moringa Oleifera hydrogel. The specimens were stored for 8 days and then subsequently evaluated using EDX and microhardness assessment by Vickers microhardness test and scanning electron microscope (SEM).  Results: Regarding the microhardness test, there was a significant difference between the Moringa Oleifera group and Eggshell group compared to fluoride varnish (p < 0.05). Regarding EDX analysis, there was a statistically significant difference (p < 0.05) between Moringa Oleifera group and Eggshell group compared to fluoride varnish as the highest values were for Moringa Oleifera and Eggshell. On the other hand, there was no statistically significant difference (p > 0.05) between Moringa Oleifera and Eggshell in both the measurements. CONCLUSION: Moringa Oleifera and Eggshell might be considered as a biomimetic natural material capable of guiding enamel tissue remineralisation in early carious lesion of primary teeth. CLINICAL RELEVANCE: This research demonstrated the capability for early enamel caries to be remineralised using novel materials with a naturally counterpart implicated in biomineralisation as proved to be more effective than traditionally used fluoride varnish in primary teeth.

Xylitol associated or not with fluoride: Is the action the same on de- and remineralization?

OBJECTIVES: This study evaluated the effect of xylitol combined or not with fluoride (F) on reduction of demineralization and increase of remineralization of shallow and deep artificial enamel lesions. METHODS: Bovine enamel samples were allocated to the following solutions groups: no xylitol (negative control), 5% xylitol, 10% xylitol, 20% xylitol, 500 ppm F (as NaF), 5% xylitol+F, 10% xylitol+F or 20% xylitol+F (n = 12-15). For the demin study, a pH-cycling model (demineralization-6 h, pH 4.7/remineralization 18 h, pH 7.0) was employed for 7 days. Treatments were applied 2 × 1 min. In the remin study, specimens were pre-demineralized for 2, 5 or 10 days. Afterwards, a pH-cycling protocol was conducted (2 h demineralizing and 22 h remineralizing solution/day for 8 days) and the same treatments were done. The response variables were percentage surface hardness loss (%SHL) and transverse microradiography. Data were analyzed by RM ANOVA/Tukey or Kruskal-Wallis/Dunn (p < 0.05) RESULTS: F and Xylitol combined with F reduced the %SHL (23-30%) compared to the negative control (61.5%). The integrated mineral loss and the lesion depth were not reduced by any treatment. Surface hardness recovery was seen only for shallow lesions in case of 20% xylitol+F compared to negative control. No lesion depth recovery, but significant mineral recovery was seen for F (2-days and 10-days lesion). CONCLUSIONS: All concentrations of xylitol+F reduced enamel surface demineralization, while only 20% xylitol+F improved surface remineralization of shallow lesions in vitro. CLINICAL SIGNIFICANCE: Our results suggest that while F or any concentration of xylitol + F reduces surface demineralization, only 20% xylitol+F improves surface remineralization of shallow lesions in vitro. Therefore, xylitol may be added into oral products, combined to F, to control dental caries.

Estimation of the Efficacy of Remineralizing Agents on the Microhardness of Deciduous Teeth Demineralized Using Pediatric Formulations.

AIM: This study aimed to evaluate the demineralizing effect of commonly used pediatric syrup formulations on primary teeth and the efficacy of two readily available remineralizing agents in treating this effect. MATERIALS AND METHODS: Ninety primary teeth were used for sample preparation and divided into three groups: antibiotic syrup (group A), cough syrup (group B), and control (group C) groups. These groups were further categorized into intragroups according to the treatment with remineralizing agents: groups A1, B1, and C1 received GC Tooth Mousse (casein phosphopeptide-amorphous calcium phosphate, CPP-ACP paste) and groups A2, B2, and C2 received Clinpro Tooth Crème. The samples were subjected to a series of demineralization cycles for 14 days, and remineralization cycles until 30 days were performed using two remineralizing agents, that is, GC Tooth Mousse (CPP-ACP paste) and Clinpro Tooth Crème and were evaluated using Vicker's microhardness test. RESULTS: Antibiotic syrup (group A) and cough syrup (group B) showed a significant decrease in surface microhardness compared with control (group C). All intragroups showed an increase in surface microhardness after treatment with remineralizing agents, which was significantly higher in intragroups A1, B1, and C1 treated with GC Tooth Mousse (CPP-ACP paste). CONCLUSIONS: Oral liquid medications showed definite demineralization potential. CPP-ACP paste was found to be better than Clinpro Tooth Crème for demineralized teeth. CLINICAL SIGNIFICANCE: The use of over-the-counter drugs has increased among the average Indian population, especially for the treatment of fever, cold, and cough. Unwise use of medications by the present population without proper medical guidance will lead to irreparable changes in future generations.

Evaluation of the effects of different mouthrinses on dental remineralization.

BACKGROUND: Dental caries occurs with the release of organic acids from the fermentable carbohydrates metabolized by cariogenic microorganisms. Microbial, genetic, immunological, behavioral, and environmental factors play a role in the development and severity of dental caries. OBJECTIVES: The aim of the present study was to investigate the possible effects of different mouthwash solutions on dental remineralization. MATERIAL AND METHODS: This in vitro study compared the remineralization capacity of different mouthwash solutions applied topically to the enamel surface. A total of 50 tooth specimens were prepared from the buccal and lingual halves, with 10 teeth in each group: G1 (control); G2 (Listerine®); G3 (Sensodyne®); G4 (Oral B® Pro-Expert); and G5 (DentaSave® Zinc). Remineralization capacity was evaluated in all groups. The one-way analysis of variance (ANOVA) and the paired samples t test were used for statistical analysis, with a p-value <0.05 considered significant. RESULTS: There were significant differences in the calcium (Ca)/phosphorus (P) atomic percentage (at%) ratio between the demineralized and remineralized dentin (p = 0.001), and between the demineralized and remineralized enamel (p = 0.006). Similarly, there were significant differences in the at% of P (p = 0.017) and zinc (Zn) (p = 0.010) between the demineralized and remineralized dentin. There was a significant difference in the at% of P (p = 0.030) between the demineralized and remineralized enamel. The Zn at% in enamel was significantly higher after remineralization in G5 as compared to the control group (p < 0.05). The images of the demineralized enamel showed the usual keyhole prism appearance, with intact prism sheaths and negligible inter-prism porosity. CONCLUSIONS: The scanning electron microscopy (SEM) and energy-dispersive X-ray spectroscopy (EDS) findings seem to confirm the effectiveness of DentaSave Zinc for the remineralization of enamel lesions.

In vitro remineralization assessment of enamel subsurface lesions using different percentages of surface reaction-type pre-reacted glass-ionomer containing gum-based material.

OBJECTIVE: To identify the remineralization activity of enamel subsurface lesions using different percentages of surface pre-reacted glass-ionomer (S-PRG) filler containing gum-base material. METHODS: Gum extracts from gum-base materials containing 0wt%, 5wt%, and 10wt% S-PRG filler were prepared as GE0, GE5, and GE10, respectively. A total of 50 bovine enamel specimens were used, and the polished enamel surface of a 3 × 3 mm2 window area was exposed. The specimens were then subjected to a demineralization solution for seven days to create a subsurface enamel lesion. Remineralization was then conducted for seven days using a protocol whereby the specimens were immersed three times a day in prepared gum extracts (0wt%, 5wt%, and 10wt%) and artificial saliva of pH 7 (Control) for 20 min at 37 °C. Thereafter, remineralization assessment was performed by using Swept Source Optical Coherence Tomography (SS-OCT) and micro-computed tomography (µCT). Surface morphology and elemental analysis were conducted by scanning electron microscopy (SEM) and energy-dispersive X-ray spectrometry (EDS). RESULTS: The depths of the demineralized lesions in the GE5 and GE10 groups were significantly lower than those of the Control and the GE0 groups. SEM observations of the enamel surface morphology of the GE5 and GE10 groups indicated remineralization with S-PRG filler-related elements present. CONCLUSION: The GE5 and GE10 S-PRG filler containing gum-base materials showed significantly improved surface remineralization and reduced demineralization of the enamel lesions. EDS analysis suggested that the released ions from the S-PRG filler might be responsible for surface remineralization. CLINICAL SIGNIFICANCE: The S-PRG filler containing gum-base material may have a significant remineralization effect and improve the surface morphology of enamel subsurface lesions.

Effect of fluoride gels with nano-sized sodium trimetaphosphate on the in vitro remineralization of caries lesions.

OBJECTIVE: To evaluate the effects of fluoride (F) gels supplemented with micrometric or nano-sized sodium trimetaphosphate (TMPmicro and TMPnano, respectively) on the in vitro remineralization of caries-like lesions. METHODOLOGY: Bovine enamel subsurface lesions (n=168) were selected according to their surface hardness (SH) and randomly divided into seven groups (n=24/group): Placebo (without F/TMP), 4,500 ppm F (4500F), 4500F + 2.5% TMPnano (2.5% Nano), 4500F + 5% TMPnano (5% Nano), 4500F + 5% TMPmicro (5% Micro), 9,000 ppm F (9000F), and 12,300 ppm F (Acid gel). The gels were applied in a thin layer for one minute. Half of the blocks were subjected to pH cycling for six days, whereas the remaining specimens were used for loosely- (calcium fluoride; CaF2) and firmly-bound (fluorapatite; FA) fluoride analysis. The percentage of surface hardness recovery (%SHR), area of subsurface lesion (ΔKHN), CaF2, FA, calcium (Ca), and phosphorus (P) on/in enamel were determined. Data (log10-transformed) were subjected to ANOVA and the Student-Newman-Keuls' test (p<0.05). RESULTS: We observed a dose-response relation between F concentrations in the gels without TMP for %SHR and ΔKHN. The 2.5% Nano and 5% Micro reached similar %SHR when compared with 9000F and Acid gels. For ΔKHN, Placebo and 5% Nano gels had the highest values, and 5% Micro, 2.5% Nano, 9000F, and Acid gels, the lowest. All groups had similar retained CaF2 values, except for Placebo and Acid gel. We verified observed an increase in Ca concentrations in nano-sized TMP groups. Regarding P, TMP groups showed similar formation and retention to 9000F and Acid. CONCLUSION: Adding 2.5% nano-sized or 5% micrometric TMP to low-fluoride gels lead to enhanced in vitro remineralization of artificial caries lesions.

In vitro effect of low-fluoride toothpaste supplemented with sodium trimetaphosphate, xylitol, and erythritol on enamel demineralization.

OBJECTIVE: Regular use of toothpaste with fluoride (F) concentrations of ≥ 1000 ppm has been shown to contribute to reducing caries increment. However, when used by children during the period of dental development, it can lead to dental fluorosis. In this study, we aimed to evaluate the in vitro effect of a toothpaste formulation with reduced fluoride (F) concentration (200 ppm) supplemented with sodium trimetaphosphate (TMP: 0.2%), Xylitol (X:16%), and Erythritol (E: 4%) on dental enamel demineralization. METHODOLOGY: Bovine enamel blocks were selected according to initial surface hardness (SHi) and then divided into seven experimental toothpaste groups (n=12). These groups included 1) no F-TMP-X-E (Placebo); 2) 16% Xylitol and 4% Erythritol (X-E); 3) 16% Xylitol, 4% Erythritol and 0.2%TMP (X-E-TMP); 4) 200 ppm F (no X-E-TMP: (200F)); 5) 200 ppm F and 0.2% TMP (200F-TMP); 200 ppm F, 16% Xylitol, 4% Erythritol, and 0.2% TMP (200F-X-E-TMP); and 7) 1,100 ppm F (1100F). Blocks were individually treated 2×/day with slurries of toothpastes and subjected to a pH cycling regimen for five days (DES: 6 hours and RE: 18 hours). Then, the percentage of surface hardness loss (%SH), integrated loss of subsurface hardness (ΔKHN), fluoride (F), calcium (Ca), and phosphorus (P) in enamel were determined. The data were analyzed by ANOVA (1-criterion) and the Student-Newman-Keuls test (p<0.001). RESULTS: We found that the 200F-X-E-TMP treatment reduced %SH by 43% compared to the 1100F treatments (p<0.001). The ΔKHN was ~ 65% higher with 200F-X-E-TMP compared to 1100F (p<0.001). The highest concentration of F in enamel was observed on the 1100F treatment (p<0.001). The 200F-X-E-TMP treatment promote higher increase of Ca and P concentration in the enamel (p<0.001). CONCLUSION: The association of 200F-X-E-TMP led to a significant increase of the protective effect on enamel demineralization compared to the 1100F toothpaste.

Effects of Infantile Hypophosphatasia on Human Dental Tissue.

Hypophosphatasia (HPP) is an inherited, systemic disorder, caused by loss-of-function variants of the ALPL gene encoding the enzyme tissue non-specific alkaline phosphatase (TNSALP). HPP is characterized by low serum TNSALP concentrations associated with defective bone mineralization and increased fracture risk. Dental manifestations have been reported as the exclusive feature (odontohypophosphatasia) and in combination with skeletal complications. Enzyme replacement therapy (asfotase alfa) has been shown to improve respiratory insufficiency and skeletal complications in HPP patients, while its effects on dental status have been understudied to date. In this study, quantitative backscattered electron imaging (qBEI) and histological analysis were performed on teeth from two patients with infantile HPP before and during asfotase alfa treatment and compared to matched healthy control teeth. qBEI and histological methods revealed varying mineralization patterns in cementum and dentin with lower mineralization in HPP. Furthermore, a significantly higher repair cementum thickness was observed in HPP compared to control teeth. Comparison before and during treatment showed minor improvements in mineralization and histological parameters in the patient when normalized to matched control teeth. HPP induces heterogeneous effects on mineralization and morphology of the dental status. Short treatment with asfotase alfa slightly affects mineralization in cementum and dentin. Despite HPP being a rare disease, its mild form occurs at higher prevalence. This study is of high clinical relevance as it expands our knowledge of HPP and dental involvement. Furthermore, it contributes to the understanding of dental tissue treatment, which has hardly been studied so far.

Inhibition of tooth demineralization caused by Streptococcus mutans biofilm via antimicrobial treatment using hydrogen peroxide photolysis.

OBJECTIVES: An antimicrobial technique utilizing hydroxyl radicals generated by the photolysis of 3% H2O2 has been developed recently. The present study aimed to evaluate the effect of H2O2 photolysis treatment on tooth demineralization caused by Streptococcus mutans biofilm. MATERIALS AND METHODS: To induce tooth demineralization, S. mutans biofilm was allowed to form on the maxillary first molars collected from Wistar rats via 24-h culturing. The samples were immersed in 3% H2O2 and irradiated with 365-nm LED (H2O2 photolysis treatment). Viable bacterial counts in the biofilm were evaluated immediately after treatment and after an additional 30-h culturing by colony counting. The acidogenicity of the biofilm, re-established 30 h after treatment, was assessed by measuring the pH. The effect of H2O2 photolysis treatment on tooth demineralization was assessed by measuring the depth of the radiolucent layer in micro-CT images. RESULTS: H2O2 photolysis significantly reduced viable bacterial counts in the biofilm to 3.7 log colony forming units (CFU)/sample, while the untreated group had 7.9 log CFU/sample. The pH of the biofilm re-established after treatment (6.6) was higher than that of the untreated group (5.3). In line with the pH measurement, the treatment group had a significantly lower depth of radiolucent layer in dentin than the untreated group. CONCLUSIONS: H2O2 photolysis treatment was effective not only in killing the biofilm-forming S. mutans but also in lowering the acidogenicity of the biofilm. Thus, this technique could inhibit tooth demineralization. CLINICAL RELEVANCE: H2O2 photolysis can be applicable as a new dental caries treatment.

The effect of two calcium phosphate-containing agents on the enamel resistance of permanent molars to demineralization: An experimental study.

BACKGROUND: The main purpose of this experimental study was to determine the in vitro effects of two calcium phosphate-containing agents (Remin Pro® and GC Tooth mousse™) on the enamel resistance of permanent molars to demineralization. METHODS: Fifty extracted human third molars were randomly divided into four groups; that is the control group and three case groups treated with Remin Pro®, GC Tooth mousse™, and sodium fluoride gel. The three case groups were treated with 0.25 ml of the paste associated with each experimental group for 5 min, kept in fluoride-free artificial saliva, and incubated at 37°C for 28 days. After the treatment regimen, 10 samples of each case group were subjected to demineralization using an acetic acid-containing solution, and remineralization using a remineralizing solution. The morphology of enamel was observed via scanning electron microscopy and their enamel calcium/phosphorus (Ca/P) ratios were measured before/after the demineralization cycle with energy-dispersive X-ray analysis. Data were statistically analyzed using one-way analysis of variance and post hoc Tukey tests. RESULTS: The enamel Ca/P ratios in the case study groups were significantly higher than that of the control group before/after the demineralization regimen (p < .0001). However, the ratios were not significantly different between the case study groups after the treatment regimen and demineralization cycle (p > .05). CONCLUSION: The outcomes of the current study indicated that all three agents seemed to increase the enamel resistance of permanent molar teeth to demineralization.

Evaluation of bleaching agent effects on color and microhardness change of silver diamine fluoride-treated demineralized primary tooth enamel: An in vitro study.

BACKGROUND: The present study aimed to assess the impact of application of fluoridated- 10% carbamide peroxide (CP) with or without potassium iodide (KI) on silver diamine fluoride (SDF)-treated enamel surface in the primary teeth. METHODS: After stained-remineralized caries lesions (s-RCLs) creation, 96 teeth were randomly allocated to four experimental groups: Group 1:SDF-treated enamel followed by 8-h/day application of 10% CP for 2 weeks; Group 2: SDF-treated enamel followed by 15-min/day application of 10% CP for 3 weeks; Group 3: SDF + KI-treated enamel followed by 8-h/day application of 10% CP for 2 weeks; and Group 4: SDF + KI-treated enamel followed by 15-min/day application of 10% CP for 3 weeks. Enamel microhardness (EMH) test (n = 12) and spectrophotometric color assessment (n = 12) was performed at four stages: baseline (intact enamel), demineralized enamel, aged remineralized-stained enamel, and after final intervention. Sixteen samples were used for SEM evaluation. Data were analyzed with the paired t-test, one-way ANOVA, and Tukey's post-hoc test (p < 0.05). RESULTS: EMH values in all groups showed significant decrease after demineralization (all, p < 0.00001). All samples showed complete recovery of EMH values (%REMH) after SDF application compared to demineralization (%REMHSDF) (p = 0.971). Bleaching caused a slight decrease in %REMH for all groups. However, the differences were not statistically significant (p = 0.979). SEM findings revealed no changes in enamel porosity after bleaching. Bleaching application ameliorated the discoloration in all groups (all, p < 0.00001). All samples in Groups 2 and 4 had significantly lighter color after 21 days as compared to 14-day exposure to the bleaching material (both, p < 0.00001). CONCLUSIONS: SDF application on demineralized primary tooth enamel completely recovered enamel microhardness. 10% carbamide peroxide effectively bleached SDF stain without causing significant decrease in EMH values. Color improvement was more evident with the use of KI immediately after SDF application. Both 15-min and 8-h application of fluoridated CP resulted in statistically similar color enhancement in primary teeth.

Effect of high-fluoride dentifrice on root dentine and bacterial composition in a multispecies biofilm model.

The present study evaluated the effect of high-fluoride dentifrice on dentine demineralization and bacterial composition in a multispecies biofilm model in vitro. A seven-organism bacterial consortium was grown on bovine dentine discs in a high-throughput active attachment model. The biofilms were submitted twice per day to the following dentifrices treatments: 5,000 ppm F, 1,100 ppm F, with placebo as a negative control. After 5 days of biofilm growth, dentine samples were assessed by transversal microradiography, the biofilm was collected for bacterial counts and the pH of the media was determined. Lower integrated mineral loss values were observed when 5,000 ppm F-treatment was used compared to the other treatments. Overall microbiological counts decreased with increasing F-concentration as well the pH of the media throughout the experiment. The 5,000 ppm F-treatment caused a shift in microbial composition and reduced dentine demineralization in the in-vitro experimental model.

Effectiveness of CPP-ACP and fluoride products in tooth remineralization.

OBJECTIVES: To compare the effectiveness of the casein phosphopeptide-stabilized amorphous calcium phosphate (CPP-ACP) in the in situ remineralization of enamel exposed with two different degrees of preformed enamel lesions. METHODS: One hundred and sixty 3 × 3 × 2 mm human enamel slabs were demineralized and divided into two subgroups according to the baseline surface hardness (SH = B1≤150 and B2 >150). During each of four 10-day experimental periods, 10 participants wore intra-oral removable acrylic palatal expanders with four human enamel slabs with preformed lesions (B1 and B2): CO1 and CO2-Control: silica dentifrice without fluoride; MP1 and MP2: MI Paste; MPP1 and MPP2: MI Paste Plus; and FD1 and FD2: Fluoride dentifrice. The Knoop hardness test (50/15s Micromet 2001, Buehler) was performed after demineralization (B1 and B2) and after treatment (T1 and T2). RESULTS: SH was higher in all treatment groups, when compared with the controls, except for CO2 (Mann-Whitney Wilcoxon Test; p < 0.05). The %SH was similar between MPP2 and FD2 and between MPP2 and MP2; however, FD2 and MPP2 products were more effective in microhardness recovery. In B1, all treatment groups were similar. CONCLUSION: MPP and FD are more effective in preventing demineralization in enamel subsurface lesions.

The Effect of Various Fluoride Products on Dentine Lesions during pH-Cycling.

This study compared the effect of topically applied fluoride products on dentine lesions in an in vitro experiment. Demineralized bovine dentine specimens were treated once with either SDF solution (35,400 ppm F), NaF varnish (22,600 ppm F), TiF4 solution (9,200 ppm F), SnF2 gel (1,000 ppm F), no treatment (control), or preserved as baseline lesions. After the application and subsequent removal of the fluoride products, the specimens were subjected to pH-cycling. Calcium loss and uptake in the de- and remineralization buffers were assessed daily. Fluoride release into the buffers was analyzed on days 1, 2, 3, 5, 8, and 13. After the pH-cycling period, mineral distribution throughout the lesion depth was analyzed using transversal microradiography (TMR). X-ray energy-dispersive spectroscopy (EDS) examined the deposition of silver, titanium, and tin after application of SDF, TiF4, and SnF2, respectively. Overall, calcium loss and uptake analysis in the de- and remineralization buffers revealed that the SDF product was the most effective in inhibiting lesion progression, followed by the TiF4, NaF, and SnF2 products. Fluoride analysis disclosed a steep reduction of the amount of fluoride released into de- and remineralization buffers with time. The fluoride effects on de- and remineralization continued beyond the days that fluoride was released into the buffers. TMR analysis showed significant remineralization in the outer zone of the dentine lesions for all fluoride products, with SDF giving hypermineralization in this zone. In the inner zone, lesions developed in all fluoride groups, with the smallest in the SDF group. EDS showed silver and titanium deposition in depth up to 85 µm and 8 µm, respectively, while no tin deposition was observed. The silver in the dentine lesions did not contribute significantly to the density of the TMR profiles in the SDF group. In conclusion, all topical fluoride products protected the dentine lesions against lesion progression, but at different degrees. SDF showed a superior effect in protection against further demineralization and enhancement of remineralization. This was probably attributed to its fluoride concentration that was the highest among the fluoride products.

Validation of a cariogenic biofilm model by evaluating the effect of fluoride on enamel demineralization.

In vitro biofilm models have been extensively used, but only few of the models available to date had been validated in terms of the dose-response effect of anti-caries and/or antimicrobial substances. Additionally, none of the validated models allow the use of microliter volumes of the treatment solutions, needed mainly to test (screen) novel but expensive substances under development. This study aimed at modifying an in vitro cariogenic Streptococcus mutans biofilm model and validating it by assessing the dose-response effect of fluoride on enamel demineralization. S. mutans cariogenic biofilms were developed on saliva-coated enamel slabs previously bonded to acrylic holders fixed to a lid of a culture plate. Biofilms were incubated 8 h/day in culture medium supplemented with 1% sucrose and then overnight in culture medium with glucose 0.1 mM. Biofilms were also treated 2×/day with 2.0 mL of solutions containing 0, 125, 275 and 1250 µg F/mL (n = 10/group). The replaced culture medium was used to: determine the biofilm acidogenicity; estimate the demineralization of enamel; and monitor the fluoride concentration. At 144 h, biofilms were collected for fluoride concentration analyses, and the fluoride uptake by enamel was determined in each slab. The model showed a dose-response effect of fluoride (R2 = 0.96, p < 0.001) between enamel demineralization and the fluoride concentration of the treatments. Water-soluble and bound biofilm fluoride concentrations (p < 0.007), as well as the firmly-bound fluoride concentration found in enamel (p < 0.0001), increased in a dose-dependent manner. Our model constitutes a validated approach that would allow the assessment of the anticaries potential of novel biotechnological strategies, as in the case of expensive salivary peptides, because it would allow to test the treatment solutions using smaller volumes.

Remineralization Potential of a Combination of Chitosan with Nanohydroxyapatite and a Self-assembling Peptide with Nanohydroxyapatite: An In Vitro Study.

AIM: This study explores the demineralizing potential of the combination of chitosan with nanohydroxyapatite (n-HA) and self-assembling peptides with n-HA. MATERIALS AND METHODS: A total of 66 first premolar teeth of similar dimensions extracted for orthodontic purposes were collected for this study. These were then demineralized and randomly divided into the following three groups (n = 22): (i) Control group, (ii) n-HA + Chitosan (HAC), and (iii) self-assembling peptide + n-HA (SP-HA). The samples in each group were brushed every 24 hours with the respective agent. The specimens were stored in Fusayama Meyer's artificial saliva at room temperature and the solution was replenished daily. Mineral content (Ca, P) and surface morphology of the specimens was analyzed, using scanning electron microscopy and energy dispersion X-ray spectroscopy (SEM-EDAX), before demineralization, at 15 days of remineralization and 30 days of remineralization. A two-way analysis of variance (ANOVA) test followed by Tukey's honest significant difference (HSD) post hoc analysis was used to compare the mean elemental composition of the different groups (p < 0.05). RESULTS: There was no significant difference in the calcium (Ca) and phosphate (P) weight percentage between the different groups at the baseline and after demineralization. The Ca and P weight percentages of all three groups after remineralization for 15 and 30 days showed no significant difference from the baseline or after demineralization. The surface morphology after 15 days of remineralization therapy showed decreased surface porosity and increased mineral deposition in the HAC group than the HP-SA group. Surface morphology after 30 days of remineralization showed a more homogenous and smoother surface in the HAC group than the HP-SA group. CONCLUSION: From the results of this study, it can be concluded that the combination of chitosan with n-HA and self-assembling peptides with n-HA can be considered effective demineralizing agents. CLINICAL SIGNIFICANCE: Considering the non-invasive nature of remineralization therapy understanding the effectiveness of different agents is of utmost importance. The demineralizing properties of chitosan, n-HA and self-assembling peptides make their combinations ideal for studying their effectiveness in treating white spot lesions.

Effect of ion-releasing filler-containing gel application on dentin remineralization using optical coherent tomography.

PURPOSE: To examine the effects of an ion-releasing filler-containing gel on the remineralization of dentin using optical coherence tomography (OCT). METHODS: Dentin slabs of bovine teeth were sliced and shaped into a rectangular form. Specimens were treated with undersaturated 0.1 M lactic acid buffer solution (pH 4.75) for 10 minutes and then placed in artificial saliva (pH 7.0). This procedure was repeated three times a day for 28 days. The dentin remineralization effects of a fluoride/S-PRG filler-containing gel (PRG) and a 38% SDF solution (SDF) on dentin slabs of bovine teeth were compared. After treatment, the dentin slabs were immersed in a 0.1 M lactic acid buffer solution and then placed in artificial saliva. This procedure was repeated three times a day for 28 days. OCT imaging was conducted on the selected location of the dentin surface. The peak intensity and width at 1/e² were recorded in each of the six areas on the sample and averaged. Each group had a sample size of 10. Knoop hardness number (KHN) measurements and scanning electron microscopy (SEM) observations were also conducted. The data for each group were subjected to a one-way repeated-measures ANOVA and Tukey tests (α= 0.05). The samples were also observed using SEM. RESULTS: The peak signal intensities of SDF and PRG decreased on day 7 and then slightly increased during the experimental period for the one-off application groups and then decreased for frequent-time application groups. Although the width at 1/e² in the untreated specimens decreased over the test period, SDF and PRG for the one-off application groups exhibited an increase in widths on day 7 followed by a slight decrease, whereas it increased for the frequent-time application groups. The average KHN of the dentin samples exhibited the same tendency as the width at 1/e². Closure of the dentin tubules and crystal precipitation were detected on the surface of both SDF and PRG groups. CLINICAL SIGNIFICANCE: S-PRG filler-containing gel might have the ability to prevent dentin demineralization and could be useful for the prevention of hard-to-access lesions. This material achieved remineralization of the demineralized root dentin and had the same remineralization ability as SDF in vitro.

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.

Effects of desensitizer containing fluoroaluminocalciumsilicate glass nanoparticles on remineralization of root dentin subsurface lesions in vitro.

We investigated the remineralization effects of Nanoseal (NS) dentin desensitizer on demineralized root dentin. Baseline lesion specimens prepared from bovine root dentin were immersed in artificial saliva (AS) or deionized water (DW) after treatment with NS or fluoride-free Nanoseal (NS(-)). Treatment and control groups comprised: 1, AS; 2, NS/AS; 3, NS(-)/AS; 4,NS/DW; 5, NS(-)/DW; and 6, baseline demineralization. Integrated mineral loss (IML) and lesion depth (LD) were determined by transverse microradiography. Fluoride concentrations in the immersion solutions were measured. AS, NS/AS and NS(-)/AS showed higher mineral volume % at the surface and lesion body than did other groups. NS/AS showed significantly lower IML than did AS. There was no significant difference in IML between NS/AS and NS(-)/AS. The highest concentration of fluoride was in the NS/AS immersion solution. The findings suggest Nanoseal facilitated remineralization of demineralized root dentin, and fluoride and other ions included may have contributed to this effect.

Effectiveness of a toothpaste and a serum containing calcium silicate on protecting the enamel after interproximal reduction against demineralization.

To evaluate the effectiveness of a calcium silicate/phosphate fluoridated tooth paste and a serum compared with a toothpaste containing hydroxyapatite on protecting the enamel after interproximal reduction against demineralization. 3 sets of eleven incisors were created. The teeth underwent interproximal enamel reduction (IER) of 0.5 mm. Each set was allocated to one of three groups: (1) Brushing without toothpaste (control group); (2) Vitis toothpaste + Remin Pro; (3) Regenerate toothpaste + Regenerate Serum. The agents were applied three times a day and specimens subjected to demineralization cycles for 30 days. The weight percentages of calcium (Ca) and phosphorous (P) were quantified by X-ray microfluorescence spectroscopy. Surface microhardness measurements and electron scanning microscopy (SEM) observations were made. Ca data and the Ca/P ratio were significantly higher in Group 3 than the other groups (p < 0.017), while P was significantly lower in Group 3 (p < 0.017). No significant differences were found between Groups 1 and 2 (p > 0.017). Group 3 showed significantly higher microhardness values (p < 0.05) than Group 1. No significant differences were found for other comparisons between groups (p < 0.05). SEM images showed less demineralization in Group 3. The application of a calcium silicate/phosphate fluoridated tooth paste (Regenerate advance) and a dual serum (Regenerate advance enamel serum) protect the enamel with interproximal reduction against demineralization. Therefore, this treatment could be used to prevent the dissolution of hydroxyapatite after IER.