Comparison of various chitosan-based in situ forming gels with sodium fluoride varnish for enamel biomineralization: an in-vitro pH cycling model.

This study aimed to evaluate chitosan (CS)-based formulations loaded with 5% sodium fluoride (NaF) and/or 10% nanohydroxyapatite (nHA) to remineralize the demineralized primary tooth enamel surface. Ninety enamel blocks were demineralized and were divided into six groups (n = 15): (1) CS-based hydrogel, (2) CS-based hydrogel loaded with NaF, (3) CS-based hydrogel loaded with nHA, (4) CS-based hydrogel loaded with NaF and nHA, (5) 5% NaF varnish, and (6) negative control with no intervention. After intervention, the specimens were pH cycled by 2 h immersion in demineralizing solution and 22 h immersion in remineralizing solution for 8 days. The remineralization effects were evaluated by Vickers microhardness measurements and field emission scanning electron microscopy coupled with energy-dispersive X-ray spectrometry (FESEM-EDS). The best mean ± SD percentage microhardness recovery in remineralized enamel (%REMH) was found in group 4 (56.90 ± 5.49). The %REMH of groups 2 (30.74 ± 3.51) and 5 (29.23 ± 5.65) were statistically the same (p = 0.943). FESEM images confirmed partial coverage of the porous demineralized enamel with a newly formed mineralized layer. Based on EDS findings, the Ca/P ratio values of the treated enamel surfaces with CS-based hydrogels ranged between 1.71 and 1.87, and the highest F content was noticed in group 2 (1.02 ± 0.03). Although, all tested CS-based hydrogels demonstrated the potential to repair demineralized enamel, nHA- and NaF-containing CS-based hydrogel showed the highest remineralization effect. We infer that this new hybrid hydrogel is a potentially useful dental material for tooth biomineralization.

In-vitro and in-vivo comparative studies of treatment effects on enamel demineralization during orthodontic therapy: implications for clinical early-intervention strategy.

OBJECTIVES: This study aimed to investigate if CPP-ACP / infiltrating resin was superior in treating enamel demineralization during orthodontic therapy compared with fluoride varnish, in order to provide early-intervention implications for dental professionals. MATERIALS AND METHODS: In the in-vitro study, premolars were grouped into four: remineralization with fluoride varnish / CPP-ACP, sealing with infiltrating resin, and negative control. Experimental demineralization of enamel surfaces was analyzed using techniques of QLF, SEM, EDS and micro-hardness testing. An in-vivo intervention study was conducted on patients randomly assigned into three groups. At the baseline and every-3-month follow-up, QLF parameters were compared temporally and parallelly to yield potential implications for promotion in clinical practice. RESULTS: The in-vitro study performed on 48 experimental tooth surfaces demonstrated that sealing with infiltrating resin reduced enamel surface porosity and increased surface micro-hardness significantly. In the in-vivo intervention study on 163 tooth surfaces, it was suggested that for those who meet the criteria of -10 < ΔF < -6 and - 1000 < ΔQ < -20 at the baseline, all these treatment methods could achieve acceptable outcomes; with the rising of absolute values of ΔF and ΔQ, sealing with infiltrating resin showed more evident advantages. CONCLUSION: For enamel demineralization during orthodontic therapy, all the treatment methods involved in this study showed acceptable effectiveness but had respective characteristics in treatment effects. QLF parameters could be used as indicators for clinical early-intervention strategy with regards to this clinical issue. CLINICAL RELEVANCE: With QLF parameters, clinical early-intervention strategy for enamel demineralization during orthodontic therapy could be optimized.

New Generation of Orthodontic Elastomeric Ligature to Prevent Enamel Demineralization In Vivo.

This study aimed to synthesize a novel elastomeric ligature with dimethylaminohexadecyl methacrylate (DMAHDM) grafted, providing a new strategy for improving the issue of enamel demineralization during fixed orthodontics. DMAHDM was incorporated into elastomeric ligatures at different mass fractions using ultraviolet photochemical grafting. The antibacterial properties were evaluated and the optimal DMAHDM amount was determined based on cytotoxicity assays. Moreover, tests were conducted to evaluate the in vivo changes in the mechanical properties of the elastomeric ligatures. To assess the actual in vivo effectiveness in preventing enamel demineralization, a rat demineralization model was established, with analyses focusing on changes in surface microstructure, elemental composition, and nanomechanical properties. Elastomeric ligatures with 2% DMAHDM showed excellent biocompatibility and the best antibacterial properties, reducing lactic acid production by 65.3% and biofilm bacteria by 50.0% within 24 h, without significant mechanical property differences from the control group (p > 0.05). Most importantly, they effectively prevented enamel demineralization in vivo, enhancing elastic modulus by 73.2% and hardness by 204.8%. Elastomeric ligatures incorporating DMAHDM have shown great potential for application in preventing enamel demineralization, providing a new strategy to solve this issue during fixed orthodontics.

Remineralizing effect of NSF on artificial enamel caries.

INTRODUCTION: Nanotechnology offers new approaches and endless opportunities for remineralizing tooth decay without being toxic or causing allergies. This study aimed to determine the effect of nanosilver fluoride (NSF) on the remineralization potential of enamel caries-like lesions compared to 5% sodium fluoride varnish in permanent teeth. METHODS: Fifteen teeth (molars and premolars) were gathered, cleaned, and polished using a scaler. After sectioning the teeth mesiodistally and removing the roots, the thirty specimens were subjected to a demineralized solution to induce early enamel lesions and then assigned randomly into two equal groups. The test materials were applied, and then all the specimens were subjected to a pH cycling model for 30 days. DIAGNOdent and surface roughness were investigated, and an evaluation of the enamel Ca and P weight% for Ca/P ratio calculation was done using SEM-EDX to analyze the specimens at the end of the study. The data were analyzed using an independent t-test. RESULTS: The mean values for the DIAGNOdent measurements for NSF and NaF at baseline and after demineralization were not significantly different (p > 0.05). After treatment, NaF varnish showed a significantly higher mean DIAGNOdent measurement (11.8 ± 5.80) than NSF (4.7 ± 1.6). The mean surface roughness of the NaF group (1.64 ± 0.39) was much higher than NSF's mean surface roughness (1.07 ± 0.21). Specimens treated with NSF had statistically significant smoother surfaces (p < 0.001). The NSF group had a higher mean Ca/P ratio (2.9 ± 0.35) than NaF (2.2 ± 0.11). This difference was statistically significant (p = 0.012). CONCLUSION: The study reveals that nano silver fluoride is a more effective treatment than sodium fluoride varnish in enhancing teeth's clinical characteristics, particularly in terms of mineral content and surface roughness, suggesting it could be an improved strategy to prevent dental caries and maintain enamel integrity.

Novel antibacterial orthodontic elastomeric ligature with oral biofilm-regulatory ability to prevent enamel demineralization.

OBJECTIVES: To synthesize a novel antibacterial orthodontic elastomeric ligature incorporating dimethylaminohexadecyl methacrylate (DMAHDM) for the first time to prevent enamel demineralization during orthodontic therapy. METHODS: Various mass fractions of DMAHDM (ranging from 0 % to 20 %) were grafted onto commercial elastomeric ligatures using an ultraviolet photochemical grafting method and were characterized. The optimal DMAHDM concentration was determined based on biocompatibility and mechanical properties, and the antibacterial efficacy was evaluated in a whole-plaque biofilm model. TaqMan real-time polymerase chain reaction and fluorescence in situ hybridization were used to assess the microbial regulatory ability of the multispecies biofilms. Furthermore, an in vitro tooth demineralization model was established to explore its preventive effects on enamel demineralization. Statistical analysis involved a one-way analysis of variance and LSD post hoc tests at a significance level of 0.05. RESULTS: The elastomeric ligature containing 2 % mass fraction of DMAHDM exhibited excellent mechanical properties, favorable biocompatibility, and the most effective antibacterial ability against microorganisms, which decreased by almost two logarithms (P < 0.05). It significantly reduced the proportion of Streptococcus mutans in the multispecies plaque biofilm by 25 % at 72 h, leading to an enhanced biofilm microenvironment. Moreover, the novel elastomeric ligature demonstrated an obvious preventive effect on enamel demineralization, with an elastic modulus 30 % higher and hardness 62 % higher than those of the control group within 3 months (P < 0.05). SIGNIFICANCE: The integration of DMAHDM with an elastomeric ligature holds significant promise for regulating biofilms and preventing enamel demineralization in orthodontic applications.

The combination of arginine and fluoride-containing bioactive glass acted synergistically in inhibiting enamel demineralization in permanent teeth.

OBJECTIVES: To evaluate the in-vitro efficacy of inhibiting enamel demineralization using arginine in combination with fluoride-containing bioactive glass (FBG). METHODS: In this study, the healthy enamel blocks were first demineralized in acetic acid for 24 h, then soaked in anti-demineralization treatment solutions containing either arginine or FBG or both for 96 h.The specimens treated in acetic acid were applied as the control group. The pH, calcium and phosphorus ion concentrations of the solutions were measured before and after treatment. Changes in enamel mineral weight, microhardness, and composition were also analyzed. RESULTS: The present of arginine facilitated fluorine release from treatment solutions with the presence of FBG. Both arginine and FBG significantly increased the pH of treatment solutions and prevented the further mineral weight loss compared to the control group. All anti-demineralization treatment groups showed significant increases in microhardness, but there was no statistical difference among the treatment groups. The SEM analysis showed enamel restoration in the arginine and FBG groups upon treatment, while the combined groups showing a superior anti-demineralization efficacy. 19F NMR showed the formation of fluorapatite in samples treated with solutions containing FBG. CONCLUSIONS: Both arginine and FBG could inhibit enamel demineralization to some extent, and their combination demonstrated an enhanced anti-demineralization efficacy. The low-concentration combination group exhibited anti-demineralization effects comparable to those of high-concentration ones. CLINICAL SIGNIFICANCE: This study introduces a new approach for caries prevention by combining the application of arginine and FBG. The release of fluorine promoted by the presented arginine along with calcium and phosphorus ions from FBG facilitated FAP formation. Additionally, the increment of pH resulting from arginine and FBG degradation further prevents enamel demineralization.

Comparing Shear Bond Strength Between Pink Opaquer and Other Tooth-Colored Restorative Materials on Demineralized Dentin Treated with Silver Diammine Fluoride.

Purpose: The purposes of this study were to evaluate the effect of silver diammine fluoride (SDF) on the shear bond strength (SBS) of pink opaquer (PO) compared to resin-modified glass ionomer (RMGI) and conventional composite (COMP) on demineralized dentin, and also to investigate the mode of failure (MOF). Methods: Sixty extracted third molars were prepared, demineralized for 14 days, and divided into four groups: (1) COMP; (2) SDF+PO; (3) SDF+RMGI; and (4) SDF+COMP (restoration size: two by two mm). SBS, MOF, modified adhesive remnant index (MARI), and remnant adhesive volume (RAV) were evaluated using an Instron® machine, light microscopy, 3D digital scanner ( 3Shape©), and GeoMagic Wrap© software. Results: There was no significant difference in SBS (MPa) among the COMP mean??standard deviation (2.5±1.59), SDF+COMP (2.28±1.05), SDF+PO (3.31±2.63), and SDF+RMGI groups (3.74±2.34). There was no significant difference in MOF and MARI among the four groups (P>0.05). There was no significant difference in RAV (mm3) among the COMP (0.5±0.33), SDF+COMP (0.39±0.44), SDF+PO (0.42±0.38), and SDF+RMGI groups (0.42±0.38; P>0.05). A significant correlation existed between MOF and RAV (R equals 0.721; P<0.001). MOF, MARI, and RAV did not show any correlations with SBS (P>0.05). Conclusions: Silver diammine fluoride does not affect shear bond strength between carious dentinal surface and tooth color restorative materials. The amount of material left on the interface is not related to the amount of shear force needed to break the restoration.

In vitro effect of TiF4/NaF solution on the development of radiation-induced dentin caries.

OBJECTIVE: To evaluate the protective effect of an experimental solution containing TiF4/NaF on the development of radiation-induced dentin caries lesions. METHODOLOGY: bovine root samples were irradiated (70Gy) and distributed as following (n=12/group): Commercial Saliva (BioXtra), NaF (500 ppm F-), TiF4 (500 ppm F), TiF4/NaF (TiF4: 300 ppm F-, NaF: 190 ppm F-), and Phosphate buffer solution (PBS, negative control). Biofilm was produced using biofilm from irradiated patients and McBain saliva (0.2% of sucrose, at 37oC and 5% CO2) for five days. The treatments were applied 1x/day. Colony-forming units (CFU) were counted and demineralization was quantified by transversal microradiography. The ANOVA/Tukey test was applied for all parameters. RESULTS: All treatments reduced CFU for total microorganisms. TiF4 reduced Lactobacillus sp. (7.04±0.26 log10 CFU/mL) and mutans streptococci (7.18±0.28) CFU the most, when compared to PBS (7.58±0.21 and 7.75±0.17) and followed by NaF (7.12±0.31 and 7.34±0.22) and TiF4/NaF (7.16±0.35 and 7.29± 0.29). TiF4 and Commercial saliva showed the lowest integrated mineral loss (ΔZ-vol%.mm) (1977±150 and 2062±243, respectively) when compared to PBS (4540±335), followed by NaF (2403±235) and TiF4/NaF (2340±200). Commercial saliva was the only to significantly reduce mineral loss (LD-µm) (111±25) compared to PBS (153±24).Mean mineral loss (R-vol%) decreased by 35.2% for TiF4 (18.2±3.3) when compared to PBS (28.1±2.9) Conclusion: TiF4/NaF has a comparable anti-cariogenic effect to TiF4 and Commercial saliva under the model in this study.

Protecting primary teeth from dental erosion through bioactive glass.

OBJECTIVES: The present study aimed to evaluate the effectiveness of bioactive glass (BAG) in preventing dental erosion in primary teeth. METHODS: Enamel and dentin specimens (2 × 2 × 2 mm) were obtained from extracted primary teeth, which were randomly divided into the following groups based on the pretreatments (n = 12): DW (deionized water), NaF (2 % sodium fluoride), 2BAG (2 % BAG), 4BAG (4 % BAG), 6BAG (6 % BAG), and 8BAG (8 % BAG). The specimens were immersed in the respective solutions for 2 min and subjected to in vitro erosive challenges (4 × 5 min/d) for 5 d. The erosive enamel loss (EEL), erosive dentin loss (EDL), and the thickness of the demineralized organic matrix (DOM) were measured using a contact profilometer. The surface microhardness (SMH) was measured, and the percentage of SMH loss (%SMHL) was calculated. The surface morphology and mineral composition were evaluated by scanning electron microscopy (SEM) and energy-dispersive X-ray spectroscopy (EDS), respectively. RESULTS: After the erosive challenges, the EEL, EDL, and%SMHL of the 2BAG, 4BAG, 6BAG, and 8BAG groups significantly reduced, with the greatest reduction was observed in the 6BAG (EEL: 6.5 ± 0.2 µm;%SMHL in enamel: 12.8 ± 2.6; EDL: 7.9 ± 0.3 µm; %SMHL in dentin: 22.1 ± 2.7) and 8BAG groups (EEL: 6.4 ± 0.4 µm;%SMHL in enamel: 11.0 ± 1.9; EDL: 7.8 ± 0.5 µm; %SMHL in dentin: 22.0 ± 2.5) (P < 0.05). With increasing BAG concentrations, the number of surface deposits containing Ca, P, and Si increased. CONCLUSIONS: 6BAG was the most effective for preventing dental erosion in primary teeth and showed a particularly strong potential for dentin erosion prevention. CLINICAL SIGNIFICANCE: Bioactive glass, especially at a 6 % concentration, has proven effective in reducing erosive tooth wear and surface microhardness loss while also protecting demineralized organic matrix in primary dentin.

Efficacy of peptide-based enamel coatings in the prevention of demineralization using fixed orthodontic brackets in a rat model.

INTRODUCTION: White spot lesions (WSLs) represent a prominent pathology encountered during orthodontic treatment, originating from enamel demineralization induced by the accumulation of bacterial biofilms. The previously developed bioinspired enamel coating form of self-assembling antimicrobial peptide D-GL13K exhibited antimicrobial activity and enhanced acid impermeability, offering a potential solution to prevent demineralization. The primary aim of this investigation is to assess the in vivo anti-demineralization properties and biocompatibility of the D-GL13K coating. METHODS: A rat model was developed to assess the antimicrobial enamel coating during fixed orthodontic treatment. The anti-demineralization efficacy attributed to the D-GL13K coating was evaluated by employing optical coherence tomography, Vickers microhardness testing, and scanning electron microscopy. The biocompatibility of the D-GL13K coating was investigated through histologic observations of vital organs and tissues using hematoxylin and eosin. RESULTS: The D-GL13K coating demonstrated significant anti-demineralization effects, evidenced by reduced demineralization depth analyzed through optical coherence tomography and enhanced Vickers hardness than in the noncoated control group, showcasing the coating's potential to protect teeth from WSLs. Scanning electron microscopy analysis further elucidated the diminished enamel damage observed in the group treated with D-GL13K. Importantly, histologic examination of vital organs and tissues using hematoxylin and eosin staining revealed no overt disparities between the D-GL13K coated group and the noncoated control group. CONCLUSIONS: The D-GL13K enamel coating demonstrated promising anti-demineralization and biocompatibility properties in a rat model, thereby suggesting its potential for averting WSLs after orthodontic interventions. Further research in human clinical settings is needed to evaluate the coating's long-term efficacy.

Efficacy of the Probiotic L. brevis in Counteracting the Demineralizing Process of the Tooth Enamel Surface: Results from an In Vitro Study.

BACKGROUND: Enamel plays an essential role in protecting the underlying layers of the human tooth; therefore, preserving it is vital. This experimental study aimed to evaluate the potential ability of L. brevis to counteract the action of a demineralizing agent on dental enamel morphology and mineral composition in vitro. METHODS: The sample consisted of 12 healthy human posterior teeth. The coronal portion of each tooth was subdivided into two equal parts longitudinally. The specimens were randomly divided into four groups: artificial saliva, L. brevis suspension, demineralizing agent (DA), and DA plus L. brevis. Scanning electron microscopy (SEM) and energy-dispersive X-ray spectroscopy (EDS) were used to evaluate the surface micromorphology and the mineral content, respectively. The statistical analysis was conducted using a one-way ANOVA, followed by Tukey's post hoc test. RESULTS: SEM analysis did not highlight significant changes in the enamel microstructure of L. brevis-treated specimens compared to the control. DA-induced damage to the enamel structure was drastically reduced when the specimens were contextually exposed to the probiotic. The treatment with DA substantially reduced the weight % of crucial enamel minerals, i.e., Ca and P. Notably, the probiotic was able to reverse the demineralization process, bringing Ca and P weight % back to basal levels, including the Ca/P ratio. CONCLUSIONS: The findings indicate that L. brevis is able to efficiently protect the dental enamel surface from the damage caused by DA and increase the enamel resistance to demineralization. Overall, L. brevis confirms its efficacy in preventing or counteracting the action of carious lesions through a novel mechanism that protects the tooth surface under a chemical challenge that mimics the caries process.

Inhibition of incipient caries lesion progression by different fluoridated varnishes.

The aim of this in vitro study was to evaluate the potential of different fluoridated varnishes to inhibit the progression of incipient caries lesions after cariogenic challenge. Seventy-five enamel specimens of bovine teeth were prepared and selected based on the initial surface microhardness (SMH). The specimens were first subjected to artificial demineralization (in buffer solution) after which SMH was re-analyzed (SM1). They were then randomly assigned to five experimental groups: 1- CONTROL (pH cycling), 2 - MI VAR (MI Varnish with RECALDENTTM - CPP-ACP), 3 - PROFL (Profluorid®), 4 - CLIN (ClinproTM White Varnish with TCP), and 5 - DUR (Duraphat®) (n=15). The varnishes were applied in a thin layer and the specimens were then subjected to pH cycling for eight days. The SMH and cross-sectional microhardness (CSMH) were then analyzed (SM2). The fluoride and calcium ion concentrations in the solution were analyzed by the indirect method and atomic absorption spectrophotometry, respectively. Data were statistically analyzed by Student's t-test, ANOVA/Tukey-Kramer, or Kruskall-Wallis/Dunn tests for individual comparisons (p˂0.05). All varnishes led to significantly higher surface and subsurface remineralization compared with the control group but did not differ from each other. The varnishes with the highest fluoride release were: PROFL and CLIN, followed by MI VAR and DUR. The varnishes with significantly higher release of calcium were: DUR, CLIN, and PROFL. In conclusion, all commercial fluoridated varnishes tested have good potential to inhibit the progression of demineralization, regardless of the ion release mechanisms.

Enhanced anti-biofilm and anti-caries potential of arginine combined with calcium glycerophosphate and fluoride.

OBJECTIVE: The aim of this work was to evaluate the antibiofilm and anticaries properties of the association of arginine (Arg) with calcium glycerophosphate (CaGP) and fluoride (F). METHODS: An active attachment, polymicrobial biofilm model obtained from saliva and bovine teeth discs were used. After the initial biofilm growth period, the enamel discs were transferred to culture medium. The treatment solutions were added to the culture media to achieve the desired final concentration. The following groups were used: negative control (Control); F (110 ppm F); CaGP (0.05 %); Arg (0.8 %) and their associations (F + CaGP; Arg + F; Arg + CaGP; Arg +F + CaGP). The following analyses were carried out: bacterial viability (total bacteria, aciduric bacteria and mutans streptococci), pH assessment of the spent culture medium, dry weight quantification, evaluation of surface hardness loss (%SH) and subsurface mineral content. Normality and homoscedasticity were tested (Shapiro-Wilk and Levene's test) and the following tests were applied: two-way ANOVA (acidogenicity), Kruskall-Wallis (microbial viability) and one way ANOVA (dry weight, %SH, mineral content). RESULTS: The association Arg + F + CaGP resulted in the lowest surface hardness loss in tooth enamel (-10.9 ± 2.3 %; p < 0.05). Arg +F + CaGP exhibited highest values of subsurface mineral content (10.1 ± 2.9 gHAP/cm3) in comparison to Control and F (p < 0.05). In comparison to Control and F, Arg +F + CaGP promoted the highest reduction in aciduric bacteria and mutans streptococci (5.7 ± 0.4; 4.4 ± 0.5 logCFU/mL, p < 0.05). CONCLUSIONS: The Arg-F-Ca association demonstrated to be the most effective combination in protecting the loss of surface hardness and subsurface mineral content, in addition to controlling important virulence factors of the cariogenic biofilm. CLINICAL SIGNIFICANCE: Our findings provide evidence that the Arg-F-Ca association showed an additive effect, particularly concerning protection against enamel demineralization. The combination of these compounds may be a strategy for patients at high risk of caries.

Enamel Matrix Derivative, 58S5 Bioactive Glass, and Fluoride Varnish for Enamel Remineralization: A Multi-analysis Approach.

PURPOSE: This study aimed to evaluate the enamel remineralization efficacy of enamel matrix derivative (EMD), experimental bioactive glass (BAG), and fluoride varnish in vitro. METHODS AND MATERIALS: Artificial initial caries lesions were developed on fifty human enamel specimens using demineralization solution (pH 4.5, 37°C, 96 hours). Specimens were randomly assigned to five groups (n=10): I-5% NaF varnish (Enamelast), II-experimental 58S5 BAG+37% phosphoric acid (PA), III-EMD (Emdogain) + Ethylenediaminetetraacetic Acid (EDTA), IV-EMD+37% PA, V-Control (untreated). All remineralization agents were applied with pH cycling for seven days. The specimens were scanned by spectral domain optical coherence tomography (SD-OCT) at baseline, at demineralization, and after pH cycling. Lesion depths were measured using image analysis software (ImageJ). Lesions were evaluated using surface microhardness (SMH) and two fluorescence methods (FluoreCam and DIAGNOdent Pen [DDPen]). The data were statistically analyzed by Kruskal Wallis, Friedman, and Wilcoxon tests (α=0.05). RESULTS: According to SD-OCT results, fluoride varnish was found to be the most effective agent in reducing lesion depth (p=0.005). All agents increased the SMH values after pH cycling. No significant difference was found among fluoride varnish, BAG, and EMD+PA groups. These SMH values were significantly higher than EMD+EDTA and control groups (p<0.001). All groups showed lower DDPen scores compared with the control group (p<0.001), however, no significant difference was found among the remineralization agents. In FluoreCam assessment, size and intensity values of all treated groups showed improvement. However, there was no significant difference between the treatment groups in terms of FluoreCam size measurements (p=0.186). CONCLUSION: 58S5 BAG and EMD+PA have remineralization capacity as effective as fluoride varnish. EMD+PA showed better SMH and lesion intensity results than EMD+EDTA.

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.

Acquired enamel pellicle and biofilm engineering with a combination of acid-resistant proteins (CaneCPI-5, StN15, and Hemoglobin) for enhanced protection against dental caries - in vivo and in vitro investigations.

OBJECTIVE: This study was designed in two-legs. In the in vivo, we explored the potential of a rinse solution containing a combination (Comb) of 0.1 mg/mL CaneCPI-5 (sugarcane-derive cystatin), 1.88 × 10- 5M StN15 (statherin-derived peptide) and 1.0 mg/mL hemoglobin (Hb) to change the protein profile of the acquired enamel pellicle(AEP) and the microbiome of the enamel biofilm. The in vitro, was designed to reveal the effects of Comb on the viability and bacterial composition of the microcosm biofilm, as well as on enamel demineralization. MATERIALS AND METHODS: In vivo study, 10 participants rinsed (10mL,1 min) with either deionized water (H2O-control) or Comb. AEP and biofilm were collected after 2 and 3 h, respectively, after rinsing. AEP samples underwent proteomics analysis, while biofilm microbiome was assessed via 16 S-rRNA Next Generation Sequencing(NGS). In vitro study, a microcosm biofilm protocol was employed. Ninety-six enamel specimens were treated with: 1)Phosphate-Buffered Solution-PBS(negative-control), 2)0.12%Chlorhexidine, 3)500ppmNaF and 4)Comb. Resazurin, colony-forming-units(CFU) and Transversal Microradiography(TMR) were performed. RESULTS: The proteomic results revealed higher quantity of proteins in the Comb compared to control associated with immune system response and oral microbial adhesion. Microbiome showed a significant increase in bacteria linked to a healthy microbiota, in the Comb group. In the in vitro study, Comb group was only efficient in reducing mineral-loss and lesion-depth compared to the PBS. CONCLUSIONS: The AEP modification altered the subsequent layers, affecting the initial process of bacterial adhesion of pathogenic and commensal bacteria, as well as enamel demineralization. CLINICAL RELEVANCE: Comb group shows promise in shaping oral health by potentially introducing innovative approaches to prevent enamel demineralization and deter tooth decay.

Arginine and sodium fluoride affect the microbial composition and reduce biofilm metabolism and enamel mineral loss in an oral microcosm model.

OBJECTIVE: To assess the effects of arginine, with or without sodium fluoride (NaF; 1,450 ppm), on saliva-derived microcosm biofilms and enamel demineralization. METHODS: Saliva-derived biofilms were grown on bovine enamel blocks in 0.2 % sucrose-containing modified McBain medium, according to six experimental groups: control (McBain 0.2 %); 2.5 % arginine; 8 % arginine; NaF; 2.5 % arginine with NaF; and 8 % arginine with NaF. After 5 days of growth, biofilm viability was assessed by colony-forming units counting, laser scanning confocal microscopy was used to determine biofilm vitality and extracellular polysaccharide (EPS) production, while biofilm metabolism was evaluated using the resazurin assay and lactic acid quantification. Demineralization was evaluated by measuring pH in the culture medium and calcium release. Data were analyzed by Kruskal-Wallis' and Dunn's tests (p < 0.05). RESULTS: 8 % arginine with NaF showed the strongest reduction in total streptococci and total microorganism counts, with no significant difference compared to arginine without NaF. Neither 2.5 % arginine alone nor NaF alone significantly reduced microbial counts compared to the control, although in combination, a reduction in all microbial groups was observed. Similar trends were found for biofilm vitality and EPS, and calcium released to the growth medium. CONCLUSIONS: 8 % Arginine, with or without NaF, exhibited the strongest antimicrobial activity and reduced enamel calcium loss. Also, NaF enhanced the effects of 2.5 % arginine, yielding similar results to 8 % arginine for most parameters analyzed. CLINICAL SIGNIFICANCE: The results provided further evidence on how arginine, with or without NaF, affects oral microcosm biofilms and enamel mineral loss.

Clinical and ex-vivo effect of LASERs on prevention of early-enamel caries: systematic review & meta-analyses.

To investigate the in vivo and in situ effect of different types of lasers in prevention of enamel demineralization in high caries risk cases (around orthodontic brackets, around restoration and in caries susceptible pits and fissures). PubMed was searched using the following keyword sequence; (Laser therapy OR laser irradiation OR laser application) AND (enamel caries prevention OR enamel demineralization OR enamel remineralization OR early enamel caries OR early-enamel caries OR enamel resistance OR enamel decalcification OR white spot lesions WSLs OR incipient lesion OR enamel decay OR enamel Dissolution OR enamel microhardness) AND (clinical trial OR Randomized clinical trial OR In situ study). The latest literature search was ended by "30 January 2023". PubMed was used as a primary data base for study selection. Scopus, EBSCO, and Google scholar are checked in our study after results of systematic search on PubMed. Only duplicates were found. Two meta-analyses were carried out. The first, clinical meta-analysis on incidence of white spot lesions (WSLs) following CO2 laser irradiation of enamel. The second meta-analysis on ex-vivo/in situ effect of CO2 laser on microhardness of enamel. In each meta-analysis three studies were included. Risk of bias was assessed. The search identified eight studies (four ex-vivo and four clinical trials). Regarding the clinical meta-analysis, the overall standardized mean difference was 0.21 [ 95% confidence interval (CI): 0.15-0.30, p < 0.00001]. This indicates that the incidence of new WSLs in patients who received low power CO2 laser treatment was highly significantly lower than placebo groups. The heterogeneity was considerable (I2 = 71%). In the second meta-analysis, the overall standardized mean difference was 49.55 [ 95% confidence interval (CI): 37.74, 61.37, p < 0.00001]. This indicates that microhardness of enamel receiving low power (0.4-5 W) CO2 laser irradiation is highly significantly lower than control untreated enamel. The heterogeneity was substantial (I2 = 48%). Within the limitations of this study, Low level laser therapy concept with CO2 laser seems to be effective in preventing enamel caries.Prospero registration number: CRD42023437379.

Comparison of the effects of fluoride varnish containing silver nanoparticles and conventional fluoride varnish on the surface microhardness of tooth enamel.

BACKGROUND: Nano-silver fluoride (NSF) has been introduced to improve enamel lesions. The effective use of varnishes is important in the prevention of dental caries. OBJECTIVES: The study aimed to compare the effect of conventional sodium fluoride varnish with the same varnish containing 1% and 2% silver nanoparticles (AgNP) on the surface microhardness of enamel. MATERIAL AND METHODS: The baseline surface microhardness of 40 premolar teeth was measured using a Vickers microhardness tester. After immersing the samples in a demineralizing agent for 24 h, the microhardness was measured again. In group B, a layer of conventional fluoride varnish was applied to the tooth surfaces using a microbrush with soft bristles, following the manufacturer's instructions. Groups C and D were treated with 1% and 2% NSF varnishes, respectively, while group A received no varnish. Surface microhardness tests were conducted on all specimens, including those previously tested. RESULTS: The microhardness of the enamel surface increased significantly in all 3 test groups compared to the microhardness after demineralization (p < 0.05). CONCLUSIONS: Conventional fluoride varnish and fluoride varnishes containing 1% and 2% AgNP are equally effective in remineralizing initial caries.

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.