OBJECTIVE: In this in vivo proof-of-concept study, acquired pellicle engineering was implemented to promote alterations in the protein composition of the acquired enamel pellicle (AEP) and the bacterial composition of the dental biofilm after treatment with Sugarcane cystatin (CaneCPI-5).
METHODS: After prophylaxis, 10 volunteers rinsed (10 mL, 1 min) with the following solutions: 1) deionized water (H2O- negative control or 2) 0.1 mg/mL CaneCPI-5. The AEP and biofilm were formed along 2 or 3 h, respectively. The AEP was collected with electrode filter papers soaked in 3 % citric acid. After protein extraction, samples were analyzed by quantitative shotgun label-free proteomics. The biofilm microbiome was collected with a dental curette. The DNA was extracted, amplified, and analyzed by 16S-rRNA Next Generation Sequencing (NGS).
RESULTS: Treatment with CaneCPI-5 increased several proteins with antimicrobial, acid-resistance, affinity for hydroxyapatite, structural and calcium binding properties, such as Cysteine-rich-3 (6-fold-p = 0.03), Cystatin-B (5.5-fold-p < 0.01), Neutrophil-defensin 1 (4.7-fold-p < 0.01), Mucin (3.9-fold-p < 0.01), Immunoglobulin-heavy-constant (3.8-fold-p < 0.01) and Lactotransferrin (2.8-fold-p < 0.01). Microbiome revealed that several commensal bacteria had their abundance increased after rinsing with CaneCPI-5, such as Corynebacterium and Neisseria, while Streptococcus and Prevotella nigrescens were decreased. The results indicate the efficiency of CaneCPI-5 in promoting beneficial changes in the AEP and biofilm, making this phytocystatin a potential target for incorporation into dental products.
CONCLUSIONS: Cane demonstrated the capability to alter the protein composition of the acquired enamel pellicle (AEP) and the initial colonizers of the biofilm, enhancing the presence of proteins and bacteria crucial for dental protection.

OBJECTIVE: To develop a protocol for forming subsurface caries lesions on bovine enamel by dual-species biofilms of Streptococcus mutans and Candida albicans in vitro.
METHODS: Biofilms were grown on bovine enamel specimens in artificial saliva (AS) for seven days. After 24 h of formation, the AS was supplemented or not with fluoride (F) using sodium fluoride (0.005 or 0.008 ppm F), and the biofilms were exposed or not to a 20 % sucrose solution (reproducing a cariogenic challenge) once/day. On the seventh day, the biofilms were harvested and had their extracellular polysaccharides (EPS) and inorganic components analyzed. The specimens were subjected to computed X-ray microtomography analysis to determine their mineral concentration. Data were compared using two-way analyses of variance, followed by Fisher\'s LSD or Student-Newman-Keuls tests (p < 0.05).
RESULTS: Biofilms exposed to the cariogenic challenge had significantly higher EPS concentrations than those not exposed, regardless of the presence of F. For biofilms grown with 0.008 ppm F, those exposed to the cariogenic challenge had lower F levels than those not exposed. For biofilms exposed to the cariogenic challenge, those grown with 0.008 ppm F had lower lesion depths and integrated mineral loss, and higher outer layers than those grown without F.
CONCLUSIONS: The dual biofilm model assessed was able to create subsurface caries lesions in bovine enamel in vitro, which was influenced by the presence of F in the culture medium and exposure to sucrose.

OBJECTIVE: Sugary drinks such as Coca-Cola may expedite dental caries. For this reason, sugar-free drinks like Coca-Cola Zero Sugar (CZ) may be considered advantageous. This research aims to evaluate in vitro the CZ effect in the presence of Streptococcus mutans (S. mutans) biofilm on enamel demineralization.
METHODS: Ninety-six human enamel slabs (4 × 4 mm) were used. S. mutans UA-159 72-hour biofilm was created over enamel surfaces. The specimens were soaked in CZ, HCl, or 10% sucrose in PBS solution, 3 times a day for 15 minutes over the course of 4 days. Viable counts (CFU/mL) and biofilm biomass (Crystal Violet staining) were evaluated. pH was measured after each exposure. After 4 days, Demineralization was evaluated clinically and by Vickers microhardness tests. Slabs were photographed using a stereomicroscope before and after exposure to caries-promoting conditions.
RESULTS: Slabs that were soaked in CZ showed an increase in viable counts compared to control and almost similar counts with 10% sucrose in PBS solution exposures (1010and109CFUmL, respectivly). Biofilm biomass tests showed a 25% higher bacterial growth in the CZ group. CZ pH measures were the lowest and the only group to show a decrease in pH over time (pH ∼3). Enamel slabs that were evaluated clinically in the stereomicroscope postexposures had a chalky and matt appearance as opposed to their shiny appearance in the baseline evaluation.
CONCLUSIONS: CZ creates a favourable environment for the growth of S. mutans. It may be suggested that even though CZ is sugar free it has a cariogenic effect on enamel.
CONCLUSIONS: Clinicians need to educate patients that sugar-free carbonated drinks may be just as harmful as regular carbonated drinks, and hence avoided. This research emphasizes the harmful effect sugar-free carbonated drinks on teeth and sheds new light on their cariogenic potential.

UNASSIGNED: The current study\'s aim was to evaluate the potential of various fluoride varnishes to remineralize artificial enamel lesions on primary teeth.
UNASSIGNED: For the study, 90 deciduous molar teeth that were implicated for extraction were taken. A window region with a size of 3 × 3 mm was located in the middle of the tooth\'s coronal surface. The remaining portion was given a coat of nail polish and given time to dry. A demineralizing solution was used to create the artificial enamel lesion. The samples were randomized into three experimental groups at random, with 30 samples in each group. Group 1: Application of Clinpro White Varnish, Group 2: Application of Duraphat Varnish, and Group 3: Application of MI Varnish. After 14 days, samples from each group were examined under scanning electron microscopy at a magnification of 1000× to determine whether remineralization had occurred. Microhardness was also determined using a microhardness tester.
UNASSIGNED: Before the intervention, the mean demineralized enamel lesion in the Clinpro White Varnish group was 134.44 ± 0.04, in the Duraphat Varnish group was 133.76 ± 0.12, and in the MI Varnish group was 133.89 ± 0.08. After intervention, the maximum remineralized area was found in the MI Varnish group (82.74 ± 0.18) followed by the Clinpro White Varnish group (101.43 ± 0.16) and the Duraphat Varnish group (104.11 ± 0.10). After the intervention, there was a statistically significant difference found between the groups. After the intervention, the maximum microhardness mean value improved in the MI Varnish group (257.03 ± 1.09 to 261.18 ± 1.20) followed by Clinpro White Varnish (258.78 ± 1.64 to 260.10 ± 1.22) and Duraphat Varnish group (255.24 ± 1.51 to 258.02 ± 0.89).
UNASSIGNED: According to the findings of the current investigation, all of the varnishes utilized in this in vitro experiment are capable of correcting early enamel defects on primary teeth. However, MI Varnish group was very effective compared to the Clinpro White Varnish group and the Duraphat Varnish group.

OBJECTIVE: Protocols that determine the lesion depth of specific demineralization solutions are lacking. This in vitro study aimed to evaluate various lesion depths of artificial white spot lesions (WSLs) at different exposure times.  Materials and methods: Artificial WSLs were created by placing 30 extracted human premolar teeth into 0.05M acetate buffer solution with a controlled environment of pH 4.4 at 37ºC, which were then exposed in the solution for various durations of 4, 5, 6, 8, 10, and 12 days. The specimens were visually examined using the Ekstrand-Ricketts-Kidd (ERK) system to confirm the WSL, followed by buccolingual sectioning, and evaluated under a scanning electron microscope (SEM) to measure the lesion depth.
RESULTS: The SEM showed that the mean lesion depths of representative specimens were 101.33 µm (day 4), 124 µm (day 5), 159 µm (day 6), 187 µm (day 8), 202 µm (day 10), and 212 µm (day 12). The artificial WSL was visually demonstrated in grades 1 and 2 of the ERK system.
CONCLUSIONS: The depths of the lesions increased as the duration increased from day 4 to day 12, indicating that the lesion depths increased with the more prolonged exposure to the acetate buffer solution.

Aim The aim of the study was to compare the effectiveness of chitosan nanoparticle pretreatment with four different remineralizing agents in artificial white spot lesions. Methods  A total of 100 human maxillary first premolars were selected and divided into five groups of 20 samples in each group. Artificial white spot lesions were created by immersing the samples in the demineralizing solution for 96 hours. Chitosan pretreatment was done for all samples followed by subjecting Group I samples to artificial saliva (control), Group II samples to 3M Clinpro, Group III samples to GC Tooth Mousse, Group IV samples to SHY-NM, and Group V samples with Aclaim using a cotton applicator tip. Each group was divided into two subgroups of 10 samples, which were subjected to hardness testing and mineral content analysis. Surface microhardness and the calcium phosphorous ratio were recorded using a Vickers microhardness tester and energy-dispersive X-ray (EDAX) analysis at three levels i.e., baseline, after demineralization, and after remineralization and tabulated. Statistical analysis was conducted by analyzing data using ANOVA and post hoc followed by Dunnett\'s t-test using IBM SPSS Statistics for Windows, Version 16 (Released 2007; IBM Corp., Armonk, New York, United States). Results Vickers surface hardness testing and EDAX analysis showed statistically significant values for all the groups. Among them, maximum remineralization potential was seen in samples treated with Chitosan and 3M Clinpro combination, and minimum remineralization potential was seen in Chitosan and artificial saliva combination. Conclusion The addition of chitosan nanoparticles with various remineralizing agents showed a significant synergistic effect on remineralization activity. Also, chitosan and Clinpro combination showed the maximum surface hardness and EDAX analysis values when compared to other groups.

OBJECTIVE: The gaps at the margins of restorative composite resin can increase as the carious process occurs underneath the materials, causing further demineralization along the tooth cavity wall. The aim of this study was to evaluate the effects of restorative resin composite containing hydrated calcium silicate (hCS) filler on enamel protection against demineralization by simulating microleakage between the test material and teeth in a cariogenic environment.
METHODS: The experimental resin composites were composed of 70 wt.% filler, which was mixed with a glass filler and hCS in a weight ratio of 70.0% glass (hCS 0), 17.5% hCS + 52.5% glass (hCS 17.5), 35.0% hCS + 35.0% glass (hCS 35.0), and 52.5% hCS + 17.5% glass (hCS 52.5). A light-cured experimental resin composite disk was positioned over a polished bovine enamel disk, separated by a 30-µm gap, and immersed in artificial saliva with pH 4.0 for 15, 30, and 60 days. After the immersion period, the enamel disk was separated from the resin composite disk and evaluated using a microhardness tester, atomic force microscopy, and polarized light microscopy. The opposing sides of the enamel and resin composite disks were observed using scanning electron microscopy/energy dispersive X-ray spectrometry.
RESULTS: The enamel surface showed a significant increase in microhardness, decreased roughness, and remineralization layer as the proportion of hCS increased (P < .05). In the scanning electron microscopy image, the enamel surface with hCS 35.0 and 52.5 after all experimental immersion periods, showed a pattern similar to that of a sound tooth.
CONCLUSIONS: The results demonstrated that increasing the hCS filler level of restorative resin composites significantly decreased enamel demineralization.
CONCLUSIONS: Hydrated calcium silicate laced restorative resin composites may be a promising dental biomaterial for protecting teeth against demineralization and preventing secondary caries around restorations.

UNASSIGNED: To evaluate the remineralization potential of varying concentrations of two plant-based extracts of Cocos nucifera on white spot lesions using SEM and EDAX in vitro testing methods.
UNASSIGNED: The pulp was freshly obtained from coconut and divided into two. Then, coconut milk was obtained by blending, while the next portion was freeze-dried and lyophilized. Third molar teeth were processed into tooth slabs (N = 40) and split equally into five groups by block randomization. After demineralization, one tooth slab was taken from each, and SEM analysis was done. Remineralization was then performed among the various groups that included Group 1, which acted as a control and consisted of the remineralization solution. Groups 2 and 3 comprised 1:1 and 2:1 concentrations of the coconut milk, whereas Groups 4 and 5 consisted of 1:1 and 2:1 concentrations of the lyophilized extract. SEM and EDAX testing were done post-remineralization. Ca and phosphate values were tabulated, and statistical significance was determined for the obtained values using ANOVA.
UNASSIGNED: Among the control and treatment groups, surface remineralization was better observed in 1:1 coconut milk and 2:1 coconut milk than in the 2:1 lyophilized coconut, control, and 1:1 lyophilized coconut. Between the control and treatment groups, Ca and phosphate percentages (P < 0.001) showed statistical differences. The lowest value of 2.3% was noted in the 2:1 lyophilized coconut group.
UNASSIGNED: Coconut extracts exhibit remineralization potential on the artificial carious lesion. Coconut milk exhibited significant improvement in the surface properties than lyophilized coconut.

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.

Enamel demineralization is a very common occurrence around bonded brackets in an orthodontic practice. Fluoride (FLR) applications have been used to prevent decalcification and further progression of white spot lesions. The purpose of this systematic review and meta-analysis was to systematically appraise available literature on the effectiveness of fluoride mouthrinse in the prevention of demineralization around fixed orthodontic appliances. A search was conducted for randomized controlled clinical trials among four electronic databases (MEDLINE, Google Scholar, PubMed, and Cochrane Review) through MeSH terms and keywords. Studies were excluded if random allocation was not conducted, or if they were animal or in vitro studies. About 146 articles were screened and 5 studies were selected for the present review. Only two studies were selected for MA due to variations in the measurement of outcomes among studies. This review concluded that rinsing with FLR in the course of the fixed orthodontic treatment lessens demineralization around the bracket. Using FLR mouthrinse to inhibit the formation of white spot lesions or dental caries in patients with multiple cavities or restoration can be considered in clinical practice.