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.

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

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

Pomegranate extract in polyphosphate-fluoride mouthwash reduces enamel demineralization.

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

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

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

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.

Can acid produced from probiotic bacteria alter the surface roughness, microhardness, and elemental composition of enamel? An in vitro study.

Probiotics are live microorganisms that upon administration in adequate amounts provide various health benefits to the host. Probiotics are "lactic acid-producing bacteria" as they release large amounts of organic acids, particularly lactic acids, in their surrounding environment. Although the acids produced by probiotics are beneficial for gastrointestinal and vaginal health, the acidogenic nature of probiotics has raised concerns among dental professionals, especially concerning their effect on the enamel and dentin. Previous studies have found that probiotics can lower the pH of the saliva and cause essential elements like Calcium and Phosphorus to leach from the enamel. This can alter the surface topography of enamel and increase the risk of enamel defects. Studies have also noted that probiotic bacteria can replace cariogenic bacteria and lower the risk of tooth decay. However, the effect of acid produced by probiotics on the enamel surface remains unclear. Hence, the present study aims to evaluate the effect of probiotics on the surface roughness, microhardness, and elemental composition of enamel compared to 0.1 M Lactic acid (demineralizing agent). Twenty enamel sections were randomly divided into groups and subjected to a pH cycling model using a probiotic suspension and 0.1 M lactic acid. The changes in the surface roughness, microhardness, surface morphology, and elemental composition of the enamel with regard to Carbon, Oxygen, Sodium, Hydrogen, Magnesium, Phosphorus, Fluoride, Chlorine, and Calcium of the enamel were evaluated before and after the emersion in both the groups. The results showed a significant increase in the mean surface roughness in the probiotic group before and after the exposure. The microhardness of the enamel decreased along with altered arrangement of the enamel prisms, increased striations, scratch marks, and pitting after exposure to the probiotic group. A decrease in the atomic/weight% for Calcium, Phosphorous, Fluoride, Aluminium, and Oxygen and an increase in the weight/atomic% for Carbon, Nitrogen, and Sodium were noted compared to the baseline in the probiotic solution. The results in the probiotic group were comparable to the 0.1 M lactic acids. The pH changed from 5.78 to 3.06 at the end of 24 h in the probiotic group. Based on these findings, we conclude that exposure to probiotics can affect microhardness and surface roughness and cause leaching of essential elements like Calcium and Phosphorous from the enamel.

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

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

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

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

The cariogenic effect of starch on oral microcosm grown within the dual constant depth film fermenter.

Evidence on the link between starch intake and caries incidence is conflicting, therefore the cariogenicity of starch compared with sucrose was explored using a dual Constant Depth Film Fermenter (dCDFF) biotic model system. Bovine enamel discs were used as a substrate and the dCDFF was inoculated using human saliva. CDFF units were supplemented with artificial saliva growth media at a constant rate to mimic resting salivary flow rate over 14 days. The CDFF units were exposed to different conditions, 2% sucrose or 2% starch 8 times daily and either no additional fluoride or 1450 ppm F- twice daily. Bovine enamel discs were removed at intervals (days 3, 7, 10 and 14) for bacterial enumeration and enamel analysis using Quantitative Light Induced Fluorescence (QLF) and Transverse Microradiography (TMR). Results showed that in the absence of fluoride there was generally no difference in mineral loss between enamel exposed to either sucrose or starch when analysed using TMR and QLF (P > 0.05). In the presence of fluoride by day 14 there was significantly more mineral loss under starch than sucrose when analysed with TMR (P < 0.05). It was confirmed that starch and sucrose are similarly cariogenic within the dCDFF in the absence of fluoride. With the aid of salivary amylase, the bacteria utilise starch to produce an acidic environment similar to that of bacteria exposed to sucrose only. In the presence of fluoride, starch was more cariogenic which may be due to the bacteria producing a more hydrophobic intercellular matrix lowering the penetration of fluoride through the biofilm. This is significant as it indicates that the focus on sugars being the primary cause of caries may need re-evaluating and an increase in focus on carbohydrates is needed as they may be similarly cariogenic as sugars if not more so.

The Effect of Solutions Containing Extracts of Vochysia tucanorum Mart., Myrcia bellaCambess., Matricaria chamomilla L. and Malva sylvestris L. on Cariogenic Bacterial Species and Enamel Caries Development.

This study evaluated the effect of experimental solutions containing plant extracts on bacterial species and enamel caries prevention. Microcosm biofilm was produced from human saliva mixed with McBain saliva (0.2% sucrose) on bovine enamel for 5 days (3 days under anaerobiosis and 2 days under aerobiosis) at 37°C. From the 2nd day, the following treatments were applied (1 × 60 s/day): Vochysia tucanorum (10 mg/mL); Myrcia bella (5 mg/mL); Matricaria chamomilla (80 mg/mL); Malva sylvestris, fluoride, and xylitol (Malvatricin Plus®); 0.12% chlorhexidine (CHX, PerioGard®); and PBS (negative control). The medium pH was measured. Quantitative polymerase chain reaction was performed for the detection of Streptococcus mutans and Lactobacillus spp. Enamel demineralization was measured by spectral-domain optical coherence tomography. The data were compared by means of the Kruskal-Wallis/Dunn, two-way ANOVA/Bonferroni, and ANOVA/Tukey tests (p < 0.05). The pH decreased after sucrose exposure; only CHX reestablished pH >5.5 by the last day. CHX also eliminated Lactobacillusspp., but the other treatments did not differ significantly from PBS. Malvatricin Plus® and CHX eliminated S. mutans, but the other treatments did not differ from PBS. Similar results were seen concerning the reduction of lesion depth and reflectivity. The experimental natural-extract solutions were ineffective against cariogenic bacteria and in preventing the development of enamel caries.

Effect of green tea-loaded chitosan nanoparticles on leathery dentin microhardness.

The purpose of this study was to assess the effect of a chitosan-based nanoformulation containing green tea on leathery (remaining) dentin subsurface microhardness. Size distribution, polydispersity index (PDI) and zeta potential (mV) of nanoformulations were previously determined by dynamic light scattering (DLS). Human dentin specimens were exposed to Streptococcus mutans for 14 d. Soft dentin were selectively removed by Er:YAG laser (n = 30) or bur (n = 30). Remaining dentin was biomodified with chitosan nanoparticles (Nchi, n = 10) or green tea-loaded chitosan nanoparticles (Gt + Nchi, n = 10) for 1 min. Control group (n = 10) did not receive any treatment. Subsurface microhardness (Knoop) was evaluated in hard (sound) and soft dentin, and then, in leathery dentin and after its biomodification, at depths of 30, 60 and 90 µm from the surface. Nchi reached an average size of ≤ 300 nm, PDI varied between 0.311 and 0.422, and zeta potential around + 30 mV. Gt + Nchi reached an average size of ≤ 350 nm, PDI < 0.45, and zeta potential around + 40 mV. Soft dentin showed significantly reduced microhardness at all depths (p > 0.05). The subsurface microhardness was independent of choice of excavation method (p > 0.05). At 30 µm from the surface, Gt + Nchi increased the leathery dentin microhardness compared to untreated group (p < 0.05). Nchi promoted intermediate values (p > 0.05). Both nanoformulations showed an average size less than 350 nm with nanoparticles of different sizes and stability along the 90-day period evaluated. Subsurface microhardness of bur-treated and laser-irradiated dentin was similar. At 30 µm, the biomodification with Gt + Nchi improved the microhardness of leathery dentin, independently of caries excavation method used.

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.

Combined effect of casein phosphopeptide-amorphous calcium phosphate and sodium trimetaphosphate on the prevention of enamel demineralization and dental caries: an in vitro study.

OBJECTIVE: To evaluate the effect of treatment with fluoridated toothpaste supplemented with a combination of sodium trimetaphosphate (TMP) and casein phosphopeptide-amorphous calcium phosphate (MI Paste Plus®) on the demineralization of dental enamel. METHODS: Bovine enamel blocks selected by initial surface hardness (SHi) were randomly allocated into six groups (n = 12), according to the test toothpastes: (1) without F-TMP-MI Paste Plus® (Placebo); (2) 1100 ppm F (1100F); (3) MI Paste Plus®; (4) 1100F + MI Paste Plus® (1100F-MI Paste Plus), (5) 1100F + 3 % TMP (1100F-TMP); and (6) 1100F-TMP + MI Paste Plus® (1100F-TMP-MI Paste Plus). Blocks were treated two times per day with slurries of toothpaste (1 min), and groups 4 and 6 received an application of MI Paste Plus (3 min). Next, the samples were subjected to five pH cycles (demineralizing/remineralizing solutions) at 37 °C, to produce subsurface enamel lesions.Thereafter, the blocks were maintained for 2 days in fresh remineralizing solution. After pH cycling, the following were obtained: percentage of surface hardness loss (%SH); integrated loss of subsurface hardness (ΔKHN); profile analysis and lesion depth subsurface through polarized light microscopy (PLM); scanning electron microscopy (SEM); and fluoride (F), calcium (Ca), and phosphorus (P) in the enamel. The data were subjected to ANOVA (1-criterion), followed by the Student-Newman-Keuls test (p < 0.001). RESULTS: The 1100F-TMP-MI Paste Plus group showed better results for SHR, ΔKHN, and PLM (p < 0.001). The F concentration was similar among all groups (p > 0.001). The 1100F-TMP-MI Paste Plus group showed the highest concentration of Ca and P in the enamel (p < 0.001). CONCLUSION: The application of 1100F-TMP-MI Paste Plus promoted a higher inhibitory effect against enamel demineralization. CLINICAL SIGNIFICANCE: The combination of treatments with F, TMP, and MI Paste Plus® can be an effective alternative to improve the oral health of individuals, especially those with high activity of dental caries and at high risk for its development.

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

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

Qualitative and quantitative assessment of the potential effect of cigarette smoking on enamel of human smokers' teeth.

OBJECTIVE: This study aimed to examine the potential changes in enamel surface of human smokers' teeth. MATERIALS AND METHODS: Forty extracted permanent, human, noncarious anterior teeth were used in this study. Half of these teeth were obtained from heavy smokers, while the other half of teeth were collected from nonsmokers (control teeth). The teeth were then subjected for scanning electron microscopic examination together with energy dispersive X ray and micro-hardness analysis to evaluate the qualitative and quantitative effect of smoking respectively. RESULTS: SEM of smokers' teeth showed variable degrees of destruction from small areas of demineralization as holes and pits to destruction and deterioration of the organizational pattern of the rod substance. Moreover, areas of defective remineralization were detected. The microhardness, calcium and phosphorus weight % significantly decreased whereas the Ca/P ratio was significantly increased. CONCLUSION: Cigarette smoking adversely affected the ultrastructure and mechanical properties of enamel and even hindered the normal remineralization process thus cigarette smoking cessation should be promoted in the dental office daily practices.

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

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

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

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

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

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

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

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