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Item Anti-Caries Efficacy of Fluoride at Increasing Maturation of a Microcosm Biofilm(2019-08) Ayoub, Hadeel Mohammed; Lippert, Frank; Gregory, Richard L.; Martinez Mier, E. Angeles; Anderson, GregoryDental biofilm is a main contributing factor in the initiation and progression of dental caries. The maturation of dental biofilms is expected to alter the anti-caries efficacy of fluoride compounds. In the first aim, we conducted a series of modeldevelopment experiments to test different variables to standardize a reproducible in-vitro microbial caries model. We evaluated: surface conditioning using saliva; sucrose concentrations and caries lesion severity; growth media conditions and mineral saturation; dental substrate types; pH cycling protocol characteristics. In the second aim, we used the developed model to evaluate the changes in the anti-caries efficacy of three fluoride compounds (Sodium fluoride (NaF); Stannous fluoride (SnF2); Amine fluoride (AmF); and deionized water (DIW- negative control)) at increasing maturation of a microcosm biofilm. We continued the pH cycling protocol for 4 days, 8 days, and 12 days. We tested biofilm cariogenicity and carious lesion severity at each maturation stage. In the third aim, we used the developed model to test the effect of different exposure periods (early vs. late exposure) of the biofilm to three fluoride compounds (NaF, SnF2, AmF, DIW) in comparison to DIW. We also evaluated the recovery of biofilm cariogenicity with each exposure period. We evaluated, for each exposure period and recovery stage, biofilm cariogenicity and carious lesion severity. We analyzed the relationships between different variables (biofilm age, fluoride compound type, exposure period) using ANOVA models. In conclusion: 1. The present model allows testing the effect of biofilm maturation on the anti-caries efficacy of fluoride compounds. 2. Biofilm maturation plays an important role in increasing biofilm tolerance against fluoride treatment; it could also influence the selection of fluoride compounds to achieve optimum cariostatic effect. 3. Exposure period, and type of fluoride compound, both influence the biofilm tolerance to fluoride anti-caries effect; they may also result in a sustainable release of fluoride over time.Item Artificial Caries Lesion Characteristics after Secondary Demineralization with Theobromine-Containing Protocol(MDPI, 2021-01-08) Nassar, Hani M.; Lippert, Frank; Cariology, Operative Dentistry and Dental Public Health, School of DentistryDeveloping artificial caries lesions with varying characteristics is needed to adequately study caries process in vitro. The objective of this study was to investigate artificial caries lesion characteristics after secondary demineralization protocol containing theobromine and fluoride. Sixty bovine enamel slabs (4 × 3 mm) were demineralized using a Carbopol-containing protocol for 6 days. A baseline area (2 × 3 mm) was protected with acid-resistant nail varnish, after which specimens were exposed for 24 h to a secondary demineralization protocol containing acetic acid plus one of four fluoride/theobromine combinations (n = 15): theobromine (50 or 200 ppm) and fluoride (0 or 1 ppm). Specimens were sectioned and analyzed using transverse microradiography for changes in mineral content, lesion depth, and surface layer mineralization. Data was analyzed using paired t-test and analysis of variance followed by Bonferroni test at 0.05 significance level. After secondary demineralization, fluoride-containing groups had significantly deeper lesions (p = 0.002 and 0.014) compared to the group with 0 ppm fluoride and 50 ppm theobromine. Mineral content and lesion depth were significantly different compared to baseline for all groups. Theobromine did not show an added effect on mineral uptake. Theobromine-containing groups exhibited particularly deep lesions with a more uniform mineral profile in the presence of fluoride.Item Bioactivity of Dental Restorative Materials: FDI Policy Statement(Elsevier, 2023) Schmalz, Gottfried; Hickel, Reinhard; Price, Richard Bengt; Platt, Jeffrey A.; Biomedical Sciences and Comprehensive Care, School of DentistryThe term bioactivity is being increasingly used in medicine and dentistry. Due to its positive connotation, it is frequently utilised for advertising dental restorative materials. However, there is confusion about what the term means, and concerns have been raised about its potential overuse. Therefore, FDI decided to publish a Policy Statement about the bioactivity of dental restorative materials to clarify the term and provide some caveats for its use in advertising. Background information for this Policy Statement was taken from the current literature, mainly from the PubMed database and the internet. Bioactive restorative materials should have beneficial/desired effects. These effects should be local, intended, and nontoxic and should not interfere with a material's principal purpose, namely dental tissue replacement. Three mechanisms for the bioactivity of such materials have been identified: purely biological, mixed biological/chemical, or strictly chemical. Therefore, when the term bioactivity is used in an advertisement or in a description of a dental restorative material, scientific evidence (in vitro or in situ, and preferably in clinical studies) should be provided describing the mechanism of action, the duration of the effect (especially for materials releasing antibacterial substances), and the lack of significant adverse biological side effects (including the development and spread of antimicrobial resistance). Finally, it should be documented that the prime purpose, for instance, to be used to rebuild the form and function of lost tooth substance or lost teeth, is not impaired, as demonstrated by data from in vitro and clinical studies. The use of the term bioactive dental restorative material in material advertisement/information should be restricted to materials that fulfil all the requirements as described in the FDI Policy Statement.Item Developing an approach to improve beta-phase properties in ferroelectric pvd-hfp thin films(2020-05) Dale, Ashley S.; Cheng, Ruihua; Petrache, Horia; Wassall, StephenImproved fabrication of poly(vinylindenefluoride)-hexafluoropropylene (PVDF-HFP) thin films is of particular interest due to the high electric coercivity found in the beta-phase structure of the thin film. We show that it is possible to obtain high-quality, beta-phase dominant PVDF-HFP thin films using a direct approach to Langmuir-Blodgett deposition without the use of annealing or additives. To improve sample quality, an automated Langmuir-Blodgett thin film deposition system was developed; a custom dipping trough was fabricated, a sample dipping mechanism was designed and constructed, and the system was automated using custom LabVIEW software. Samples were fabricated in the form of ferroelectric capacitors on substrates of glass and silicon, and implement a unique step design with a bottom electrode of copper with an aluminum wetting layer and a top electrode of gold with an aluminum wetting layer. Samples were then characterized using a custom ferroelectric measurement program implemented in LabVIEW with a Keithley picoammeter/voltage supply to confirm electric coercivity properties. Further characterization using scanning electron microscopy and atomic force microscopy confirmed the improvement in thin film fabrication over previous methods.Item Dietary Fluoride Intake during Pregnancy and Neurodevelopment in Toddlers: A Prospective Study in the PROGRESS Cohort(Elsevier, 2021) Cantoral, Alejandra; Téllez-Rojo, Martha M.; Malin, Ashley J.; Schnaas, Lourdes; Osorio-Valencia, Erika; Mercado, Adriana; Martínez-Mier, E. Ángeles; Wright, Robert O.; Till, Christine; Cariology, Operative Dentistry and Dental Public Health, School of DentistryFoods and beverages provide a source of fluoride exposure in Mexico. While high fluoride concentrations are neurotoxic, recent research suggests that exposures within the optimal range may also pose a risk to the developing brain. This prospective study examined whether dietary fluoride intake during pregnancy is associated with toddlers' neurodevelopment in 103 mother-child pairs from the PROGRESS cohort in Mexico City. Food and beverage fluoride intake was assessed in trimesters 2 and 3 using a food frequency questionnaire and Mexican tables of fluoride content. We used the Bayley-III to evaluate cognitive, motor, and language outcomes at 12 and 24 months of age. Adjusted linear regression models were generated for each neurodevelopment assessment time point (12 and 24 months). Mixed-effects models were used to consider a repeated measurement approach. Interactions between maternal fluoride intake and child sex on neurodevelopmental outcomes were tested. Median (IQR) dietary fluoride intake during pregnancy was 1.01 mg/d (0.73, 1.32). Maternal fluoride intake was not associated with cognitive, language, or motor outcomes collapsing across boys and girls. However, child sex modified the association between maternal fluoride intake and cognitive outcome (p interaction term = 0.06). A 0.5 mg/day increase in overall dietary fluoride intake was associated with a 3.50-point lower cognitive outcome in 24-month old boys (95 % CI: -6.58, -0.42); there was no statistical association with girls (β = 0.07, 95 % CI: -2.37, 2.51), nor on the cognitive outcome at 12-months of age. Averaging across the 12- and 24-month cognitive outcomes using mixed-effects models revealed a similar association: a 0.5 mg/day increase in overall dietary fluoride intake was associated with a 3.46-point lower cognitive outcome in boys (95 % CI: -6.23, -0.70). These findings suggest that the development of nonverbal abilities in males may be more vulnerable to prenatal fluoride exposure than language or motor abilities, even at levels within the recommended intake range.Item Domain-specific effects of prenatal fluoride exposure on child IQ at 4, 5, and 6–12 years in the ELEMENT cohort(Elsevier, 2022) Goodman, Carly V.; Bashash, Morteza; Green, Rivka; Song, Peter; Peterson, Karen E.; Schnaas, Lourdes; Mercado-García, Adriana; Martínez-Medina, Sandra; Hernández-Avila, Mauricio; Martinez-Mier, Angeles; Téllez-Rojo, Martha M.; Hu, Howard; Till, Christine; Dental Public Health and Dental Informatics, School of DentistryObjective: Prenatal exposure to fluoride has been associated with adverse neurodevelopmental outcomes. However, the neuropsychological profile of fluoride's developmental neurotoxicity at low levels and the stability of this relationship across childhood has not been characterized. We investigated the longitudinal and domain specific effect of prenatal fluoride exposure on IQ among children ages 4, 5, and 6-12 years in the Early Life Exposures in Mexico to Environmental Toxicants (ELEMENT) cohort. Methods: We measured the average of maternal urinary fluoride at each trimester of pregnancy adjusted for creatinine (MUFCRE). Children were administered the McCarthy Scales of Children's Abilities at ages 4 (N = 386) and 5 (N = 308), and the Wechsler Abbreviated Scale of Intelligence at age 6-12 (N = 278). We used generalized estimating equation (GEE) models to estimate the population averaged effect of MUFCRE concentration on longitudinal General Cognitive Index (GCI)/Full-Scale IQ (FSIQ), Verbal IQ (VIQ), and Performance IQ (PIQ) scores (N = 348). We tested for possible interactions between MUFCRE and child sex as well as for MUFCRE and time point on children's IQ. All models controlled for relevant available covariates. Results: The mean/median MUFCRE concentration was 0.90/0.83 mg/L (SD = 0.39; IQR, 0.64-1.11 mg/L). A 0.5 mg/L increase in MUFCRE predicted an average 2.12-point decrease in GCI/FSIQ (95% CI: -3.49, -0.75) and 2.63-point decrease in PIQ (95% CI: -3.87, -1.40). MUFCRE was marginally associated with VIQ across time (B = -1.29, 95% CI: -2.60, 0.01). No interactions between MUFCRE and child sex or MUFCRE and time were observed. Conclusion: The negative association between prenatal fluoride exposure and longitudinal IQ was driven by decrements in non-verbal intelligence (i.e. PIQ), suggesting that visual-spatial and perceptual reasoning abilities may be more impacted by prenatal fluoride exposure as compared to verbal abilities.Item The effect of acid etching on remineralization of incipient caries lesions : a micro-ct study(2009) Yeslam, Hanin E.; Ando, Masatoshi; Gonzalez-Cabezas, Carlos, 1966-; Chu, Tien-Min Gabriel; Lund, Melvin; Cochran, MichaelEtching of enamel caries lesions has been demonstrated to enhance remineralization. However, this effect reaches a plateau after a period of time. This study aimed at investigating the effectiveness of additional acid etching on remineralization. Forty 1 mm × 2 mm human enamel blocks with chemically induced artificial incipient lesions were used. Ten specimens were randomly selected at the end of demineralization for transverse microradiography (TMR) analysis. The remaining specimens were then divided into three groups (n = 10). Group A was remineralized by a pH cycling system with 1100 ppm sodium fluoride for 20 days. In group B, the specimens were etched with 35-percent phosphoric acid for 30 s and then remineralized. Group C was remineralized by same procedure as group B plus and given an additional acid etch after 10 days of remineralization. Mineral density was measured by x-ray microtomography (µ-CT). The volumetric mineral content [VM (µm3×105)] was determined between 91 and 0-wt%. The µ-CT % mineral recovery (%) was calculated using the formula 100×(remineralize VM - demineralization VM) / (sound VM - demineralization VM). One-hundred-μm sections of demineralized and remineralized specimens were used to assess the mineral loss (IML: vol%×µm) and lesion depth (µm) using TMR. The three groups showed no significant difference in mineral change or mineral content for µ-CT or TMR lesion depth. The TMR IML showed a significant difference between the demineralized specimens and the three remineralized groups. The correlation between TMR IML and TMR lesion depth was 0.66 (p < 0.0001). The µ-CT percent mineral recovery from demineralization was correlated with neither TMR IML nor TMR lesion depth. When evaluated with µ-CT, the twice-acid-etched group presented lower mineral gain values than the group etched only once with acid. Also, the twice-etched group presented lower mineral gain and greater TMR IML compared with the non-acid etch group. TMR images revealed reduction of surface layer in the acid-etched groups, especially in the twice-etched group, in which significant reduction or loss of surface layer occurred. Based on these results, we conclude that additional acid etching with 35-percent phosphoric acid does not enhance remineralization compared with a single application of acid etching. We believe that the viable existence of the surface layer is essential for remineralization of the lesion. Further investigations into the accuracy of µ-CT to detect minute mineral changes in incipient caries lesions are probably needed.Item Effect of Bottled Water and Fluoride Toothpaste Usage on Caries Lesion Remineralization(2023) Qaw, Masoumah Samir; Lippert, Frank; Al Dehailan, Laila; Hara, Anderson T; Cook, N. BlaineBackground: The importance of fluoride in the prevention of dental caries has been well documented in the literature, as it inhibits demineralization of the tooth structure and enhances remineralization. One of the major public health policies to prevent caries is to provide the population with an adequate amount of fluoride through community water fluoridation. Nowadays many people drink bottled water instead of tap water due to its easy access, convenience, and low cost. Besides fluoride, other minerals present in tap and bottled water, such as calcium and magnesium, are also important in decreasing dental caries prevalence. However, our knowledge of the role of bottled water in caries prevention and especially when combined with fluoride toothpaste usage is still poor. Objectives: The aim of this in-vitro study was to evaluate the effects of some bottled waters on fluoride toothpaste efficacy in enhancing caries lesion remineralization. Methodology: Early caries lesions were created in bovine enamel specimens and stratified into treatment groups based on Vickers surface microhardness (VHN). The present study followed a two (fluoride and fluoride-free toothpaste) by five (four bottled waters and tap water) factorial design. The treatment groups were bottled water with the following attributes: a) 309.9 ppm Ca/1.20 ppm F; b) 118.4 ppm Ca/0.16 ppm F; c) 1.00 ppm Ca/1.01 ppm F; d) 0.1 ppm Ca/0.04 ppm F, and tap water (48.7 ppm Ca/0.7 ppm F). The five water groups were paired either with 1100 ppm fluoride or fluoride-free toothpaste, yielding 10 groups. Specimens were pH-cycled for 10 days with the daily regimen comprised of twice daily toothpaste slurry, with four exposures to water in between. VHN was measured, again, and the difference calculated (ΔVHN). Data were analyzed using two-way ANOVA at a 5-percent significance level. Results: The two-way interaction between water and toothpaste was significant (p < 0.001). All groups except fluoride-free toothpaste/bottled water with 0.1 ppm Ca/0.04 ppm F (p = 0.411) had significant increases in VHN after pH cycling (p ≤ 0.023). Fluoridated toothpaste resulted in a higher rate of remineralization compared to fluoridefree toothpaste (all p < 0.001). Bottled water with 1.20 ppm F/309.9 ppm Ca exhibited the greatest extent of remineralization within fluoride toothpaste groups (p < 0.001) and higher remineralization than lower fluoride water in fluoride-free toothpaste groups (p ≤ 0.006). Within the fluoridated toothpaste group, tap water exhibited significantly less remineralization compared to all bottled waters (all p < 0.001). Conclusion: Within the limitation of this study, bottled water with higher fluoride and calcium concentrations might improve fluoridated toothpaste efficacy by enhancing remineralization of early enamel caries-like lesions.Item Effects of a sodium fluoride- and phytate-containing dentifrice on remineralisation of enamel erosive lesions—an in situ randomised clinical study(Springer, 2018-02-08) Creeth, Jonathan E.; Parkinson, Charles R.; Burnett, Gary R.; Sanyal, Susmita; Lippert, Frank; Zero, Domenick T.; Hara, Anderson T.; Cariology, Operative Dentistry and Dental Public Health, School of DentistryObjectiveThe objective of this work was to evaluate effects of a dentifrice containing sodium fluoride (1150 ppm F) and the organic polyphosphate phytate (0.85% w/w of the hexa-sodium salt) on in situ remineralisation of early enamel erosive lesions and resistance to subsequent demineralisation.Materials and methodsSubjects (n = 62) wore palatal appliances holding eight bovine enamel specimens with pre-formed erosive lesions. They brushed their natural teeth with the phytate test dentifrice (TD); a positive control dentifrice (PC, 1150 ppm fluoride as NaF); a reference dentifrice (RD, disodium pyrophosphate + 1100 ppm fluoride as NaF) or a negative control dentifrice (NC, fluoride-free) in a randomised, double-blind, crossover design. Specimens were removed at 2, 4 and 8 h post-brushing and exposed to an ex vivo acid challenge. Surface microhardness (Knoop) was measured at each stage. The primary efficacy variable was relative erosion resistance (RER); other variables included the surface microhardness recovery (SMHR), acid resistance ratio (ARR) and enamel fluoride uptake (EFU).ResultsAfter 4 h, the results for RER, ARR and EFU were in the order PC > TD = RD > NC with PC > TD = RD = NC for SMHR. Results at 2 and 8 h were generally consistent with the 4 h data. Mineralisation progressed over time. Dentifrices were generally well-tolerated.ConclusionsIn this in situ model, addition of phytate or pyrophosphate to a fluoride dentifrice inhibited the remineralising effect of fluoride. Both formulations still delivered fluoride to the enamel and inhibited demineralisation, albeit to a lesser extent than a polyphosphate-free dentifrice.Clinical relevanceAddition of phytate or pyrophosphate to a fluoride dentifrice may reduce its net anti-erosive properties.Item Fluoride Content of Infant Formula Commercially Available in Central Indiana(2024) Altamimi, Ayman M.; Lippert, Frank; E. Soto-Rojas , Armando; Martinez Mier, E. AngelesBACKGROUND: Fluorides have a well-established role in dental caries prevention. Fluoride content in infant formula has raised concerns about whether it is within safe levels for the developing teeth. There is a large number of products on the market with likely varying fluoride concentrations, and these products’ fluoride content will differ depending on whether, for example, fluoridated water was used during manufacturing or reconstitution. Several studies have been published on infant formula containing fluoride and the associated risk of developing enamel fluorosis. However, few recent studies in the US have determined whether liquid or powder infant formula fall within safe/recommended levels. Purpose: This study measured the fluoride content of infant formula sold in grocery stores in central Indiana, prepared using three types of water (Purified, Nursery, and Tap) to determine if they fall within safe levels. Alternative hypotheses: There is a significant difference in the concentration of fluoride between different brands of infant formula. Material & Methods: We analyzed twenty different infant formula products sold in grocery stores in the Indianapolis, Indiana area for their fluoride content. Samples were reconstituted with Nursery water (containing approx. 1.0 ppm fluoride), Tap water (approx. 0.7 ppm fluoride) and Purified water (negligible fluoride content). A sample for the tests was taken from each preparation and the concentrations of fluoride of all samples was determined using the fluoride microdiffusion method. The statistical analysis of results was carried out using two-way ANOVA. Results: When comparing the mean (SD) fluoride concentration among the three types of infant formula reconstitution with water, tap water had significantly higher fluoride concentration mean than both Nursery water and purified water (P <.001 at α=.050 level). Nursery water also had significantly higher fluoride concentration mean than purified water (P <.001 at α=.050 level). When the three types of water were used for reconstitution of the 20 infant formula brands, the overall highest fluoride concentration mean was seen when tap water was used for reconstitution (0.950) followed by nursery water (0.789) while the least fluoride concentration was in purified water (0.102). Conclusion: Within the study's limitations, it can be concluded that apart from one formula none of the tested infant formulas sold in central Indiana grocery stores when reconstituted with purified water were found to decrease the chance of infants exceeding UL levels for both age groups but were found to increases the chance exceeding the AI levels for infants aged 0–6 months. All tested infant formulas reconstituted with nursery and tap water were found to increase the chance of infants exceeding the UL, and the AI levels for both groups resulted in increasing the chance of fluoride concentrations exceeding the recommended/safe levels. Thus, the type of water used for reconstitution rather than the type of formula appears to be the determining factor for the levels of fluoride intake associated with infant formula. Clinical Significance: With the recent increase in the utilization of infant formula, different brands with varying fluoride concentrations and the different modes of reconstitution must be evaluated to determine if their fluoride concentrations will fall within safe/recommended levels and thus increase the risk of enamel fluorosis development.
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