The in vitro remineralizing effect of CPP-ACP and CPP-ACPF after 6 and 12 weeks on initial caries lesion

Abstract Objective: The aim of this in vitro study was to determine the effects of remineralization promoting agents containing casein phosphopeptide-stabilized amorphous calcium phosphate (CPP-ACP), or CPP-ACP in combination with fluoride (CPP-ACPF) on artificial white spot lesions (WSLs) after 6 and 12 weeks. Methodology: White spot lesions were created on 123 sectioned premolars (246 specimens) with a demineralization solution during a 96 hours pH-cycling regime. Two experimental groups were created: a CPP-ACP group (Tooth Mousse™), and a CPP-ACPF group (Mi Paste Plus™). Additionally, two control groups were created, one using only a conventional toothpaste (1450 ppm fluoride) and another one without any working agents. All teeth were also daily brushed with the conventional toothpaste except the second control group. Tooth Mousse™ and Mi Paste Plus™ were applied for 180 seconds every day. The volume of demineralization was measured with transverse microradiography. Six lesion characteristics regarding the lesion depth and mineral content of WSLs were also determined. Results: The application of CPP-ACP and CPP-ACPF had a significant regenerative effect on the WSLs. Compared to Control group 1 and 2 the volume of demineralization after 6 weeks decreased significantly for CPP-ACP (respectively p<0.001 and p<0.001) and CPP-ACPF (respectively p=0.001 and p=0.003). The same trend was observed after 12 weeks. For the CPP-ACPF group, WSL dimensions decreased significantly between 6 and 12 weeks follow-up (p=0.012). The lesion depth reduced significantly after application of CPP-ACP and CPP-ACPF but increased significantly in the Control groups. Mineral content increased for CPP-ACP and CPP-ACPF after an application period of 12 weeks, but this was only significant for CPP-ACP. Conclusions: Long-term use of CPP-ACP and CPP-ACPF in combination with a conventional tooth paste shows beneficial effects in the recovery of in vitro subsurface caries lesions.

Remineralizing effect of commercial fluoride varnishes on artificial enamel lesions

Abstract: The aim of this study was to evaluate soluble and insoluble fluoride concentrations in commercial varnishes, and their remineralization effect on artificial caries enamel lesions using surface and cross-sectional microhardness evaluations. Forty bovine enamel blocks were separated into four groups (n=10): control (no treatment), Enamelast (Ultradent Products), Duraphat (Colgate-Palmolive) and Clinpro White Varnish (3M ESPE). Surface enamel microhardness evaluations were obtained, artificial enamel caries lesions were developed by dynamic pH-cycling, and the varnishes were then applied every 6 days, after which the enamel blocks were submitted to dynamic remineralization by pH cycles. After removal of the varnishes, the enamel surfaces were reassessed for microhardness. The blocks were sectioned longitudinally, and cross-sectional microhardness measurements were performed at different surface depths (up to 300 μm depth). Polarized light microscopy images (PLMI) were made to analyze subsurface caries lesions. The fluoride concentration in whole (soluble and insoluble fluoride) and centrifuged (soluble fluoride) varnishes was determined using an extraction method with acetone. The data were analyzed to evaluate the surface microhardness, making adjustments for generalized linear models. There was a significant decrease in enamel surface microhardness after performing all the treatments (p<0.0001). Enamelast and Duraphat showed significantly higher enamel microhardness values than the control and the Clinpro groups (p = 0.0002). Microhardness loss percentage was significantly lower for Enamelast (p = 0.071; One-way ANOVA). PLMI showed that subsurface caries lesions were not remineralized with the varnish treatments. No significant differences in the in-depth microhardness levels (p = 0.7536; ANOVA) were observed among the treatments. Enamelast presented higher soluble and insoluble fluoride concentrations than the other varnishes (p < 0.0001; Kruskal-Wallis and Dunn tests). Enamelast and Duraphat varnishes promoted enamel surface remineralization, but no varnish remineralized the subsurface lesion body. Although insoluble and soluble fluoride concentration values did not correspond to those declared by the manufacturer, Enamelast presented higher fluoride concentration than the others.

Effect of a resin-modified glass-ionomer with calcium on enamel demineralization inhibition: an in vitro study

Abstract We assessed the effect of a new coating material based on resin-modified glass-ionomer with calcium (Ca) in inhibiting the demineralization of underlying and adjacent areas surrounding caries-like lesions in enamel. The measures used were surface hardness (SH) and cross-sectional hardness (CSH). Thirty-six bovine enamel specimens (3 × 6 × 2 mm) were randomly allocated into three groups (n = 12): No treatment (NT); resin-modified glass-ionomer with Ca (Clinpro XT Varnish, 3M ESPE) (CL), and fluoride varnish (Duraphat, Colgate) (DU). The specimens were subjected to alternated immersions in demineralizing (6 h) and remineralizing solutions (18 h) for 7 days. SH measurements were conducted at standard distances of 150, 300, and 450 µm from the treatment area. CSH evaluated the mean hardness profile over the depth of the enamel surface and at standard distances from the materials. The energy-dispersive X-ray spectroscopy analysis was conducted to evaluate the demineralization bands created on the sublayer by % of calcium (Ca), phosphorus (P), and fluoride (F). Ca/P weight ratio was also calculated. Based on SH and CSH measurements, there was no difference between groups at the distances 150 µm (p = 0.882), 300 µm (p = 0.995), and 450 µm (p = 0.998). Up to 50 µm depth (at 150 µm from the treatment area), CL showed better performance than DU ( p< 0.05). NT presented higher loss of Ca and P than CL and DU (p < 0.05). There was no significant difference in the % of F ion among the three groups. The new coating material was similar to F varnish in attenuating enamel demineralization.

Effect of fluoride application during radiotherapy on enamel demineralization

Abstract Radiation-related caries are one the most undesired reactions manifested during or after head and neck radiotherapy. Fluoride application is an important strategy to reduce demineralization and enhance remineralizaton. Objective: To evaluate the effect of the topical application of fluoride during irradiation on dental enamel demineralization. Material and Methods: Thirty molars were randomly divided into three groups: Non-irradiated (NI), Irradiated (I), Irradiated with fluoride (IF). Each group was subdivided according to the presence or absence of pH-cycling (n=5). In the irradiated groups, the teeth received 70 Gy. The enamel's chemical composition was measured using Fourier Transform Infrared Spectrometry (organic matrix/mineral ratio - M/M and relative carbonate content - RCC). Vickers microhardness (VHN) and elastic modulus (E) were evaluated at three depths (surface, middle and deep enamel). Scanning electron microscopy (SEM) was used to assess the enamel's morphology. Results: The FTIR analysis (M/M and RCC) showed significant differences for irradiation, pH-cycling and the interaction between factors (p<0.001). Without pH-cycling, IF had the lowest organic matrix/mineral ratio and relative carbonate content. With pH-cycling, the organic matrix/mineral ratio increased and the relative carbonate content decreased, except for IF. VHN was influenced only by pH-cycling (p<0.001), which generated higher VHN values. ANOVA detected significant differences in E for irradiation (p<0.001), pH-cycling (p<0.001) and for the interaction between irradiation and pH-cycling (p<0.001). Increased E was found for group I without pH-cycling. With pH-cycling, groups I and IF were similar, and showed higher values than NI. The SEM images showed no morphological changes without pH-cycling. With pH-cycling, fluoride helped to maintain the outer enamel's morphology. Conclusions: Fluoride reduced mineral loss and maintained the outer morphology of irradiated and cycled enamel. However, it was not as effective in preserving the mechanical properties of enamel. Radiotherapy altered the enamel's elastic modulus and its chemical composition.

Caries lesion remineralization with fluoride toothpastes and chlorhexidine - effects of application timing and toothpaste surfactant

Abstract Habitual toothbrushing with fluoridated toothpaste followed by rinsing with antibacterial mouthwashes is a method to maintain good oral hygiene and to diminish the occurrence and severity of dental caries and periodontal disease. However, our understanding of how antimicrobial agents in mouthwashes affect fluoride-mediated caries lesion remineralization is still poor. Objective: The objectives of this in vitro study were a) to determine the effects of the waiting period of chlorhexidine (CHX) rinsing after fluoride toothpaste use and b) to further determine the effect of the type of toothpaste surfactant [sodium dodecyl sulfate (SDS) or cocamidopropyl betaine (CAPB)] on caries lesion remineralization associated with CHX rinsing. Material and Methods: Caries lesions were formed in bovine enamel specimens and assigned to 10 treatment groups (n=18) based on Vickers surface microhardness (VHN). Lesions were then pH-cycled for 10 days with daily regimen comprised of twice daily toothpaste slurry treatments (1150 ppm fluoride, with SDS or CAPB), followed by CHX solution treatments [0, 15, 30 or 60 minutes following slurry treatment or no CHX treatment (negative control)]. VHN was measured again and the extent of lesion remineralization calculated (∆VHN). Results: ∆VHN with SDS-toothpaste was significantly lower than with CAPB-toothpaste, indicating more remineralization for the CAPB-toothpaste. ∆VHN with 0-minute waiting time was significantly lower than with 30-minute waiting time and with negative control. Conclusions: The absence of CHX as an adjunct to fluoride toothpastes led to greater remineralization of enamel lesions compared with the immediate use of CHX treatment for both SDS- and CAPB-toothpastes. CAPB-toothpastes indicated significantly greater remineralization than SDS-toothpastes, and can be suggested for patients at high risk of caries. A 30-minute waiting time for CHX treatment is recommended after brushing.

TiF4 varnish protects the retention of brackets to enamel after in vitro mild erosive challenge

Abstract The effect of fluoride agents on the retention of orthodontic brackets to enamel under erosive challenge is little investigated. Objective: The aim of this study was to evaluate the effect of titanium tetrafluoride (TiF4) and sodium fluoride (NaF) agents on the shear bond strength of brackets to enamel and on the enamel microhardness around brackets under erosive challenge. Methods: Brackets were bonded to bovine incisors. Five groups were formed according to fluoride application (n=10): TiF4 varnish, TiF4 solution, NaF varnish, NaF solution and control (without application). The specimens were submitted to erosive challenge (90 s cola drink/2h artificial saliva, 4x per day for 7 days). Solutions were applied before each erosive cycle and varnishes were applied once. Vickers Microhardness (VHN) was obtained before and after all cycles of erosion and the percentage of microhardness loss was calculated. Shear bond strength, adhesive remnant index and polarized light microscopy were conducted after erosion. The data were analyzed by ANOVA, Tukey, Kruskal-Wallis and Mann-Whitney U tests (α=0.05). Results: The %VHN had no statistically significant differences among the experimental groups. However, considering the comparisons of all groups with the control group, TiF4 varnish showed the highest protection from enamel demineralization (effect size of 2.94, while the effect size for the other groups was >2.4). The TiF4 varnish group had significantly higher shear bond strength compared to other groups. There was no difference among groups for adhesive remnant index. Polarized light microscopy showed higher demineralization depth for the control group. Conclusions: Application of NaF and TiF4 agents during mild erosive challenge minimized the enamel mineral loss around brackets, however only the experimental TiF4 varnish was able to prevent the reduction of shear bond strength of brackets to enamel.

Effect of fluoride on salivary immunoglobulins and sialic acid

Summary Objective: The aim of our study was to evaluate the effect of fluoride on salivary immunoglobulin and sialic acid levels in children with dental fluorosis and healthy teeth who live in places with high fluoride concentration in drinking water. Method: Fifty-one (51) healthy children between 6 and 12 years old with no caries were randomly selected from primary schools enrolled in the dental-care program operated by the Department of Pediatric Dentistry. The children were divided into two groups: group I comprised 26 children with dental fluorosis [Thylstrup-Fejerskov Dental Fluorosis Index (TFI) = 4] who lived in Isparta (2.7-2.8 ppm), and group II consisted of 25 children without dental fluorosis who were born in low-fluoride areas and had lived in Isparta for only the previous two years. Stimulated and unstimulated saliva were collected and analyzed for fluoride, salivary immunoglobulins and sialic acid levels. Results: Sialic acid level was correlated negatively with age. Levels of secretory immunoglobulin A (sIgA) and secretory immunoglobulin G (sIgG) were higher in children with dental fluorosis compared with those in group II, although these differences were not significant. Conclusion: Increased sIgA and sIgG levels may arrest the progression of caries in subjects with dental fluorosis. Given the risks of dental fluorosis, further studies of the effects of different fluoride levels in drinking water on salivary composition of children with mixed dentition are needed to confirm the results of our study and to provide data for comparison.

The use of sodium trimetaphosphate for matrix metalloproteinase inhibition, remineralization and bonding to dentin / O uso de trimetafosfato de sódio sobre a inibição de metaloproteinases e na remineralização e adesão à dentina

The adhesive process to dentin substrate depends on the condition determined by the combined action of the mineral loss and the endogenous enzymes activity. Thus, considering a more complete therapeutic approach, sodium trimetaphosphate (STMP) may be a novel strategy that conciliates the remineralization potential to the promotion of dentin strengthening and its stability, possibly directing mineral nucleation and controlling the rate of biodegradation. In this study, the effect of STMP was evaluated in 2 studies. In study 1, different concentrations of STMP (0.5, 1.5, 3.5 and 5%) were investigated to assess their anti-proteolytic capacity on human purified MMPs-2 and -9 by zymography. Afterwards, only the concentrations (1.5, 3.5 and 5%) that showed total inhibition of both MMPs were used to evaluate their remineralizing capacity in dentin substrate submitted to artificial cariogenic challenge, through surface hardness (SH) and cross-sectional hardness (CSH). In study 2, based on the previous results, the capacity of the 1.5% STMP associated or not with NaF or Ca(OH)2 solutions in improving the dentin bond strength of a universal adhesive system was evaluated by the microtensile test . Thus, these studies suggest that 1.5% STMP is an effective inhibitor of collagen degradation mediated by purified human MMPs-2 and -9. In addition, demineralized and treated dentin with 1.5% STMP supplemented with Ca(OH)2 may induce remineralization. Thus, the use of STMP can be introduced as a new strategy that combines enzymatic inhibition and remineralization potential, reestablishing favorable conditions to affected dentin. These evidences support perspectives of therapies to restructure dentin and propose feasible and promising clinical strategies.(AU)

O processo adesivo ao substrato dentinário depende da condição determinada pela ação combinada da perda mineral e atividade de enzimas endógenas. Deste modo, considerando uma abordagem terapêutica mais completa, o trimetafosfato de sódio (STMP) pode ser uma estratégia inovadora que concilia o potencial remineralizador à promoção do fortalecimento da dentina e sua estabilidade, possivelmente direcionando a nucleação mineral e controlando a taxa de biodegradação. Neste trabalho, o efeito do STMP foi avaliado em 2 estudos. No estudo 1, diferentes concentrações de STMP (0,5; 1,5; 3,5 e 5%) foram investigadas para avaliar sua capacidade anti-proteolítica sobre as MMPs-2 e -9 purificadas humanas, por zimografia. Posteriormente, somente as concentrações (1,5; 3,5 e 5%) que apresentaram capacidade de inibição total de ambas MMPs foram utilizadas para avaliar sua capacidade remineralizadora em substrato dentinário submetido ao desafio cariogênico artificial, através da dureza de superfície (DS) e longitudinal (DL). No estudo 2, baseado nos resultados anteriores, foi avaliada a capacidade do STMP à 1,5% associado ou não a soluções de NaF ou Ca(OH)2 em melhorar a resistência de união à dentina de um sistema adesivo universal pelo teste de microtração. Desta forma, estes estudos sugerem que o STMP à 1,5% apresenta-se como um inibidor eficaz da degradação do colágeno mediada por MMPs-2 e -9 humanas purificadas. Além disso, a dentina humana desmineralizada e tratada com STMP à 1,5% suplementada com Ca(OH)2 pode induzir à remineralização. Assim, o uso de STMP pode ser introduzido como uma nova estratégia que combina inibição enzimática e potencial de remineralização, reestabelecendo condições favoráveis a partir de uma dentina afetada. Estas evidências sustentam perspectivas de terapias para reestruturar a dentina e propor estratégias clínicas factíveis e promissoras.(AU)

Protective effect of milk against dental demineralization: understanding the mechanisms / Efeito protetor do leite contra a desmineralização dentária: compreensão dos mecanismos

This study aimed to answer the following questions: 1) does whole fluoridated milk protect more against enamel and dentin erosion than fat-free fluoridated milk? 2) does the protective effect of fluoridated milk against erosion follow a dose-response relationship? 3) is the treatment with whole or fat-free fluoridated milk before the first erosive challenge more protective against enamel and dentin erosion? 4) does the fat content of milk change the proteomic profile of the acquired enamel pellicle (AEP)? This study was divided into 2 parts. The first part analyzed in vitro the effect of milk against dental erosion, considering three factors: type of bovine milk (whole/fat-free), presence of different fluoride concentrations (0- 10.0 ppm) and time of application (before/after erosive challenge). Bovine enamel (n=15/group) and root dentin (n=12/group) specimens were submitted to the following treatments: 0.9% NaCl solution (negative control)( after first erosive challenge); whole milk with 0, 2.5, 5.0, 10.0 ppm F; fat-free milk with 0, 2.5, 5.0, 10.0 ppm F; 0.05% NaF solution (positive control) (before or after first erosive challenge). Specimens were submitted to demineralization - remineralization regimes, 4 times/ day, for 5 days. The response variables were enamel and dentin loss, evaluated by profilometry (µm). Data were analyzed using Kruskal­Wallis/Dunn's test (p<0.05). The presence of fluoride, especially at 10 ppm, was the most important factor in reducing dental erosion. The second part detected changes in protein profile of AEP formed in vivo after rinsing with whole milk, fat-free milk or water. Nine subjects with good oral conditions participated. The AEP was formed in the morning, for 120 min, after prophylaxis with pumice. In sequence, the volunteers rinsed with 10 mL of whole milk, fat-free milk or deionized water for 30 s, following a blind, crossover protocol. After 60 min, the AEP was collected with filter paper soaked in 3% citric acid and processed for analysis by liquid chromatography-electrospray ionization tandem mass spectrometry (LCESI- MS/MS). The obtained MS/MS spectra were searched against human protein database (SWISS­PROT). The proteomic data related to protein quantification were analyzed using the PLGS software. A total of 260 proteins were successfully identified in the AEP samples collected in all groups. Forty-nine were common to the 3 groups, while 72, 62 and 49 were specific for groups treated with whole milk, fat-free milk and water, respectively. Some were typical components of the AEP, such as Cystatin-B, Lysozyme C, Histatin-1, Statherin and Lactotransferrin. Other proteins are not commonly described as part of the AEP but could act in the defense of the organism against pathogens. Distinct proteomic profiles were found in the AEP after rinsing with whole or fat-free milk, which could have an impact in bacterial adhesion and tooth dissolution. The use of fat-free milk could favorably modulate the adhesion of bacteria in the AEP and the biofilm formation in comparison to whole milk.(AU)

Este estudo objetivou responder as seguintes questões: 1) o leite integral fluoretado protege mais contra a erosão do esmalte e dentina do que o leite fluoretado desnatado? 2) o efeito protetor do leite fluoretado segue um padrão dose-resposta? 3) o tratamento com leite integral ou leite desnatado fluoretado antes do primeiro desafio erosivo protege mais contra a erosão do esmalte e dentina? 4) o leite contendo gordura altera o perfil proteico da película adquirida do esmalte (PAE)? O estudo foi dividido em 2 partes. Na primeira parte foi realizado um estudo in vitro, considerando três fatores: tipo de leite bovino (integral/ desnatado), diferentes concentrações de fluoreto e tempo de aplicação (antes/após desafio erosivo). Os espécimes de esmalte bovino (n=15 /grupo) e dentina radicular (n=12 /grupo) foram submetidos aos seguintes tratamentos: solução de NaCl a 0,9% (controle negativo)(após o desafio erosivo); Leite integral com 0, 2,5, 5,0, 10,0 ppm F Leite desnatado com 0, 2,5, 5,0, 10,0 ppm F 0,05% de solução de NaF (controle positivo) (antes ou após o primeiro desafio erosivo). Os espécimes foram submetidos a regimes de desmineralização e remineralização, 4 vezes/dia, durante 5 dias. As variáveis de resposta foram perda de esmalte e dentina, avaliadas por perfilometria (µm). Os dados foram analisados usando o teste de Kruskal-Wallis / Dunn (p <0,05). A presença de fluoreto, especialmente na concentração de 10 ppm, demonstrou ser o fator mais importante na redução da erosão dentária. A parte II do estudo detectou alterações no perfil proteico da PAE formada in vivo após bochecho com leite integral, leite desnatado ou água. Nove indivíduos com boas condições de saúde bucal participaram. A PAE foi formada pela manhã, durante 120 minutos, após profilaxia com pedra-pomes. Em seguida, os voluntários bochecharam com 10 mL de leite integral, leite desnatado ou água deionizada durante 30 s, seguindo um protocolo cego e cruzado. Após 60 min, a película foi coletada com papel de filtro embebido em ácido cítrico a 3% e processada para análise por cromatografia líquida acoplada à espectrometria de massas com ionização por eletrospray (LC-ESI-MS / MS). Os espectros MS/MS obtidos foram confrontados com bases de dados de proteínas humanas (SWISSPROT). Os dados proteômicos relacionados à quantificação de proteínas foram analisados usando o software PLGS. Um total de 260 proteínas foi identificado nas amostras de PAE coletadas em todos os grupos. Quarenta e nove eram comuns aos 3 grupos, enquanto 72, 62 e 49 eram específicas para grupos tratados com leite integral, leite desnatado e água, respectivamente. Algumas proteínas encontradas são típicas da PAE, como Cistatina-B, Lisozima C, Histatina-1, Estaterina e Lactotransferrina. Outras proteínas não são comumente descritas como parte da PAE, mas podem atuar na defesa do organismo contra patógenos. Perfis proteômicos distintos foram encontrados na PAE após o bochecho com leite integral ou desnatado, o que poderia ter um impacto na adesão bacteriana e na dissolução dentária. O uso de leite desnatado pode modular favoravelmente a adesão de bactérias na PAE e a formação do biofilme em comparação com o leite integral.(AU)

Influence of biofilm formation on the mechanical properties of enamel after treatment with CPP-ACP crème

Abstract: The study aimed to investigate the effects of bacterial biofilms on changes in the surface microhardness of enamel treated with casein phosphopeptide—amorphous calcium phosphate (CPP-ACP) with and without fluoride. Human enamel blocks with incipient caries-like lesions were divided into four groups of 13: G1: Saliva (Control); G2: fluoride dentifrice (Crest™, 1100 ppm as NaF); G3: CPP-ACP (MI Paste; Recaldent™); and G4: CPP-ACPF (MI Paste Plus; Recaldent™ 900 ppm as NaF). The specimens were soaked in demineralizing solution for 6 h and remineralized in artificial saliva for 18 h alternately for 10 days. The dentifrice was prepared with deionized water in a 1 : 3 ratio (w/w) or applied undiluted in the case of the CPP-ACP group. The surface microhardness (SMH) was evaluated at baseline, after artificial caries, after pH cycling and treatment with dentifrices, and after incubation in media with Streptococcus mutans for biofilm formation. The biofilms were exposed once a day to 2% sucrose and the biofilm viability was measured by MTT reduction. The percentage of change in surface microhardness (%SMHC) was calculated for each block. The data were analyzed by nonparametric test comparisons (α = 0.05). The %SMHC values observed in G2 were different from those of G1, G3, and G4 (p < 0.05). After biofilm formation, %SMHC was positive in G2 and G4 when compared to G1 and G3, but resistance to demineralization after biofilm formation was similar in all groups. In conclusion, the presence of biofilms did not influence the treatment outcomes of anticaries products.

Evaluation of the remineralization capacity of CPP-ACP containing fluoride varnish by different quantitative methods

ABSTRACT Objective The aim of this study was to evaluate the efficacy of CPP-ACP containing fluoride varnish for remineralizing white spot lesions (WSLs) with four different quantitative methods. Material and Methods Four windows (3x3 mm) were created on the enamel surfaces of bovine incisor teeth. A control window was covered with nail varnish, and WSLs were created on the other windows (after demineralization, first week and fourth week) in acidified gel system. The test material (MI Varnish) was applied on the demineralized areas, and the treated enamel samples were stored in artificial saliva. At the fourth week, the enamel surfaces were tested by surface microhardness (SMH), quantitative light-induced fluorescence-digital (QLF-D), energy-dispersive spectroscopy (EDS) and laser fluorescence (LF pen). The data were statistically analyzed (α=0.05). Results While the LF pen measurements showed significant differences at baseline, after demineralization, and after the one-week remineralization period (p<0.05), the difference between the 1- and 4-week was not significant (p>0.05). With regards to the SMH and QLF-D analyses, statistically significant differences were found among all the phases (p<0.05). After the 1- and 4-week treatment periods, the calcium (Ca) and phosphate (P) concentrations and Ca/P ratio were higher compared to those of the demineralization surfaces (p<0.05). Conclusion CPP-ACP containing fluoride varnish provides remineralization of WSLs after a single application and seems suitable for clinical use.

Comparação de diferentes pré-tratamentos de superfície na remineralização de esmalte com flúor em tratamento de lesões com mancha branca e sua resistência após desafio ácido / Comparison of different pretreatments on remineralization of enamel white spot lesions with fluoride and its resistance after new acidic challenge

A aplicação de flúor tópico é a principal estratégia de natureza química para a remineralização de lesões incipientes (LI) clinicamente visíveis como manchas brancas (MB) por cárie. Abordagens para aumentar a retenção de F no substrato pode favorecer sua ação e para isso, tratamentos prévios da superfície do esmalte podem ser usados. O objetivo deste trabalho foi comparar a capacidade de remineralização do flúor sem e com pré-tratamento do esmalte com ácido fosfórico e nitrato de alumínio e a sua resistência após novo desafio. Sessenta espécimes de esmalte bovino foram preparados (6mm x 4mm) e selecionados por meio de análise de microdureza de superfície (MS). LI foram produzidas através de ciclagem Desmineralização-Remineralização (DES-RE) e os espécimes divididos aleatoriamente em cinco grupos, de acordo com o tratamento (n=12): V- controle (verniz de fluoreto de sódio 5% por 4 horas), F (sem pré-tratamento); P-F(condicionamento ácido com ácido fosfórico por 30s); Al-F (nitrato de alumínio a 0,05M por 1min); P-Al-F (condicionamento com ácido fosfórico a 37% + nitrato de alumínio a 0,05M). Os tratamentos foram repetidos semanalmente durante quatro semanas. Após o tratamento, os espécimes foram submetidos à nova ciclagem ácida. Após cada etapa, nova MS foi realizada e ao final das análises, um corte transversal dos espécimes foi realizado. Uma das metades foi submetida à análise da microdureza longitudinal (ML) e a outra preparada para realização da microrradiografia transversal (TMR) para a análise de conteúdo mineral perdido. O percentual de perda de dureza de superfície (%PDS) foi analisado por ANOVA a 2 critérios de medidas repetidas e Tukey e o percentual de perda de dureza longitudinal (%PDL) por ANOVA a 3 critérios de medidas repetidas e Tukey (p<0,05). Os dados de TMR no parâmetro LD (profundidade da lesão) foram analisados por ANOVA a dois critérios (p<0,05). Os resultados mostraram que a ciclagem DES-RE resultou em significante %PDS e %PDL em todos os grupos. Grupo F revelou menor perda de MS após tratamento e, F, Al-F e P-Al-F, mostraram menor perda de dureza final, após o novo desafio ácido. Na análise de %PDL, o grupo V apresentou menor perda de dureza final quando comparado com os demais grupos, nas diferentes profundidades. A análise do conteúdo mineral não revelou nenhuma diferença entre os tratamentos e fases. Nenhum dos pré tratamentos propostos foram capazes de otimizar a atuação do gel APF na remineralização de MB.(AU)

The topical fluoride is the main chemical strategy to remineralize incipient caries lesions (ICL) visible as carious white spot lesions (WSL). Approaches to increase the fluoride retention may favor its action and therefore, enamel pretreatments can be used for this purpose. The aim of this study was to compare the potential of fluoride remineralization with and without previous enamel pretreatment with aluminum nitrate and phosphoric acid as well as their resistance after a new acid challenge. Sixty bovine enamel specimens were prepared (6mm x 4mm) and selected by the surface hardness (SH) analysis. The ICL were produced using DE-RE cycling and the specimens were randomized in five groups, according to the treatment (n=12): Vcontrol (5% sodium fluoride varnish during 4h), and four groups previously preatreated with topical application of acidulated phosphate fluoride (APF) during 4min: F (without pretreatment), P-F (phosphoric acid etching during 30s), Al-F (0.05M aluminum nitrate during 1min); P-Al-F (phosphoric acid etching + aluminum nitrate). The treatments were performed weekly during four weeks. After the treatment, the specimens were submitted to the new acid challenge. After each step, a new SH analysis was performed followed by the transversal cut of the specimens. A half was submitted to the longitudinal cross-sectional hardness analysis (LH) and the other half was prepared to transverse microradiography assessments (TMR) to determine the loss of mineral content. The percentage of surface mineral loss (%SH) was analyzed using two-way repeated-measures ANOVA and the percentage of crosssectional mineral loss (%LH) by the three-way repeated-measures ANOVA and Tukey (p<0.05). Data of TMR analysis by LD (lesion depth) parameter was analyzed with two-way ANOVA (p<0.05). The results showed significant %SH and %LH after DE-RE cycling for all groups. F group showed the lowest %SH after treatment, with no significant difference to Al-F and P-Al-F after the new acid challenge. V group showed the lowest %LH compared to the other groups, in the different depths, suggesting more resistance. Mineral content assessment did not reveal any difference among treatments and phases. None pretreatments were able to increase the potential of fluoride remineralization.(AU)

Fluoride and sodium trimetaphosphate (TMP) release from fluoride varnishes supplemented with TMP

Abstract This study assessed fluoride (F) and sodium trimetaphosphate (TMP) release into artificial saliva from varnishes containing 0%, 2.5%, and 5% NaF, supplemented or not with 5% TMP. The varnishes were applied on polyester sheets (n = 8/group), and F and TMP released into artificial saliva were measured for up to 24 hours. The amount of F and TMP released were directly related to NaF and TMP concentrations in the varnishes. The highest F release was seen for 5% NaF and 5% NaF + 5% TMP, whereas 5% TMP released the highest amount of TMP. However, the simultaneous addition of NaF and TMP to varnishes significantly reduced the amount of F and TMP released from the products.

Thickness and nanomechanical properties of protective layer formed by TiF4 varnish on enamel after erosion

Abstract The layer formed by fluoride compounds on tooth surface is important to protect the underlying enamel from erosion. However, there is no investigation into the properties of protective layer formed by NaF and TiF4 varnishes on eroded enamel. This study aimed to evaluate the thickness, topography, nanohardness, and elastic modulus of the protective layer formed by NaF and TiF4 varnishes on enamel after erosion using nanoindentation and atomic force microscopy (AFM). Human enamel specimens were sorted into control, NaF, and TiF4 varnish groups (n = 10). The initial nanohardness and elastic modulus values were obtained and varnishes were applied to the enamel and submitted to erosive challenge (10 cycles: 5 s cola drink/5 s artificial saliva). Thereafter, nanohardness and elastic modulus were measured. Both topography and thickness were evaluated by AFM. The data were subjected to ANOVA, Tukey's test and Student's t test (α = 0.05). After erosion, TiF4 showed a thicker protective layer compared to the NaF group and nanohardness and elastic modulus values were significantly lower than those of the control group. It was not possible to measure nanohardness and elastic modulus in the NaF group due to the thin protective layer formed. AFM showed globular deposits, which completely covered the eroded surface in the TiF4 group. After erosive challenge, the protective layer formed by TiF4 varnish showed significant properties and it was thicker than the layer formed by NaF varnish.

Protective effect of experimental mouthrinses containing NaF and TiF4 on dentin erosive loss in vitro

Objective This in vitro study assessed the anti-erosive effect of experimental mouthrinses containing TiF4 and NaF on dentin erosive loss.Material and Methods Bovine dentin specimens were randomly allocated into the groups (n=15): 1) SnCl2/NaF/AmF (Erosion Protection®/GABA, pH 4.5, positive control); 2) experimental solution with 0.0815% TiF4(pH 2.5); 3) 0.105% NaF (pH 4.5); 4) 0.042% NaF+0.049% TiF4 (pH 4.4); 5) 0.063% NaF+0.036% TiF4 (pH 4.5); 6) no treatment (negative control). Each specimen was cyclically demineralized (Sprite Zero, pH 2.6, 4x90 s/day) and exposed to artificial saliva between the erosive challenges for 7 days. The treatment with the fluoride solutions was done 2x60 s/day, immediately after the first and the last erosive challenges of the day. Dentin erosive loss was measured by profilometry (μm). The data were analyzed using Kruskal Wallis/Dunn tests (p<0.05).Results Mouthrinses containing TiF4or Sn/F were able to show some protective effect against dentin erosive loss compared to negative control. The best anti-erosive effect was found for experimental solution containing 0.0815% TiF4 (100% reduction in dentin loss), followed by 0.042% NaF+0.049% TiF4 (58.3%), SnCl2/NaF/AmF (52%) and 0.063% NaF+0.036% TiF4 (40%). NaF solution (13.3%) did not significantly differ from control.Conclusion The daily application of experimental mouthrinse containing TiF4and NaF has the ability to reduce dentin erosion, as well as Erosion Protection® and TiF4 alone.

Avaliação da interação do TiF4 com a hidroxiapatita e esmalte bem como o efeito sobre a des-remineralização do esmalte dentário com lesão de cárie artificial in situ / Interaction of TiF4 varnish with enamel and hydroxyapatite and its effect on the de-remineralization of pre-demineralized enamel in situ

Este estudo teve como o objetivo avaliar in vitro o efeito do tratamento com vernizes de tetrafluoreto de titânio (TiF4) e fluoreto de sódio (NaF) (ambos com 0,95%, 1,95% e 2,45% F) sobre 1) a quantidade de F- liberada em água deionizada e saliva artificial, por período de 12h; 2) os tipos de compostos formados pela interação com a hidroxiapatita (neste caso, com soluções ao invés dos vernizes); 3) a porcentagem de elementos presentes na superfície do esmalte bovino e humano, hígidos (H) e desmineralizados (DES); 4) a quantidade de CaF2 sobre a superfície do esmalte bovino e humano, H e DES. Adicionalmente, 5) o efeito dos vernizes de TiF4 e NaF sobre a remineralização do esmalte bovino, em diferentes condições in situ, foi avaliado. Para etapa 1, a liberação de F- foi quantificada por eletrodo de íon específico. Na etapa 2, pó de HAP produzido por precipitação foi tratado com soluções fluoretadas. Os compostos formados foram avaliados por espectroscopia de infravermelho e difração de raios-X. Nas etapas 3 e 4, as superfícies tratadas com os vernizes foram analisadas por MEV- EDAX e por biópsias básicas, respectivamente. Vinte voluntários (n final=17) participaram do ensaio in situ com 3 fases cruzadas, os quais utilizaram aparelhos palatinos contendo amostras de esmalte bovino desmineralizadas tratadas com vernizes de TiF4, NaF ou placebo. As amostras foram submetidas a diferentes condições de des-remineralização (presença ou não de tela plástica; variação da frequência de aplicação de sacarose 20%) e à exposição ao dentifrício fluoretado. O volume mineral e profundidade da lesão foram avaliados por microradiografia transversal (TMR). 1) Os vernizes de TiF4 (1,95 e 2,45% F) liberaram mais fluoreto comparados aos vernizes de NaF tanto em água como em saliva artificial, sendo a diferença significativa nas primeiras 6h de contato (p<0,0001). 2) O TiF4 causou alteração na estrutura da HAP e induziu à formação de novos compostos como TiO2 e Ti (HPO4)2. 3)...

This study aimed to evaluate the in vitro effect of titanium tetrafluoride (TiF4) and sodium fluoride (NaF) varnishes (both with 0.95%, 1.95%, 2.45% F) treatment on the 1) F- release in deionized water and artificial saliva for 12h period; 2) types of compounds formed by the interaction with hydroxyapatite (in this case, solutions were tested instead of varnishes); 3) percentage of the elements on bovine and human, sound (S) and demineralized (DE), enamel surface; 4) F- uptake (CaF2 deposition) on human and bovine, S and DE, enamel surface. Additionally, 5) the effect of TiF4 and NaF varnishes on bovine enamel remineralization, in different in situ conditions, was assessed. In study 1, the F- release was measured by ion specific electrode. In study 2, HAP powder, produced by precipitation, was treated with fluoride solutions. The compounds formed were evaluated by infrared spectroscopy and X-ray diffraction. In studies 3 and 4, the surfaces treated with the varnishes were analyzed by SEM-EDAX and basic biopsies, respectively. Twenty volunteers (final n=17) participated of the in situ study with 3 crossover phases, in which they wore palatal appliances containing bovine demineralized enamel samples treated with TiF4, NaF or placebo varnishes. The samples were subjected to different de-remineralization conditions (presence or absence of plastic mesh and variation in frequency of application of 20% sucrose) and exposure to fluoride dentifrice. Themineral content and lesion depth were evaluated by transverse microradiography (TMR). 1) The TiF4 varnishes (1.95 and 2.45% F) released more F- compared to NaF varnishes in both water and artificial saliva, and significant difference was found in the first 6h (p<0.0001). 2) The TiF4 caused change in the HAP structure and induced the formation of new compounds such as TiO2 and Ti (HPO4)2. 3) The TiF4 varnishes induced the formation of a coating layer rich in Ti and F, with microcracks in its extension, on the enamel...

Efeito do cranberry e seus componentes sobre a dentina submetida à erosão / Effect of cranberry on dentin erosion challenge

Existem indícios de que o Cranberry e a Proantocianidina (polifenol desta fruta) tenham potencial para inibir as metaloproteinases da matriz (MMPs) que degradam o colágeno dentinário, porém estes agentes não foram adequadamente avaliados em estudos sobre erosão dentária. Assim, o objetivo desta tese será avaliar o papel do extrato de Cranberry e do seu principal agente ativo isolado (proantocianidina) aplicados na forma de gel tópico na inibição da degradação da matriz orgânica da dentina desmineralizada e consequentemente na minimização do desgaste da dentina submetida à erosão. Para isso, foram realizados 3 trabalhos buscando avaliar os seguintes objetivos específicos: Artigo 1- Verificar se diferentes concentrações de géis a base de extrato de Cranberry e de Proantocianidina apresentam papel inibitório do desgaste de espécimes dentinários submetidos à erosão in vitro, tendo como controles um grupo placebo e a clorexidina; Artigo 2- Avaliar o possível efeito protetor in situ de um gel de Cranberry aplicado à dentina submetida a ciclagem erosiva, quando comparados a grupos placebo e clorexidina; Artigo 3- Avaliar o efeito de diferentes concentrações de géis de Proantocianidina, aplicados por diferentes tempos de aplicação na minimização do desgaste da dentina exposta a desafios erosivos in vitro. Os resultados desta tese mostraram que os géis experimentais testados tiveram efeitos superiores aos grupos placebos e semelhantes ao controle positivo (clorexidina) na minimização do desgaste da dentina submetida à erosão. Além disso, nas concentrações avaliadas, não foi possível encontrar uma relação dose e tempo de aplicação/ resposta na prevenção da erosão. Assim, considerando os resultados apresentados na presente tese pôde- se concluir de forma geral que os géis de Cranberry e Proantocianidina foram efetivos na diminuição do desgaste dentário quando a dentina é exposta a desafio erosivo.(AU)

There are indications that the Cranberry and Proanthocyanidin (this fruit's polyphenol) have the potential to inhibit matrix metalloproteinases (MMPs) that degrade collagen from dentin, however these agents have not been adequately evaluated in studies of dental erosion. Thus, the aim of this thesis was to evaluate the role of Cranberry extract and its isolated main active agent (proanthocyanidin) applied as topical gel in the inhibition of organic matrix degradation and thus minimizing wear of dentin subjected to erosion. For that, three studies were conducted to assess the following specific objectives: Paper 1- Evaluate different gel concentrations of Cranberry extract and Proanthocyanidin on diminishing dentin erosion progression in vitro, having as control a placebo group and chlorhexidine; Paper 2- Evaluate the possible protective in situ effect of a Cranberry gel applied to dentin subjected to erosive cycling, when compared to placebo groups and chlorhexidine, Paper 3- Evaluate the effect of different concentrations of proanthocyanidin gels, applied by different application times in minimizing the wear of dentin exposed to erosive challenge in vitro. The results of this thesis showed that the tested experimental gels had greater effect than placebos groups and similar to the positive control (chlorhexidine) in minimizing the wear of dentin subjected to erosion. Furthermore, in the concentrations evaluated, it was not possible to find a dose and time of application / response in the prevention of erosion. Thus, considering the results presented in this thesis, it could be concluded that in general the gels of Cranberry and Proanthocyanidin were effective in decreasing the tooth wear when the dentin was exposed to erosive challenge.(AU)

Avaliação da interação do TiF4 com a hidroxiapatita e esmalte bem como o efeito sobre a des-remineralização do esmalte dentário com lesão de cárie artificial in situ / Interaction of TiF4 varnish with enamel and hydroxyapatite and its effect on the de-remineralization of pre-demineralized enamel in situ

Este estudo teve como o objetivo avaliar in vitro o efeito do tratamento com vernizes de tetrafluoreto de titânio (TiF4) e fluoreto de sódio (NaF) (ambos com 0,95%, 1,95% e 2,45% F) sobre 1) a quantidade de F- liberada em água deionizada e saliva artificial, por período de 12h; 2) os tipos de compostos formados pela interação com a hidroxiapatita (neste caso, com soluções ao invés dos vernizes); 3) a porcentagem de elementos presentes na superfície do esmalte bovino e humano, hígidos (H) e desmineralizados (DES); 4) a quantidade de CaF2 sobre a superfície do esmalte bovino e humano, H e DES. Adicionalmente, 5) o efeito dos vernizes de TiF4 e NaF sobre a remineralização do esmalte bovino, em diferentes condições in situ, foi avaliado. Para etapa 1, a liberação de F- foi quantificada por eletrodo de íon específico. Na etapa 2, pó de HAP produzido por precipitação foi tratado com soluções fluoretadas. Os compostos formados foram avaliados por espectroscopia de infravermelho e difração de raios-X. Nas etapas 3 e 4, as superfícies tratadas com os vernizes foram analisadas por MEV- EDAX e por biópsias básicas, respectivamente. Vinte voluntários (n final=17) participaram do ensaio in situ com 3 fases cruzadas, os quais utilizaram aparelhos palatinos contendo amostras de esmalte bovino desmineralizadas tratadas com vernizes de TiF4, NaF ou placebo. As amostras foram submetidas a diferentes condições de des-remineralização (presença ou não de tela plástica; variação da frequência de aplicação de sacarose 20%) e à exposição ao dentifrício fluoretado. O volume mineral e profundidade da lesão foram avaliados por microradiografia transversal (TMR). 1) Os vernizes de TiF4 (1,95 e 2,45% F) liberaram mais fluoreto comparados aos vernizes de NaF tanto em água como em saliva artificial, sendo a diferença significativa nas primeiras 6h de contato (p<0,0001). 2) O TiF4 causou alteração na estrutura da HAP e induziu à formação de novos compostos como TiO2 e Ti (HPO4)2. 3)...

This study aimed to evaluate the in vitro effect of titanium tetrafluoride (TiF4) and sodium fluoride (NaF) varnishes (both with 0.95%, 1.95%, 2.45% F) treatment on the 1) F- release in deionized water and artificial saliva for 12h period; 2) types of compounds formed by the interaction with hydroxyapatite (in this case, solutions were tested instead of varnishes); 3) percentage of the elements on bovine and human, sound (S) and demineralized (DE), enamel surface; 4) F- uptake (CaF2 deposition) on human and bovine, S and DE, enamel surface. Additionally, 5) the effect of TiF4 and NaF varnishes on bovine enamel remineralization, in different in situ conditions, was assessed. In study 1, the F- release was measured by ion specific electrode. In study 2, HAP powder, produced by precipitation, was treated with fluoride solutions. The compounds formed were evaluated by infrared spectroscopy and X-ray diffraction. In studies 3 and 4, the surfaces treated with the varnishes were analyzed by SEM-EDAX and basic biopsies, respectively. Twenty volunteers (final n=17) participated of the in situ study with 3 crossover phases, in which they wore palatal appliances containing bovine demineralized enamel samples treated with TiF4, NaF or placebo varnishes. The samples were subjected to different de-remineralization conditions (presence or absence of plastic mesh and variation in frequency of application of 20% sucrose) and exposure to fluoride dentifrice. Themineral content and lesion depth were evaluated by transverse microradiography (TMR). 1) The TiF4 varnishes (1.95 and 2.45% F) released more F- compared to NaF varnishes in both water and artificial saliva, and significant difference was found in the first 6h (p<0.0001). 2) The TiF4 caused change in the HAP structure and induced the formation of new compounds such as TiO2 and Ti (HPO4)2. 3) The TiF4 varnishes induced the formation of a coating layer rich in Ti and F, with microcracks in its extension, on the enamel...

Fluoride stability in dentifrices stored in schools in a town of northern Brazil

The aim of this study was to analyze fluoride stability in dentifrices stored during nine months in schools from Careiro da Várzea, State of Amazonas, Brazil. Analysis of total fluoride concentration, total soluble fluoride, and ionic fluoride in the dentifrice samples was performed in four different time periods: at the time of purchase (baseline); after three months, after six months, and after nine months of storage. Fluoride concentration was determined using a specific electrode (Orion 96-09) connected to an ion analyzer (Orion A-720) and calibrated with fluoride standard solutions containing 2.0 to 32.0 ppm F. The results obtained during the measurements were analyzed by analysis of variance (one-way ANOVA), followed by Tukey’s test for comparison of the means. After nine months of storage, total soluble fluoride, the active form of fluoride, decreased by 21.9%. As total soluble fluoride was below the minimum required for anticaries efficacy (1,000 ppm F) in the fourth analysis, it may be concluded that anticaries potential decreased with storage time.

Fluoride varnishes with calcium glycerophosphate: fluoride release and effect on in vitro enamel demineralization

The aims of this study were (1) to assess the amount of fluoride (F) released from varnishes containing calcium glycerophosphate (CaGP) and (2) to assess the effect of the experimental varnishes on in vitrodemineralization. Six test groups using 5 varnishes: base varnish (no active ingredients); Duraphat® (2.26% NaF); Duofluorid® (5.63% NaF/CaF2); experimental varnish 1 (1% CaGP/5.63% NaF/CaF2); experimental varnish 2 (5% CaGP/5.63% NaF/CaF2); and no varnish were set up. In stage 1, 60 acrylic blocks were randomly distributed into 6 groups (n = 10). Then 300 µg of each varnish was applied to each block. The blocks were immersed in deionized water, which was changed after 1, 8, 12, 24, 48 and 72 hours. Fluoride concentration in the water was analyzed using a fluoride electrode. In stage 2, 60 bovine enamel samples were distributed into 6 groups (n = 10), and treated with 300 µg of the respective varnish. After 6 h the varnish was removed and the samples were subjected to a 7-day in vitro pH cycle (6 h demineralization/18 h remineralization per day). The demineralization was measured using surface hardness. The results showed that both experimental varnishes released more fluoride than Duofluorid® and Duraphat® (p < 0.05), but Duraphat® showed the best preventive effect by decreasing enamel hardness loss (p < 0.05). Therefore, we conclude that even though (1) the experimental varnishes containing CaGP released greater amounts of F, (2) they did not increase in the preventive effect against enamel demineralization.