Enamel Caries Lesion Depth Obtained by Optical Coherence Tomography and Transverse Microradiography: A Comparative Study.

INTRODUCTION: Visual imaging of subsurface caries lesions is of vital interest in dentistry, which can be obtained by invasive radiography technique as well as by available non-destructive imaging approaches. Thus, as a first step toward the development of a new innovative approach, Spectral-domain optical coherence tomography (SD-OCT) was applied to detect the lesion depth in comparison to the established reference technique (transverse microradiography [TMR]). METHODS: Bovine enamel specimens were demineralized for 5 days, following previous studies. For OCT, the resulting artificial lesions were scanned three-dimensionally (SD-OCT) and semi-automated measured (CarLQuant). For TMR, specimens were sectioned and the lesion depth was manually determined (Inspektor Research System). RESULTS: The range of lesion depth detected with OCT was 24.0-174.0 µm (mouth rinse study), 18.0-178.0 µm (toothpastes study) and with TMR 59.2-198.0 µm (mouth rinse study), 33.2-133.4 µm (toothpastes study). We found a strong correlation between both methods in terms of lesion depth (Spearman rankwith outlierp < 0.001, Rho = 0.75, Spearman rankwithout outlierp = 0.001, Rho = 0.79). The two methods produce similar results (Passing-Bablok regression, 1.16). As deeper is the lesion, the smallest is the difference between both methods as indicated by Bland-Altman-plots. CONCLUSION: Especially in the case of deep lesions, the values obtained by both methods are in agreement, and OCT can potentially substitute TMR to detect and assess lesion depth with the benefit of being non-destructive.

Organic volume and permeability variations in the surface layer of artificial and natural enamel carious lesions.

OBJECTIVES: Test the hypothesis the type of enamel caries (natural, artificial induced by gel, and artificial induced by acid solutions) affect the organic volume and the permeability of the surface layer in enamel caries lesions. DESIGN: Artificial enamel caries, induced by either acidic solution (organic-poor; Group 1) or acidic gel (organic-rich; Group 2), and natural non-cavitated inactive approximal enamel caries lesions (NEC; Group 3) were obtained, from which longitudinal ground sections were prepared. Measurements of the mineral (Vmin) (by microradiography), and water (α) and organic (ß) volumes (by optical birefringence) were obtained at three points in the surface layer (n = 30/group). RESULTS: The main outcomes were the ratio between experimental ß by predicted ß (ß Ratio) and the ratio between experimental and predicted permeabilities (αd Ratio). ß Ratio in Group 1 was lower than in Groups 2 (Cohen's d: -1.81; 95% CI:-1.45,-2.32; p < 0.001) and 3 (Cohen's d: -0.71; 95% CI:-0.27,-1.18; p = 0.004), and Group 2 surpassed Group 3 (Cohen's d: 0.49; 95% CI:0.07,0.94; p = 0.03). αd Ratio in Group 1 was higher than in Groups 2 (Cohen's d: 1.86; 95% CI:1.49,2.33; p < 0.001) and 3 (Cohen's d: 0.60; 95% CI:0.18,1.14; p = 0.01), and Group 3 surpassed Group 2 (Cohen's d: 0.61; 95% CI:0.23,1.07; p = 0.01). CONCLUSIONS: The highest organic volume and the lowest permeability occurred at the surface layer of gel-induced artificial enamel caries lesions, which should be preferred in in vitro studies on de- and remineralization and resin infiltration.

Effects of silver diamine fluoride on demineralization protection after a secondary acid challenge

Abstract Objective This investigation describes the effects of 5% sodium fluoride varnish and 38% silver diamine fluoride on demineralization protection of human enamel lesions of three different severities after a secondary acid challenge. Study design Specimens underwent color and enamel surface microhardness change measurements after demineralization and treatment events. Transverse microradiography was conducted following the secondary demineralization. Results After treatments, enamel surface microhardness change showed that 24-hour lesions treated with fluoride varnish had less rehardening than 24-hour lesions treated with silver diamine fluoride (p<0.05), whereas 144-hour lesions from both treatment groups showed a beneficial decrease in surface microhardness change that was markedly better in samples treated with silver diamine fluoride (p<0.05). After the secondary demineralization, 24- and 144-hour lesions treated with silver diamine fluoride showed a sustained beneficial decrease in enamel surface microhardness change when compared to fluoride varnish-treated samples of the corresponding lesion severity (p<0.05). Transverse microradiography showed no difference between fluoride varnish- and silver diamine fluoride-treated samples of any corresponding lesion severity, indicating that remineralization in both fluoride varnish- and silver diamine fluoride-treated samples was proportional to each other after a secondary acid challenge. Conclusions Using silver diamine fluoride may have comparable benefits to fluoride varnish in mineral loss prevention.

Monitoring enamel caries on resin-treated occlusal surfaces using quantitative light-induced fluorescence: an in vitro study.

The aim of this study is to evaluate the ability of quantitative light-induced fluorescence (QLF) to monitor enamel caries lesions of different severity stages located on the occlusal surfaces of permanent teeth before and after treatment with resin infiltrant. Sixty extracted permanent teeth had one occlusal site selected and were categorized according to the International Caries Detection and Assessment System (ICDAS) criteria. The teeth were divided into three groups (n = 20): ICDAS 1, ICDAS 2, and ICDAS 3. The teeth were assessed by a trained examiner using QLF in two phases: (A) before and (B) after treatment with resin infiltrant. The caries lesions were evaluated using the following QLF parameters: area (mm2); ΔF, fluorescence loss (%); and ΔQ, fluorescence loss integrated over the lesion area (%*mm2). The resin infiltrant (Icon™) was applied on the occlusal surface following the manufacturer's recommendations. The teeth were then sectioned and prepared for polarized light microscopy analysis. The penetration of resin infiltrant was measured with ImageJ. The groups showed a statistically significant difference in all QLF parameters before and after caries infiltration, with the reduction of fluorescence values posttreatment (p < 0.05). Infiltrant penetration was observed in all groups, with a statistical difference between all groups (p < 0.05). The reduction in QLF parameters after resin infiltration suggests that QLF is able to monitor enamel caries lesions of different severity stages located on the occlusal surfaces of permanent teeth before and after treatment with resin infiltrant.

High Amount of Organic Matter during Caries Formation Reduces Remineralization and Resin Infiltration of Enamel Caries.

The amount of organic material in the cariogenic environment correlates with the amount of organic material incorporated in carious enamel. The incorporated organic material may be expected to reduce the pore volumes available for remineralization and resin infiltration, but these expected outcomes have not yet been quantified. We tested the effect of the amount of organic content in the cariogenic agent on remineralization and the resin-occluded pore volume in artificial subsurface enamel caries. An acid gel (organic-rich; G1) and an aqueous solution (organic-poor; G2) were used to induce subsurface lesions in human enamel. Undemineralized histological sections were prepared, microradiographed, and then submitted to resin infiltration in vitro. The enamel component volumes (mineral, organic, remineralizable [total water volume], loosely and firmly bound water volumes, and resin-occluded volume) were measured (by microradiography and polarizing microscopy) at histological sites (n = 38, G1; n = 34, G2). The main outcomes were the differences between the experimental and the predicted volumes (Δremineralizable and Δresin-occluded volumes). Resin infiltration was confirmed by confocal scanning laser microscopy. Compared to G2, G1 presented more incorporated organic volume and lower Δremineralizable volume (p = 0.003; Hedges g = 0.66; power = 0.87), a lower increase in loosely bound water volume (p = 0.0013; Hedges g = 0.74; power = 0.93), a lower remineralization volume in the surface layer (p = 0.017; Hedges g = 0.68; power = 0.8), and a lower Δresin-occluded volume (p = 0.0015; Hedges g = 0.73; power = 0.92). In conclusion, the higher amount of organic matter in the cariogenic gel negatively affected remineralization and the resin-occluded volume in subsurface lesions.

Effect of Myracrodruon urundeuva and Qualea grandiflora extracts on viability and activity of microcosm biofilm and prevention of enamel demineralization in vitro / Efeito de extratos de Myracrodruon urundeuva All. e Qualea grandiflora Mart. sobre a viabilidade e atividade de biofilme microcosmo e na prevenção da desmineralização do esmalte in vitro

The objective of this study was to evaluate the antimicrobial and anti-caries effects of two plant extracts. The first chapter dealt with a review of the literature whose objective was to discuss the antimicrobial potential of Brazilian natural agents on the biofilm related to dental caries and gingivitis/periodontal disease. The research of the articles was carried out using PubMed. We found a total of 23 papers. Most of the studies were performed using planktonic microorganisms or under clinical trials. Nineteen articles were focused on cariogenic bacteria. From these nineteen articles, eleven were also about periodontopathogenic bacteria. Four studies addressed only periodontopathogenic bacteria. The most tested Brazilian natural agents were green propolis, essential oils of Lippia sidoides and Copaifera sp. Most of the tested agents showed similar results when compared to positive control (essential oils and extracts) or better effect than negative control (green propolis). More studies involving protocols closer to the clinical condition and the use of response variables that allows understanding the mechanism of action of natural agents are necessary before the incorporation of these natural agents into dental products. The second chapter aimed to test the effect of the hydroalcoholic extracts of Myracrodruon urundeuva All. and Qualea grandiflora Mart. leaves on the viability of the microcosm biofilm and on the prevention of enamel demineralization. The microcosm biofilm was produced on bovine enamel, using human saliva pool mixed with McBain saliva (0.2% sucrose) for 14 days. The biofilm was treated daily with the extracts for 1 min. M. urundeuva at 100, 10 and 0.1 µg/ml and Q. grandiflora at 100 and 0.1 µg/ml reduced cell viability similarly to the positive control and significantly more than negative control. M. urundeuva at 1000, 100 and 0.1 µg/ml were able to reduce the counting formation unit-CFU counting of lactobacilli sp. and Streptococcus mutans, while Q. grandiflora at 1000 and 1.0 µg/ml significantly reduced the S. mutans CFU counting. On the other hand, the natural extracts did not reduce the production of extracellular polyssacharides, lactic acid and the development of enamel caries lesions. The third chapter aimed to evaluate the effect of hydroalcoholic extracts of M. urundeuva and Q. grandiflora (alone or combined) on the viability of S. mutans biofilm and the prevention of enamel demineralization. S. mutans strain (ATCC 21175) was reactivated in BHI broth. Minimum inhibitory concentration, minimum bactericidal concentration, minimum biofilm inhibitory concentration and minimum biofilm eradication concentration were determined to choose the concentrations to be tested under the biofilm model. S. mutans biofilm (5x105 CFU/ml) was produced on bovine enamel using McBain saliva with 0.2% sucrose for 3 days. The biofilm was treated daily with the extracts for 1 min. M. urundeuva (isolated or combined) at concentrations equal or higher than 0.625 mg/ml was able to reduce the bacteria viability, whereas Q. grandiflora extract alone showed antimicrobial effect at 5 mg/ml only (p<0.05). On the other hand, none of the extracts was able to reduce the development of enamel caries lesions. Despite the tested natural extracts have antimicrobial effect; they are unable to prevent caries in enamel.(AU)

O objetivo foi avaliar os efeitos antimicrobiano e anti-cárie de dois extratos de plantas. O primeiro capítulo se referiu a uma revisão da literatura cujo objetivo foi discutir o potencial antimicrobiano dos agentes naturais brasileiros sobre o biofilme relacionado à cárie dentária e à gengivite/doença periodontal. A pesquisa dos artigos foi realizada usando o PubMed. Foram encontrados 23 trabalhos. A maioria dos estudos foi realizada utilizando microorganismos na fase planctônica ou ensaios clínicos. Dezenove artigos foram focados em bactérias cariogênicas. Dos dezenove artigos, onze também eram sobre bactérias periodontopatogênicas. Quatro estudos abordaram apenas bactérias periodontopatogênicas. Os agentes naturais brasileiros mais testados foram própolis verde, óleos essenciais de Lippia sidoides e Copaifera sp. Os agentes testados apresentaram resultados similares quando comparados ao controle positivo (óleos essenciais e extratos) ou melhor efeito que o controle negativo (própolis verde). Mais estudos próximos da condição clínica e o uso de variáveis de resposta que permitam entender o mecanismo de ação são necessários, para permitir a incorporação desses agentes naturais em produtos odontológicos. O segundo capítulo teve como objetivo testar o efeito dos extratos hidroalcoólicos de Myracrodruon urundeuva All. e Qualea grandiflora Mart. sobre a viabilidade do biofilme microcosmo e na prevenção da desmineralização do esmalte. O biofilme microcosmo foi produzido em esmalte bovino, utilizando pool de saliva humana misturada à saliva de McBain (0,2% de sacarose) durante 14 dias. O biofilme foi tratado diariamente com os extratos durante 1 min. M. urundeuva a 100, 10 e 0,1 µg/ml e Q. grandiflora a 100 e 0,1 µg/ml reduziram a viabilidade dos microrganismos de forma semelhante ao controle positivo e significativamente maior do que o controle negativo. M. urundeuva a 1000, 100 e 0,1 µg/ml foi capaz de reduzir a contagem de Unidade formadora de colônia-UFC para Lactobacilos totais e Streptococcus mutans, enquanto a Q. grandiflora a 1000 e 1,0 µg/ml reduziu significativamente a contagem de UFC para S. mutans. Os extratos naturais não conseguiram reduzir a produção de polissacarídeos extracelulares-PEC, ácido lático e o desenvolvimento da lesão cariosa em esmalte. O terceiro capítulo teve como objetivo avaliar o efeito dos extratos hidroalcoólicos de M. urundeuva. e Q. grandiflora (sozinhos ou combinados) sobre a viabilidade do biofilme de S. mutans e na prevenção da desmineralização do esmalte. Cepa de S. mutans (ATCC 21175) foi reativada em caldo BHI. Concentração inibitória mínima, concentração bactericida mínima, concentração inibitória mínima de biofilme e concentração de erradicação mínima de biofilme foram determinadas para escolher as concentrações a serem testadas sob o modelo de biofilme. O biofilme de S. mutans (5x105 CFU/ml) foi produzido em esmalte bovino, utilizando saliva de McBain com 0,2% de sacarose durante 3 dias. O biofilme foi tratado diariamente com os extratos durante 1 min. M. urundeuva (isolada ou combinada) nas concentrações iguais ou superiores a 0,625 mg/ml foi capaz de reduzir a viabilidade das bactérias, enquanto que o extrato da Q. grandflora apresentou efeito antimicrobiano somente a 5 mg/ml (p<0,05). Nenhum dos extratos reduziu o desenvolvimento da lesão da cárie. Apesar dos extratos naturais terem efeito antimicrobiano, são incapazes de prevenir o desenvolvimento da lesão cariosa em esmalte.(AU)

Analysis of the antimicrobial and anti-caries effects of TiF4 varnish under microcosm biofilm formed on enamel

Abstract Titanium tetrafluoride (TiF4) is known for interacting with enamel reducing demineralization. However, no information is available about its potential antimicrobial effect. Objectives This study evaluated the antimicrobial and anti-caries potential of TiF4 varnish compared to NaF varnish, chlorhexidine gel (positive control), placebo varnish and untreated (negative controls) using a dental microcosm biofilm model. Material and Methods A microcosm biofilm was produced on bovine enamel previously treated with the varnishes, using inoculum from human saliva mixed with McBain saliva, under 0.2% sucrose exposure, for 14 days. All experiments were performed in biological triplicate (n=4/group in each experiment). Factors evaluated were: bacterial viability (% dead and live bacteria); CFU counting (log10 CFU/mL); and enamel demineralization (transverse microradiography - TMR). Data were analysed using ANOVA/Tukey's test or Kruskal-Wallis/Dunn's test (p<0.05). Results Only chlorhexidine significantly increased the number of dead bacteria (68.8±13.1% dead bacteria) compared to untreated control (48.9±16.1% dead bacteria). No treatment reduced the CFU counting (total microorganism and total streptococci) compared to the negative controls. Only TiF4 was able to reduce enamel demineralization (ΔZ 1110.7±803.2 vol% μm) compared to both negative controls (untreated: ΔZ 4455.3±1176.4 vol% μm). Conclusions TiF4 varnish has no relevant antimicrobial effect. Nevertheless, TiF4 varnish was effective in reducing enamel demineralization under this model.

Evaluation of the effect of different commercial mouthrinses on the viability and activity of microcosm biofilm and on enamel demineralization / Avaliação do efeito de diferentes enxaguatórios bucais comerciais sobre a viabilidade e atividade de biofilme microcosmo e a desmineralização do esmalte

The aim of this study was to evaluate the antimicrobial and anticaries effects of different commercial mouthrinses. The first chapter is about a review of literature whose the aim was to discuss the antimicrobial potential of different mouthrinses in respect to the control of dental caries and periodontal disease. The search of papers was conducted using PubMed and the keywords: "antimicrobial agent" or "antiplaque agent," "dental biofilm" and "dental caries" or "periodontal disease" or "gingivitis". We found a total of 22 papers (2011-2015). The main active agents tested were: CHXChlorhexidine, CPC-cetylpyridinium chloride and EO-Essential oils (alcohol/or alcohol-free). CHX was compared to EO in 6 studies, showing superiority in 3 studies, similarity in 1 study and inferiority in 2 studies. CPC has shown lower effect in plaque reduction compared to CHX and EO. More clinical studies are needed for better understanding the mechanism of action and the differences in performance among the antiplaque agents. The second chapter has as aim to compare the antimicrobial and anticaries effects of six commercial mouthrinses (PerioGard®, Noplak® Max, Oral-B® Complete, Listerine® Zero, Malvatricin® Plus and Cepacol® Plus Advanced) under a microcosm biofilm model formed on enamel. A microcosm biofilm was produced on bovine enamel, using inoculum from pooled human saliva mixture with McBain saliva (with 0.2% sucrose), for 14 days. The biofilm was treated with the mouthrinses daily (1 min). The bacterial viability (% death), lactic acid production (mmol/l), the colony-forming unit (CFU) counting for total microorganisms, lactobacilli, total streptococci and mutans streptococci (log10 CFU/mL) and the extracellular polysaccharides production (EPS, mg/g) were quantified in the biofilm. The degree of enamel demineralization was analyzed using transverse microradiography-TMR (%min vol. µm). Oral-B® Complete, Listerine® Zero and Malvatricin® Plus had the greatest effect on the reduction of biofilm viability (69-75% dead cells vs. 13% in the control, p<0.0001). On the other hand, the lactic acid production was significantly reduced by PerioGard®, Noplak® Max and Listerine® Zero compared to control (69% reduction, p<0.0001). There were no significant differences among the mouthrinses in respect to the CFU counting and EPS production. The enamel demineralization was significantly reduced by PerioGard®, Noplak® Max and Malvatricin® Plus compared to control (74% reduction, p<0.0001). Therefore, the commercial mouthrinses have different antimicrobial and anticaries effects. The mouthrinses containing chlorexidine or Malva sylvestris (with F, triclosan and xylitol) had the best anticaries effect under this model.(AU)

O objetivo deste estudo foi avaliar os efeitos antimicrobiano e anticárie de diferentes enxaguatórios bucais comerciais. O primeiro capítulo se refere a uma revisão da literatura cujo objetivo foi discutir o potencial antimicrobiano de diferentes enxaguatórios em relação ao controle de cárie dentária e doença periodontal. A pesquisa dos artigos foi realizada usando o PubMed e as palavras-chave: "agente antimicrobiano" ou "agente antiplaca", "biofilme dental" e "cárie dentária" ou "doença periodontal" ou "gengivite". Encontramos um total de 22 artigos (2011-2015). Os principais agentes ativos testados foram: Clorexidina-CHX, cloreto de cetilpiridínio- CPC e óleos essenciais OE (álcool / ou sem álcool). A CHX foi comparada ao OE em 6 estudos, mostrando superioridade em 3 estudos, similaridade em 1 estudo e inferioridade em 2 estudos. CPC mostrou menor efeito na redução da placa em comparação com CHX e OE. Mais estudos clínicos são necessários para uma melhor compreensão do mecanismo de ação e as diferenças no desempenho entre os agentes antiplaca. O segundo capítulo teve como objetivo avaliar os efeitos antimicrobiano e anticárie de seis enxaguatórios comerciais (PerioGard®, Noplak® Max, Oral-B® Complete, Listerine® Zero, Malvatricin® Plus e Cepacol® Plus Advanced) sobre um modelo de biofilme de microcosmo formado no esmalte. O biofilme de microcosmo foi produzido em esmalte bovino, utilizando o inóculo da mistura de saliva humana com saliva de McBain (com 0,2% de sacarose), durante 14 dias. O biofilme foi tratado com enxaguatórios diariamente (1 min). A viabilidade bacteriana (% de morte), a produção de ácido láctico (mmol/l), as unidades formadoras de colônias (UFC) foram contadas para microrganismos totais, lactobacilos, estreptococos totais e Streptococcus mutans (log10 UFC/mL) e a produção de polissacarídeos extracelulares (PEC, mg/g) foram quantificados no biofilme. O grau de desmineralização do esmalte foi analisado utilizando a microrradiografia transversal-TMR (%min vol. µm). Oral-B® Complete, Listerine® Zero e Malvatricin® Plus tiveram o maior efeito na redução da viabilidade do biofilme (69-75% de morte celular vs 13% no controle, p<0,0001). Por outro lado, a produção de ácido lático foi significativamente reduzida por PerioGard®, Noplak® Max e Listerine® Zero comparado ao controle (redução de 69%, p<0,0001). Não houve diferenças significativas entre os enxaguatórios em relação à contagem de UFC e à produção de PEC. A desmineralização do esmalte foi significativamente reduzida pelo PerioGard®, Noplak® Max e Malvatricin® Plus em comparação ao controle (redução de 74%, p<0,0001). Portanto, os enxaguatórios bucais comerciais têm diferentes efeitos antimicrobiano e anticárie. Os enxaguatórios contendo clorexidina ou Malva sylvestris (com F, triclosan e xilitol) tiveram o melhor efeito anticárie neste modelo.(AU)

Evaluation of polymerization characteristics and penetration into enamel caries lesions of experimental infiltrants.

OBJECTIVES: To evaluate the properties of experimental infiltrant blends by comparing them with the commercial infiltrant Icon(®) and penetration homogeneity into enamel caries lesions. METHODS: Groups were set up as follows: G1 (TEGDMA 100%); G2 (TEGDMA 80%, Ethanol 20%); G3 (TEGDMA 80%, HEMA 20%); G4 (TEGDMA 75%, BisEMA 25%); G5 (TEGDMA 60%, BisEMA 20%, Ethanol 20%); G6 (TEGDMA 60%, BisEMA 20%, HEMA 20%); G7 (TEGDMA 75%, UDMA 25%); G8 (TEGDMA 60%, UDMA 20%, Ethanol 20%); G9 (TEGDMA 60%, UDMA 20%, HEMA 20%) and Icon(®). Ten specimens were comprised by each group for the following tests (n=10): degree of conversion (DC), elastic modulus (EM), Knoop hardness (KH), and softening ratio (SR). Infiltrant penetration was evaluated using confocal microscopy (CLSM). Data were subjected to two-way ANOVA and a Tukey's test (5%). Data comparing experimental materials and Icon(®) were analysed using ANOVA and Dunnett's test (5%). RESULTS: The highest DC values were found in G1, G7, G8, and G9. The lowest DC values were found in G2, G4, G5, and G6. EM and KHN were significantly lower in HEMA and with ethanol addition for all blends, except for G9. There was no significant difference among the groups regarding SR, and it was not possible to take KHN readings of G2, G5, and G8 after storage. There was no significant difference among groups for infiltrant penetration into enamel lesions. CONCLUSIONS: The addition of hydrophobic monomers and solvents into TEGDMA blends affected DC, EM, and KHN. UDMA added to TEGDMA resulted in an increase in DC, EM, and KHN. Overall, solvents added to monomer blends resulted in decreased properties. The addition of hydrophobic monomers and solvents into TEGDMA blends does not improve the penetration depth of the infiltrants.