Abstract Dental hard tissue conditions can be of pre- or post-eruptive nature, such as enamel fluorosis and erosive tooth wear (ETW), respectively. Dental enamel fluorosis is caused by the chronic and excessive intake of fluoride during enamel development, leading to increased fluoride concentration and increased porosity. ETW has become a common clinical condition and often impairs dental function and aesthetics. This in vitro study tested the hypothesis that fluorotic enamel presents different susceptibility to dental erosion-abrasion. It consisted of a 3×3×2 factorial design, considering a) fluorosis severity: sound (TF0), mild (TF1-2), moderate (TF3-4); b) abrasive challenge: low, medium, and high; and c) erosive challenge: yes or no. A total of 144 human teeth were selected according to the three fluorosis severity levels (n=48), and subdivided into six groups (n = 8) generated by the association of the different erosive and abrasive challenges. Enamel blocks (4×4 mm) were prepared from each tooth and their natural enamel surfaces subjected to an erosion-abrasion cycling model. After cycling, the depth of the lesions in enamel was assessed by profilometry. ANOVA showed that the three-way and two-way interactions among the factors were not significant (p > 0.20). Enamel fluorosis level (p=0.638) and abrasion level (p = 0.390) had no significant effect on lesion depth. Acid exposure caused significantly more enamel surface loss than water (p < 0.001). Considering the limitations of this in vitro study, fluorosis did not affect the susceptibility of enamel to dental erosion-abrasion.

Abstract The purpose of this in vitro study was to test the hypothesis that fluoride treatment can prevent dental erosion on fluorotic enamel of different severities. It followed a 3×2 factorial design, considering a) fluorosis severity: sound (TF0, Thylstrup-Fejerskov Index), mild (TF1-2), moderate (TF3-4); and b) fluoride treatment: 0 (negative control) and 1150ppmF. Human molars with the three fluorosis severities (n=16, each) were selected and randomly assigned to the two fluoride treatments (n=8). Enamel blocks (4×4mm) were prepared from each tooth and subjected to a dental erosion cycling model, for 10 days. The daily cycling protocol consisted of erosive challenges (1% citric acid, pH 2.4), interspersed by periods of immersion in artificial saliva, and three 2-minute treatments with either 0 or 1150ppm F. The enamel volume loss (mm3) was calculated by subtracting values obtained by microtomography before and after cycling. Two-Way ANOVA showed no significant interaction between fluorosis severity and fluoride treatment (p=0.691), and no significant effect for either fluorosis severity (TF0 mean±standard-deviation: 13.5(10-2±0.42(10-2, TF1-2: 1.50(10-2±0.52(10-2, TF3-4: 1.24(10-2±0.52(10-2, p=0.416) or treatment (0ppmF: 1.49(10-2±0.53(10-2; 1150ppmF: 1.21(10-2±0.42(10-2; p=0.093), when evaluated independently. Considering the limitations of this in vitro study, the presence and severity of fluorosis in enamel do not appear to affect its susceptibility to dental erosion. Fluoride treatment was not effective in preventing the development of dental erosion in both sound and fluorotic enamel substrates under our experimental conditions.

Resumo Este estudo in vitro foi testou a hipótese de que o tratamento com flúor pode prevenir a erosão dentária no esmalte fluorótico de diferentes severidades. O objetivo deste estudo foi: investigar o efeito protetor dos fluoretos contra a erosão e abrasão simuladas no esmalte fluorótico. Seguiu um desenho fatorial 3×2, considerando a) severidade da fluorose em 3 níveis: hígido (TF0, Índice Thylstrup-Fejerskov), suave (TF1-2), moderada (TF3-4); b) tratamento com flúor: 0 (controle negativo) e 1150ppmF. Molares humanos com as três severidades de fluorose (n=16, cada) foram selecionados e distribuídos aleatoriamente para os dois tratamentos com flúor (n=8). Blocos de esmalte (4×4mm) foram preparados a partir de cada dente e submetidos a um modelo de ciclo de erosão dentária, por 10 dias. O protocolo de ciclagem diária consistiu em seis desafios erosivos de 5 minutos (1% de ácido cítrico, pH 2,4), intercalados por seis períodos de imersão em saliva artificial e três tratamentos de 2 minutos com 0 ou 1150ppmF. O volume do esmalte perdido foi calculado subtraindo o perfil superficial 3D obtido por microtomografia antes e depois da ciclagem. A ANOVA de dois fatores não mostrou interação significativa entre a severidade da fluorose e o tratamento com flúor (p = 0,691) e nenhum efeito significativo para a severidade da fluorose (TF0 média+/desvio padrão: 13,5(10-2±0,42(10-2, TF1-2: 1,50(10-2±0,52(10-2, TF3-4: 1,24(10-2±0,52(10-2, p=0,416) ou tratamento (0: 1,49(10-2±0,53(10-2; 1150ppmF: 1,21(10-2±0,42(10-2, p=0,093), quando avaliados independentemente. Considerando as limitações deste estudo in vitro, a presença e severidade da fluorose no esmalte não parece afetar sua suscetibilidade à erosão dentária. O tratamento com flúor não foi eficaz na prevenção do desenvolvimento da erosão dentária em esmalte hígido e fluorótico, sob as condições experimentais utilizadas.

Abstract This study evaluated the orofacial antinociceptive effect of (S)-(-)-perillyl alcohol (PA) associated with codeine (C) and investigated the possible molecular anchorage mechanisms of PA. Mice (n = 5 per group) were treated with PA alone and associated with codeine and assigned to the following groups: 75.0 mg/kg PA; 75.0 mg/kg PA + C 30 mg/kg; PA 37.5 mg/kg + C 15.0 mg/kg; C 30.0 mg/kg; and control. Nociception was induced by formalin, capsaicin, and glutamate, and was quantified based on the duration (in seconds) of face grooming. The possible mechanisms of action were evaluated by molecular docking study. In the formalin test, PA75/C30 presented an effect in the neurogenic (p < 0.0001) and inflammatory (p < 0.005) phases. Mice treated with PA75 (p < 0.0001) and PA75/C30 (p < 0.0005) showed a reduced nociceptive behavior in the capsaicin test. Glutamate-induced nociception also was blocked by PA75 (p < 0.0005) and C30 (p < 0.0005). The molecular anchorage analysis indicated high negative binding energy values for the evaluated receptors, especially glutamate receptors (AMPA -79.57 Kcal/mol, mGLUR6 -71.25, and NMDA -66.33 Kcal/mol). PA associated with codeine showed orofacial antinociceptive activity, with theoretical evidence of interaction with glutamate receptors.

Abstract We aimed to evaluate the orofacial antinociceptive effect of geraniol in mice and its molecular anchorage mechanism. Seven mice per group (probabilistic sample) were treated with geraniol (12.5, 25 and 50 mg/kg, i.p.), morphine (6 mg/kg, i.p.) and vehicle (saline + Tween 80 at 0.2%, i.p.) 30 minutes prior to the beginning of the experiment. Injecting glutamate (25 μM), capsaicin (2.5 μg) and formalin (2%) into the right upper lip (perinasal) of the mouse induced nociception. Behavioral analysis of the animals considered the friction time (in seconds) of the mentioned region using hind or front paws by a researcher blinded to the treatment groups. The statistical analysis was performed blindly, considering α = 5%. The results showed that in the glutamate and capsaicin tests, concentrations of 25 mg/kg and 50 mg/kg presented antinociceptive activity (p < 0.005, power> 80%). In the formalin test, geraniol was able to reduce nociception at a concentration of 50 mg/kg (p < 0.005, power> 80%). In the molecular anchorage study, high values of binding between the evaluated substance and receptors of glutamate were observed (metabotropic glutamate receptor, -87.8501 Kcal/mol; N-methyl-D-aspartate, -86.4451 Kcal/mol; α-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid, -85.6755 Kcal/mol). Geraniol presented orofacial antinociceptive activity, probably by interacting with glutamate-related receptors.

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.

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.

Os materiais de fosfato de cálcio, pelas características de biocompatibilidade e osteocondução, estão sendo estudados como substitutos ósseos. OBJETIVO: Comparar histomorfometricamente o reparo ósseo promovido pela cerâmica bifásica de hidroxiapatita, cimento fosfato de cálcio e enxerto ósseo autógeno. FORMA DE ESTUDO: Prospectivo e experimental em laboratório. MATERIAL E MÉTODO: Foram criadas duas cavidades (esquerda e direita - teste e controle) de 5mm de diâmetro nos ossos parietais de 72 ratos Wistar. As cavidades testes GI, GII, GIII e GIV foram preenchidas com cimento fosfato de cálcio, cerâmica bifásica de hidroxiapatita, enxerto ósseo autógeno e coágulo sanguíneo. Os animais foram sacrificados aos 30, 60 e 90 dias e os espécimes avaliados histomorfometricamente. RESULTADOS: Os resultados mostraram maior formação óssea no período de 30 dias para o GIII quando comparado aos outros grupos testes. Nos períodos de 60 e 90 dias, a neoformação óssea foi mais acentuada nos grupos testes GI, GII e GIII, alcançando maiores áreas em GI e GII. Em todos os períodos de avaliação, as cavidades testes GII apresentaram maiores áreas frente às GI. CONCLUSÃO: Os biomateriais foram superiores ao enxerto autógeno no reparo ósseo

Because of its biocompatibility and osteoconductive properties, calcium-phosphate cement has been used as bone surrogate. OBJECTIVE: The purpose of this study was to do a histomorphometric comparison of bone regeneration using hydroxyapatite biphasic ceramic, calcium-phosphate cement and autogenous bone graft. STUDY DESIGN: Prospective and laboratorial experiment. MATERIALS AND METHODS: Two 5mm in diameter cavities (left and right - test and control) were made in the parietal bone of 72 rats. The GI, GII, GIII and GIV test cavities were filled with calcium-phosphate cement, biphasic ceramic hydroxyapatite, autogenous bone graft and blood. The animals were killed at 30, 60 and 90 days and the specimens underwent histomorphometric analysis. RESULTS: The results showed that autogenous bone graft treated defects had significantly more new bone at 30 days compared to other test groups. Within 60 and 90 days, bone formation was more significant in the test groups GI, GII and GIII; GI and GII encompassed larger areas. Throughout the evaluation periods, GII tests showed more bone formation when compared to GI. CONCLUSIONS: Biomaterials depicted a significantly increase in bone content, when compared to autogenous bone graft, concerning bone regeneration

Trichophyton mentagrophytes é um fungo causador de dermatofitoses, afetando humanos em todo o mundo. Isto direciona a busca de produtos para o tratamento destas infecções. Assim, este estudo teve por objetivo investigar a atividade antifúngica do óleo essencial de Cymbopogon winterianus contra T. mentagrophytes. Os ensaios antifúngicos foram constituídos do screening antifúngico, da determinação CIM e CFM, da análise dos efeitos do óleo essencial no crescimento micelial, na germinação dos esporos, na viabilidade fúngica, na morfogênese, na parede celular (ensaio com sorbitol) e na membrana celular (ensaio de lise celular) de T. mentagrophytes. No screening, o óleo inibiu todas as cepas, com zonas de inibição de crescimento de 24-28 mm de diâmetro. A CIM foi de 312 μg/mL e a CFM foi de 2500 μg/mL para quase todas as cepas testadas. O óleo essencial inibiu o desenvolvimento micelial, a germinação dos esporos e a viabilidade fúngica. Houve alterações morfológicas no agrupamento dos conídios, na forma e pigmentação das hifas. A ação antifúngica do produto não envolve a parede celular e parece estar envolvida com a membrana celular fúngica. Pode-se concluir que o óleo essencial de C. winterianus se apresenta como um potencial produto antifúngico, especialmente para o tratamento das dermatofitoses.

Trichophyton mentagrophytes is a fungus causative agent of dermatophytosis, affecting humans worldwide. This has driven the search for products for the treatment of these infections. Accordingly, the aim of this study was to investigate the antifungal activity of the Cymbopogon winterianus essential oil against T. mentagrophytes. The antifungal tests consisted of antifungal screening, determination of MIC and MFC, analysis of the essential oil's effects on mycelial growth, germination of fungal spores, fungal viability, morphogenesis, cell wall (test with sorbitol) and cell membrane (cell leakage test) of T. mentagrophytes. Upon screening, the oil inhibited all strains, with zones of growth inhibition of 24-28 mm in diameter. The MIC was 312 μg/mL and CFM was 2500 μg/mL for almost all the strains tested. There were morphological changes in the conidia group, form and pigmentation of hyphae. The antifungal action of the product does not involve the cell wall and its action may involve the fungal plasma membrane. It is concluded that C. winterianus essential oil constitutes a potential antifungal product, especially for the treatment of dermatophytosis.

Trichophyton rubrum is one of the most common fungi causer of dermatophytosis, mycosis that affect humans and animals around the world. Researches aiming new products with antifungal activity become necessary to overcome difficulties on treatment of these infections. Accordingly, this study aimed to investigate the antifungal activity of essential oil from Cymbopogon winterianus against the dermatophyte T. rubrum. The antifungal screening was performed by solid medium diffusion method with 16 T. rubrum strains, minimum inhibitory concentration (MIC) and minimum fungicide concentration (MFC) were determined using the microdilution method. The effects on mycelial dry weight and morphology were also observed. Screening showed essential oil in natura inhibited all the tested strains, with inhibition zones between 24-28 mm diameter. MIC50 and MIC90 values of the essential oil were 312 µg/mL for nearly all the essayed strains (93.75 %) while the MFC50 and MFC90 values were about eight times higher than MIC for all tested strains. All tested essential oil concentrations managed to inhibit strongly the mycelium development. Main morphological changes on the fungal strains observed under light microscopy, which were provided by the essential oil include loss of conidiation, alterations concerning form and pigmentation of hyphae. In the oil presence, colonies showed folds, cream color and slightly darker than the control, pigment production was absent on the reverse and with evident folds. It is concluded that C. winterianus essential oil showed activity against T. rubrum. Therefore, it could be known as potential antifungal compound especially for protection against dermatophytosis.

O objetivo deste estudo foi observar a eficácia do óleo essencial de O. vulgare L. e O. majorana L. na inibição do crescimento e sobrevivência de cepas de fungos potencialmente patogênicas, bem como avaliar os possíveis mecanismos envolvidos no estabelecimento da propriedade antifúngica dos óleos essenciais testados através do ensaio de estabilidade osmótica e morfogênese. As cepas fúngicas utilizadas neste estudo foram Candida albicans ATCC 7645, C. tropicalis LM-14, C. krusei LM-09, Cryptococcus neoformans FGF-5, Aspergillus flavus LM-02, A. fumigatus IPP-21, T. rubrum ATCC 28184, T. mentagrophytes LM-64, Microsporum gypseum ATCC 184, M. canis LM-36 e Cladosporium herbarium ATCC 26362. O óleo essencial de O. vulgare apresentou valor de CIM de 80 µL/mL, enquanto o óleo essencial de O. majorana apresentou valor de CIM de 160 µL/mL. C. krusei LM-09 apresentou-se como a única cepa resistente a todas as concentrações ensaiadas de ambos os óleos essenciais. Os óleos essenciais testados quando ensaiadas em seu valor de CIM causaram um efeito fungicida contra C. albicans ATCC 7645 após 4 h de exposição. O óleo essencial de O. vulgare na concentração de 80 µL/mL exibiu uma total inibição do crescimento micelial radial de T. rubrum ATCC 28184 e M. canis LM-36 ao longo de 14 dias. As cepas fúngicas ensaiadas quando tratadas com sorbitol (agente osmo-protetor) foram capazes de crescer em meio adicionado de mais altas concentrações dos óleos essenciais quando comparados ao meio não adicionado de sorbitol, sugerindo especificidade destes produtos a parede celular como alvo na célula fúngica. As principais alterações causadas pelo óleo essencial de O. vulgare sobre a morfologia de A. flavus LM-02 foram diminuída conidiação, perda de citoplasma, perda de pigmentação e ruptura da estrutura celular indicando degeneração da parede celular fúngica. Estes resultados sugerem que óleos essenciais de espécies de Origanum poderiam ser considerados como potenciais antifúngicos para o controle de fungos patógenos e ocorrência de micoses.

This study aimed to assess the efficacy of O. vulgare L. and O. majorana L. essential oil in inhibiting the growth and survival of potentially pathogenic fungal strains and also sought to evaluate the possible mechanisms involved in the establishment of the antifungal property of the tested essential oils through assays of osmotic stability and morphogenesis. Test strains included in this study were Candida albicans ATCC 7645, C. tropicalis LM-14, C. krusei LM-09, Cryptococcus neoformans FGF-5, Aspergillus flavus LM-02, A. fumigatus IPP-21, T. rubrum ATCC 28184, T. mentagrophytes LM-64, Microsporum gypseum ATCC 184, M. canis LM-36 and Cladosporium herbarium ATCC 26362. O. vulgare essential oil presented a MIC value of 80 µL/mL, while for O. majorana this was 160 µL/mL. C. krusei LM-09 was the only strain resistant to all assayed concentrations of both essential oils. O. vulgare and O. majorana essential oil at their MIC values provided a cidal effect against C. albicans ATCC 7645 after 4 h of exposure. O. vulgare essential oil at 80 µL/mL exhibited 100 % inhibition of the radial mycelia growth of T. rubrum ATCC 28184 and M. canis LM-36 for 14 days. Assayed fungus strain protected by sorbitol (osmo-protectant agent) grew in media containing higher concentrations of O. vulgare and O. majorana essential oil in comparison to media without sorbitol, suggesting some specificity of these essential oils for targeting cell wall in the fungi cell. Main morphological changes observed under light microscopy provided by the essential oil of O. vulgare in A. flavus LM-02 were decreased conidiation, leakage of cytoplasm, loss of pigmentation and disrupted cell structure indicating fungal wall degeneration. These results suggest that essential oils from Origanum could be regarded as a potential antifungal compound for controlling the growth of pathogen fungi and the occurrence of mycoses.

Cinnamomum zeylanicum Blume é uma planta conhecida por apresentar ampla variedade de propriedades medicinais. Portanto, este estudo teve por objetivo avaliar a interferência do óleo essencial C. zeylanicum sobre o crescimento e morfogênese de algumas espécies de Aspergillus potencialmente patogênicas. O óleo essencial testado apresentou potente efeito antifúngico demonstrado pela visualização de grandes zonas de inibição de crescimento de todas as linhagens testadas. Os valores de CIM50 e de CIM90 foram 40 e 80 µL/mL, respectivamente. Nas concentrações de 80, 40 e 20 µL/mL o óleo demonstrou um potente efeito fumigante, inibindo o crescimento micelial radial de A. niger, A. flavus e A. fumigatus ao longo de 14 dias de exposição. A 80 e 40 µL/mL o óleo essencial promoveu inibição de 100% da germinação de esporos, das três espécies de Aspergillus citadas anteriormente. Além disso, alterações morfológicas no crescimento fúngico foram observadas sob microscopia óptica após exposição ao óleo essencial, como diminuição da conidiação, perda citoplasmática, perda de pigmentação e rompimento da estrutura fúngica (hifa) indicando degeneração da parede celular. Diante do exposto, conclui-se que o óleo essencial de C. zeylanicum poderia ser empregado como potente composto antifúngico, particularmente, prevenindo crescimento de espécies de Aspergillus.

Cinnamomum zeylanicum Blume is known for a wide range of medicinal properties. This study aimed to assess the interference of C. zeylanicum essential oil on the growth and morphogenesis of some potentially pathogenic Aspergillus species. The essential oil presented strong antifungal effect causing the growth inhibition of the assayed strains and development of large growth inhibition zones. MIC50 and MIC90 values were 40 and 80 µL/mL, respectively. 80, 40 and 20 µL/mL of the oil strongly inhibited the radial mycelial growth of A. niger, A. flavus and A. fumigatus along 14 days. 80 and 40 µL/mL of the oil caused a 100% inhibition of the fungal spore germination. Main morphological changes observed under light microscopy provided by the essential oil in the fungal strains were decreased conidiation, leakage of cytoplasm, loss of pigmentation and disrupted cell structure indicating fungal wall degeneration. It is concluded that C. zeylanicum essential oil could be known as potential antifungal compound, particularly, to protect against the growth of Aspergillus species.

As análises bidimensionais no estudo histopatológico de lesões cariosas apresentam vieses que podem comprometer o valor científico dos estudos. Neste artigo, os autores apresentam uma nova técnica de análise tridimensional de lesões cariosas, sem perda de partes da lesão, que consiste na dissecação das áreas afetadas pela lesão com brocas odontológicas de desgaste, sob refrigeração, realizada com auxílio de um estereomicroscópio. Três casos de lesões cariosas oclusais em primeiros e segundos molares permanentes são apresentados, nos quais foi possível observar as reações internas de esmalte e dentina em relação às características macroscópicas das lesões. Alguns aspectos comparativos entre a técnica da dissecação e as técnicas bidimensionais são discutidos.

Two-dimensional analyses of the histopathological aspect of carious lesions present bias that may compromise the scientific value of studies. In this paper, the authors present a new three-dimensional technique for the histopathological analysis of carious lesions. It consists in the dissection of the affected areas under the stereomicroscope, utilizing refrigerated dental burs, without any loss of carious tissue. The dissection of three occlusal lesions located on first and second permanent molars is presented. It was possible to observe the internal reactions of enamel and dentin, and correlate the results to the macroscopic features of the occlusal surface. Some comparative aspects between the dissecting and two-dimensional techniques are discussed.