Effect of green Coffea arabica L. seed oil on extracellular matrix components and water-channel expression in in vitro and ex vivo human skin models

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Microbiological or Chemical Models of Enamel Secondary Caries Compared by Polarized-Light Microscopy and Energy Dispersive X-ray Spectroscopy

Different secondary caries models may present different results. The purpose of this study was to compare different in vitro secondary caries models, evaluating the obtained results by polarized-light microscopy (PLM), scanning electron microscopy (SEM), and energy dispersive X-ray spectroscopy (EDS). Standardized human enamel specimens (n = 12) restored with different materials (Z250 conventional composite resin-CRZ, Freedom polyacid-modified composite resin-CRF, Vitremer resin-modified glass-ionomer-GIV, and Fuji IX conventional glass-ionomer cement-GIF) were submitted to microbiological (MM) or chemical caries models (CM). The control group was not submitted to any caries model. For MM, specimens were immersed firstly in sucrose broth inoculated with Streptococcus mutans ATCC 35688, incubated at 37 degrees C/5% CO(2) for 14 days and then in remineralizing solution for 14 days. For CM, specimens were submitted to chemical pH-cycling. Specimens were ground, submitted to PLM and then were dehydrated, gold-sputtered and submitted to SEM and EDS. Results were statistically analyzed by Kruskall-Wallis and Student-Newman-Keuls tests (alpha = 0.05). No differences between in vitro caries models were found. Morphological differences in enamel demineralization were found between composite resin and polyacid-modified composite resin (CRZ and CRF) and between the resin-modified glass-ionomer and the glass-ionomer cement (GIF and GIV). GIF showed higher calcium concentration and less demineralization, differing from the other materials. In conclusion, the glass-ionomer cement showed less caries formation under both in vitro caries models evaluated. (C) 2009 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater 90B: 635-640, 2009

Efeito cariostático de restaurações adesivas em superfícies radiculares: estudo in vitro

Materiais restauradores que liberam íons flúor e/ou promovem adesão à estrutura dental têm sido relacionados com a inibição do desenvolvimento de lesões de cárie adjacentes às restaurações. A hipótese testada neste estudo foi a de que o uso de resina composta/sistema adesivo tem efeito cariostático semelhante a um material adesivo que libera íons flúor - cimento de ionômero de vidro - sobre a superfície radicular adjacente às restaurações. Foram utilizadas 20 raízes de terceiros molares humanos extraídos, embutidas em resina de poliestireno e planificadas. Cavidades padronizadas foram preparadas e restauradas aleatoriamente com (a) Chelon-Fil (Espe) ou (b) Z100/SingleBond (3M). Valores iniciais (KHNi) de microdureza superficial Knoop da dentina foram obtidos a 100, 200 e 300 mim da margem oclusal das restaurações. Uma área de 2,0 mm ao redor da restauração foi delimitada e submetida à indução de cárie artificial. Obtiveram-se, então, os valores finais (KHNf) de microdureza, nas mesmas condições e localizações da leitura inicial. As diferenças entre KHNi e KHNf foram consideradas para a análise estatística. As medianas de KHNi - KHNf nas distâncias de 100, 200 e 300 mim foram para (a): -3,8; -0,3; -1,0; e para (b): 3,3; 2,5; 1,7. O teste de Kruskal-Wallis não evidenciou diferença significativa entre as distâncias dentro de cada grupo. Às distâncias de 200 e 300 mim, não houve diferença significativa entre os materiais avaliados. À distância de 100 mim, (a) diferiu significativamente de (b) (p < 0,05). Sob as condições deste estudo, o cimento de ionômero de vidro apresentou maior potencial cariostático que a resina composta com sistema adesivo dentinário.

In vitro evaluation of acidified toothpastes with low fluoride content

Fluoride toothpastes are a risk factor for the development of dental fluorosis. Products with low fluoride content offer a higher security, but their effectiveness must be proven. The aim of this in vitro study was to compare two acidified toothpastes with low fluoride concentration (412 and 550 mu g F/g) with neutral toothpastes. Bovine enamel blocks were selected by surface microhardness (SMH) and randomized to twelve groups of 13, according to the fluoride concentration in toothpaste (placebo, 275, 412, 550 or 1,100 jig F/g) and pH (7.0 or 5.5). Two commercially available toothpastes were also studied: a 1,100-mu g F/g, pH 7.0 paste (positive control) and a children's paste (500 mu g F/g, pH 7.0). The blocks were subjected to pH cycling for 7 days. The toothpaste treatment was done twice daily. Surface and cross-sectional microhardnesses were assessed to calculate the percentage change of SMH (%SMH) and the mineral loss (Delta Z). The amount of fluoride, calcium and phosphorus in the solutions after the pH cycling was also analyzed. Compared to neutral toothpastes, the acidified toothpastes reduced the %SMH in all F concentrations. Higher F and lower Ca and P concentrations were found in solutions for the acidified toothpastes. Regarding AZ, only the positive control, 1,100-mu g F/g (acidified and neutral) groups were not statistically different. The acidified toothpastes showed a dose-response relationship with all variables. For the low-fluoride toothpastes evaluated, only the 550-mu g F/g acidified paste had the same anticariogenic action as the 1,100-mu g F/g neutral paste.

In vitro evaluation of the effectiveness of acidic fluoride dentifrices

This study evaluated the effectiveness of acidic low-fluoride dentifrices compared to conventional neutral dentifrices. Enamel blocks were submitted to pH cycling and treatment with slurries of dentifrices containing 0, 275, 412, 550 and 1,100 mu g F/g (pH 4.5 or 7.0), and also a commercial dentifrice (1,100 mu g F/g) and a commercial children's dentifrice (500 mu g F/ g). Variations in surface microhardness and in the mineral content in enamel after pH cycling were calculated. Enamel blocks treated with acidic dentifrices exhibited less mineral loss compared to neutral dentifrices (ANOVA; p < 0.05). The acidic dentifrices with 412 and 550 mu g F/g had the same effectiveness as the neutral 1,100-mu g F/g dentifrice and commercial 1,100-mu g F/g dentifrice. Copyright (c) 2007 S. Karger AG, Basel

Inhibition of demineralization in vitro around amalgam restorations

Objective: The purpose of this in vitro study was to evaluate some forms of preventing or avoiding demineralization within enamel cavity walls adjacent to amalgam restorations. Method and materials: Third molar teeth were sectioned to obtain 72 specimens, divided into one control and five experimental groups: amalgam only; varnish plus amalgam; acidulated phosphate fluoride plus amalgam; adhesive amalgam; glass-ionomer cement plus amalgam; control (amalgam only, not subjected to a demineralization challenge). The experimental groups were subjected to pH and thermal cycling and then submitted to enamel hardness determinations. Results: Significant differences between the treatment groups revealed that the bonded amalgam technique offered the best resistance to demineralization. The use of cavity varnish resulted in greater mineral loss than amalgam placed alone. Conclusion: The use of an adhesive system, glass-ionomer cement, or acidulated phosphate fluoride under amalgam restorations may interfere with development of secondary caries.

In vitro-evaluation of secondary caries formation around restoration.

The objective of this in vitro study was to evaluate demineralization around restorations. Class V preparations were made on the buccal and lingual surfaces of each tooth. TPH (Group 1), Fuji II LC (Group 2), Tetric (Group 3), Dyract (Group 4), GS 80 (Group 5) and Chelon Fil (Group 6) were randomly placed in equal numbers of teeth. The teeth were submitted to a pH-cycling model associated with a thermocycling model. Sections were made and the specimens were examined for the presence of demineralization under polarized light microscopy. Demineralization was significantly reduced with Chelon Fil (Group 6). Furthermore, a similar inhibitory effect on the development of demineralization was observed in Groups 2, 4 and 5.

In vitro susceptibility of oral Candida albicans strains to different pH levels and calcium hydroxide saturated aqueous solution

Candida albicans is present in the oral cavity and in the whole digestive tract of humans and other animals, being frequently related to endodontic treatment failure. The present study determined the incidence of C. albicans in the oral cavity and the susceptibility of isolates to different pH values and saturated calcium hydroxide aqueous solution at pH 12.5. Sixty-five patients attending the Endodontic Clinic at the Sagrado Coração University participated in the study. The collected samples were cultivated in selective media for C. albicans and the isolates were tested in terms of resistance to both alkaline pH and saturated aqueous solution of calcium hydroxide. In relation to time variables, yeast viability was assessed by the Sabouraud's agar culture and fluorescein diacetate and ethidium bromide fluorescent staining method. Results from the different pHs and experimental times, including those from different techniques measuring fungal viability, were compared using the chi-square and Fisher's exact tests (α=0.05). The yeasts became completely inviable after 48 h of contact with the calcium hydroxide solution. On the other hand, when exposed to the alkaline culture broth, the yeasts were found to be viable at pHs 9.5 and 10.5 for up to 7 days. In conclusion, C. albicans can only be completely inhibited by direct contact with saturated calcium hydroxide aqueous solution after 48 h of exposure.

Antifungal Efficacy during Candida krusei Infection in Non-Conventional Models Correlates with the Yeast In Vitro Susceptibility Profile

The incidence of opportunistic fungal infections has increased in recent decades due to the growing proportion of immunocompromised patients in our society. Candida krusei has been described as a causative agent of disseminated fungal infections in susceptible patients. Although its prevalence remains low among yeast infections (2-5%), its intrinsic resistance to fluconazole makes this yeast important from epidemiologic aspects. Non mammalian organisms are feasible models to study fungal virulence and drug efficacy. In this work we have used the lepidopteran Galleria mellonella and the nematode Caenorhabditis elegans as models to assess antifungal efficacy during infection by C. krusei. This yeast killed G. mellonella at 25, 30 and 37°C and reduced haemocytic density. Infected larvae melanized in a dose-dependent manner. Fluconazole did not protect against C. krusei infection, in contrast to amphotericin B, voriconazole or caspofungin. However, the doses of these antifungals required to obtain larvae protection were always higher during C. krusei infection than during C. albicans infection. Similar results were found in the model host C. elegans. Our work demonstrates that non mammalian models are useful tools to investigate in vivo antifungal efficacy and virulence of C. krusei. © 2013 Scorzoni et al.

Anti-inflammatory mechanisms of the annexin A1 protein and its mimetic peptide Ac2-26 in models of ocular inflammation in vivo and in vitro

Annexin A1 (AnxA1) is a protein that displays potent anti-inflammatory properties, but its expression in eye tissue and its role in ocular inflammatory diseases have not been well studied. We investigated the mechanism of action and potential uses of AnxA1 and its mimetic peptide (Ac2-26) in the endotoxin-induced uveitis (EIU) rodent model and in human ARPE-19 cells activated by LPS. In rats, analysis of untreated EIU after 24 and 48 h or EIU treated with topical applications or with a single s.c. injection of Ac2-26 revealed the anti-inflammatory actions of Ac2-26 on leukocyte infiltration and on the release of inflammatory mediators; the systemic administration of Boc2, a formylated peptide receptor (fpr) antagonist, abrogated the peptide's protective effects. Moreover, AnxA1-/- mice exhibited exacerbated EIU compared with wild-type animals. Immunohistochemical studies of ocular tissue showed a specific AnxA1 posttranslational modification in EIU and indicated that the fpr2 receptor mediated the anti-inflammatory actions of AnxA1. In vitro studies confirmed the roles of AnxA1 and fpr2 and the protective effects of Ac2-26 on the release of chemical mediators in ARPE-19 cells. Molecular analysis of NF-κB translocation and IL-6, IL-8, and cyclooxygenase-2 gene expression indicated that the protective effects of AnxA1 occur independently of the NF-κB signaling pathway and possibly in a posttranscriptional manner. Together, our data highlight the role of AnxA1 in ocular inflammation, especially uveitis, and suggest the use of AnxA1 or its mimetic peptide Ac2-26 as a therapeutic approach. Copyright © 2013 by The American Association of Immunologists, Inc.

Effect of sodium trimetaphosphate on hydroxyapatite solubility: An in vitro study

This study evaluated the effect of different concentrations of sodium trimetaphosphate (TMP) with and without fluoride (F) on the concentration of calcium (Ca), phosphorus (P) and F in hydroxyapatite (HA). Synthetic HA powder (0.15 g) was suspended (n=6) in solutions (75 mL) of TMP at 0%, 0.1%, 0.2%, 0.4%, 0.6%, 0.8%, 1.0%, 2.0%, 4.0%, 6.0%, 8.0% and 10% concentrations in the presence and absence of 100 ppm F and subjected to a pH-cycling process. The precipitates were filtrated, dried at 70° C for 24 h and ground onto a fine powder. The concentrations of F (KOH (CaF2) and HCl (FA) soluble), Ca (Arsenazo III), and P (molybdate method) in HA were determined. The Ca P, and Ca/P ratio data were subjected to Tukey's test and the F data were subjected to Student-Newman-Keuls test (p<0.05). The addition of TMP to the samples reduced F deposition to 98% (p<0.001). The groups containing 100 ppm F and 0.4% or 0.6% TMP exhibited a higher Ca concentration than the group containing only 100 ppm F (p<0.05). Furthermore, the HA treated with 0.2% and 0.4% TMP and 100 ppm F showed a higher Ca/P ratio than the other groups (p<0.001). In conclusion, TMP at 0.2%, 0.4% and 0.6% concentrations combined with F seemed to be able to precipitate HA with low solubility. However, especially at high concentrations, TMP interferes with F deposition on HA.

In vitro enamel remineralization by low-fluoride toothpaste with calcium citrate and sodium trimetaphosphate

The objective of this study was to evaluate in vitro the effect of a low fluoride toothpaste (450 μgF/g, NaF) combined with calcium citrate (Cacit) and sodium trimetaphosphate (TMP) on enamel remineralization. Bovine enamel blocks had the enamel surface polished sequentially to determine the surface hardness. After production of artificial carious lesions, the blocks selected by their surface hardness were submitted to remineralization pH cycling and daily treatment with dentifrice suspensions (diluted in deionized water or artificial saliva): placebo, 275, 450, 550 and 1,100 μgF/g and commercial dentifrice (positive control, 1,100 μgF/g). Finally, the surface and cross-section hardness was determined for calculating the change of surface hardness (%SH) and mineral content (%ΔZ). Fluoride in enamel was also determined. The data from %SH, %ΔZ and fluoride were subjected to two-way analysis of variance followed by Student-Newman-Keuls's test (p<0.05). The mineral gain (%SH and %ΔZ) was higher for toothpastes diluted in saliva (p<0.05), except for the 450 mgF/g dentifrice with Cacit/TMP (p>0.05). The 450 Cacit/ TMP toothpaste and the positive control showed similar results (p>0.05) when diluted in water. A dose-response was observed between fluoride concentration in toothpastes and fluoride present in enamel, regardless of dilution. It was concluded that it is possible to enhance the remineralization capacity of low F concentration toothpaste by of organic (Cacit) and inorganic (TMP) compounds with affinity to hydroxyapatite.

In vitro evaluation of the effect of mouth rinse with trimetaphosphate on enamel demineralization

Objective: The aim of this study was to investigate the effectiveness of sodium trimetaphosphate (TMP) addition to mouth rinses to inhibit enamel demineralization. Design: Bovine enamel blocks (n = 88) were selected by surface hardness and divided into eight treatment groups (n = 11 per group): placebo, 100 or 225 μg F/ml; the rinses with 100 μg F/ml had differing TMP concentrations (range 0-0.6%). The blocks were subjected to pH cycling for 5 days and treated twice a day with mouth rinses. After that, surface and cross-sectional hardness as well as fluoride in enamel were measured. Results: The groups containing both 100 μg F/ml and 0.4% TMP inhibited demineralization most effectively (p < 0.001). This formulation yielded lower values of lesion areas than the formulations containing 100 or 225 μg F/ml but no TMP. The addition of 0.4% TMP increased the fluoride in enamel. Conclusion: It is possible to improve the effectiveness of a mouth rinse with 100 μg F/ml by addition of TMP, this being superior in inhibiting enamel demineralization compared with mouth rinses containing 225 μg F/ml. © 2013 S. Karger AG, Basel.