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)

Combined treatment with caffeic and ferulic acid from Baccharis uncinella C. DC. (Asteraceae) protects against metabolic syndrome in mice

Fractionation of the EtOH extract from aerial parts of Baccharis uncinella C. DC. (Asteraceae) led to isolation of caffeic and ferulic acids, which were identified from spectroscopic and spectrometric evidence. These compounds exhibit antioxidant and anti-inflammatory properties and have been shown to be effective in the prevention/treatment of metabolic syndrome. This study investigated whether the combined treatment of caffeic and ferulic acids exhibits a more significant beneficial effect in a mouse model with metabolic syndrome. The combination treatment with caffeic and ferulic acids was tested for 60 days in C57 mice kept on a high-fat (40%) diet. The data obtained indicated that treatment with caffeic and ferulic acids prevented gain in body weight induced by the high-fat diet and improved hyperglycemia, hypercholesterolemia and hypertriglyceridemia. The expression of a number of metabolically relevant genes was affected in the liver of these animals, showing that caffeic and ferulic acid treatment results in increased cholesterol uptake and reduced hepatic triglyceride synthesis in the liver, which is a likely explanation for the prevention of hepatic steatosis. In conclusion, the combined treatment of caffeic and ferulic acids displayed major positive effects towards prevention of multiple aspects of the metabolic syndrome and liver steatosis in an obese mouse model.

Effect of green tea as a protective measure against dental erosion in coronary dentine

Abstract The aim of this study was to evaluate the effect of green tea as a protective measure on eroded dentin. Disks of human coronary dentin were selected based on surface hardness and randomly assigned to 3 groups (n = 10): DW - distilled water, CHX - 0.2% chlorhexidine digluconate, and GT - green tea. The disks were allowed to acquire pellicle for 2 hours and were then subjected to 3 cycles per day of demineralization (C6H8O7 0.05 M, pH 3.75, 60 s), treatment (DW or CHX or GT, 5 min) and remineralization (artificial saliva, 60 min) over a period of 3 days. Changes in the dentin were determined by loss of surface hardness (%SHL) and mechanical profilometry analysis at the end of each day. Data were analyzed by two-way ANOVA followed by Tukey’s test for %SHL and profilometry (p < 0.05). Significant reductions in dentin hardness loss were observed only for the CHX group when compared to the DW group (p < 0.05). However, there was no significant difference between the CHX and GT groups (p > 0.05). A significant difference was observed between DW and GT treatments for wear and roughness measurements (p < 0.05). The green tea extract solution was able to reduce the wear and roughness caused by dentin erosion under the conditions of this study.

Protective effect of Dillenia indica L. on acetic acid induced colitis in mice.

The inflammatory bowel disease (IBD) is an idiopathic, immune mediated and chronic inflammation of the intestine. The study aimed to elucidate the ameliorative effect of methanolic extract of Dillenia indica (DIME), hexane fraction (HFDI) and chloroform fraction (CFDI) of Dillenia indica in acetic acid induced experimental colitis in mice. Macroscopic score, colon weight, colonic catalase (CAT), superoxide dismutase (SOD), glutathione (GSH), myeloperoxidase (MPO), malondialdehyde (MDA), tumor necrosis factor (TNF-α), and histological changes were recorded after the treatment regimen of 7 days. Intra-rectal instillation of acetic acid caused enhanced macroscopic score, colon weight, colonic MPO, MDA, and TNF-α level. It caused significant decreased level of CAT, SOD and GSH. DIME (800 mg/kg), HFDI (200 mg/kg) and CFDI (200 mg/kg) treatment exhibited significant effect in lowering macroscopic score, colon weight, MPO, MDA, TNF-α levels and elevation of CAT, GSH and SOD levels. The results suggest that D. indica has ameliorating effects on experimental colitis by inhibiting the proinflammatory mediators like TNF-α production.

Prophylactic effect of Mucuna pruriens Linn (velvet bean) seed extract against experimental Naja sputatrix envenomation: Gene expression studies.

Mucuna pruriens is widely used in traditional medicine for treatments of various diseases. In certain region of Nigeria, the seed is used as oral prophylactics for snakebite. Rats pretreated with the aqueous extract from M. pruriens seed (MPE) were protected against the lethal effects of Naja sputatrix (Javan spitting cobra) venom [Tan et al., J Ethnopharmacol, 123 (2009) 356]. The pretreatment also protected against venom-induced histopathological changes in rat heart. To contribute to the understanding of the mechanism of cardio-protective action, the present study examined the effects of MPE-pretreatment on gene expression profile of rat heart as well as effect of MPE-pretreatment on N. sputatrix venom-induced gene expression alterations in rat heart. The gene expression profiles were examined by microarray analysis and verified by real time PCR. The results showed that pretreatment with MPE caused 50 genes in the rat heart substantially up-regulated of which 19 were related to immune responses, 7 were related to energy production and metabolism. The up-regulation of genes related to energy metabolism probably plays a role in maintaining the viability of the heart. Four other genes that were up-regulated (alpha synuclein, natriuretic peptide precursor, calsequestrin and triadin) were involved in the maintenance of homeostasis of the heart or maintaining its viability, thereby contributing to the direct protective action. The results demonstrated that protective effect of MPE pretreatment against snake venom poisoning may involve a direct action on the heart.

CCl[4] ilnduced hepatotoxicity: protective effects of carnosine on peroxisome proliferator-activated receptor gamma gene expression, genotoxicity and oxidant/antioxidant status in rats

Effect of carnosine as an antioxidant in protection against carbon tetrachloride CCI[4] induced oxidative stress and hepatotoxicity in rats was investigated. Liver toxicity was induced in rat model at which four experimental groups of 20 rats each were constructed: group [1] the control group in which rats were not administrated CCI[4] or carnosine; group [II] CCI[4] group in which rats were subcutaneously injected with CCI[4] in a dose of 2 ml /Kg body weight twice weekly for 4 weeks; group[III] CCI[4] and carnosine group in which rats were also subcutaneously injected with CCI[4] and co-treated with daily intraperitoneal [i.p.] carnosine at a dose of 10 mg / kg body weight and group [IV] received also i.p. repeated daily dose of carnosine. Hepatotoxicity was assessed by measurement of serum aspartate aminotransferase [AST] and alanine aminotransferase [ALT] activities. Hepatic peroxisome proliferator-activated receptor gamma [PPARgammay] mRNA expression, glutathione-S-transferase [GST] activity, paraoxonase 1 [PON1] activity, xantheine oxidase [XO] activity and total anti-oxidant capacity [TAC] level as well as DNA damage in blood were evaluated. The results were confirmed by histopathological examination. Carnosine treatment significantly prevented the CCI[4] induced hepatotoxicity, oxidative stress and DNA damage. In conclusion, our results suggested that carnosine might be a therapeutic antifibrotic/antigenotoxic agent for the treatment of CCI[4-] induced hepatotoxicity due to its antioxidant properties

The influence of protective varnish on the integrity of orthodontic cements

OBJECTIVE: The aim of the present study was to assess the influence of saliva contamination over the structural strength and integrity of conventional glass-ionomer cements used for cementing orthodontic bands in the absence and presence of a surface-protecting varnish. METHOD: 48 samples were prepared by inserting 3 types of glass-ionomer cements into standardized metallic matrixes of 10 mm of diameter and 2 mm of depth. The cements used were: Meron (VOCO), Ketac-Cem (3M ESPE) and Vidrion C (DFL), all of which comprised groups A, B and C, respectively. Subgroups A1, B1 and C1 comprised samples with no surface protection, whereas subgroups A2, B2 and C2 comprised samples of which surface was coated with Cavitine varnish (SS White), after cement manipulation and application, in order to protect the cement applied. All samples were stored in artificial saliva for 24 hours at 37ºC. A Vickers diamond micro-durometer was used to produce indentations on the non-treated group (non-varnished) and the treated group (varnished). RESULTS: Varnished materials had significantly higher microhardness values in comparison to non-varnished materials. Ketac-Cem had the highest microhardness value among the varnished materials. CONCLUSION: Varnish application is necessary to preserve the cement and avoid enamel decalcification. Glass-ionomer cements should be protected in order to fully keep their properties, thus, contributing to dental health during orthodontic treatment.

OBJETIVO: avaliar a influência da contaminação salivar na resistência estrutural e integridade de cimentos de ionômero de vidro convencionais utilizados para cimentação de bandas ortodônticas na ausência e na presença de um verniz protetor de superfície. MÉTODOS: quarenta e oito corpos de prova foram confeccionados a partir de três cimentos ortodônticos, com auxílio de matrizes metálicas padronizadas com 10mm de diâmetro e 2mm de altura. Os cimentos utilizados foram: Meron (Voco), Ketac-Cem (3M ESPE) e Vidrion C (DFL), compondo os grupos A, B e C, respectivamente. Metade dessas amostras não recebeu nenhum tipo de proteção superficial, constituindo os subgrupos A1, B1 e C1, enquanto, os subgrupos A2, B2 e C2 tiveram suas superfícies isoladas com verniz Cavitine (SS White) após manipulação e aplicação do cimento, com intuito de proteger a superfície do cimento. As amostras foram armazenadas em saliva artificial por 24 horas a 37ºC. Foi realizado um ensaio de microdureza (Vickers) para avaliação da dureza de superfície do grupo não-tratado (sem isolamento) e do grupo tratado (agente protetor). RESULTADOS: os materiais previamente isolados com o verniz obtiveram valores de microdureza significativamente maiores que os não-isolados. O cimento Ketac-Cem apresentou, estatisticamente, a maior microdureza entre os materiais protegidos. CONCLUSÃO: o isolamento com verniz mostrou-se necessário para preservação do cimento e, consequentemente, de sua capacidade de evitar possíveis desmineralizações dentárias. Os cimentos de ionômero de vidro devem ser protegidos para manutenção de sua integridade, contribuindo para saúde dental durante o tratamento ortodôntico.

El propóleos reduce la esteatosis hepática inducida por dieta en ratones / Propolis decrease diet-induced hepatic steatosis in mice

El hígado graso no alcohólico (HGNA), comprende un amplio espectro de lesiones, que van desde esteatosis hepática, hasta cirrosis y carcinoma hepatocelular, siendo sus principales factores de riesgo los desordenes asociados a síndrome metabólico (SM). El propóleos, sustancias resinosa elaborada por Apis mellifera para la protección de la colmena, ha demostrado un efecto hepatoprotector, así el objetivo de esta investigación fue evaluar el efecto de un propóleos chileno sobre el desarrollo de esteatosis hepática no alcohólica en ratones C57BL/6J sometidos a una dieta aterogénica tipo Paigen. Veintiocho ratones (C57BL/6J), divididos en: 1, dieta balanceada (CH); 2, dieta hipercolesterolémica (HC); 3, dieta HC + 10 mg/kg/día de propóleos (BP); 4, dieta HC + 40 mg/kg/día de propóleos (AP). Después de 16 semanas, se determinó la concentración de glucosa, colesterol total, triglicéridos y los niveles de alanina aminotransferasa (ALAT). El tejido hepático fue fijado en una solución de formalina tamponada al 10 por ciento para, posteriormente, ser incluido en paraplast y teñido con Hematoxilina-Eosina y fragmanetos congelados a -30C fijados en formalina teñidos con tinción Oil red O. No existieron diferencias en la concentración de glucosa ni triglicéridos, a diferencia del colesterol total, entre el grupo alimentado con dieta balanceada (CH) y los alimentados con la dieta aterogénica D12336 (HC, BP y AP). De la misma forma se observó que existen diferencias en los niveles de actividad de ALAT entre los grupos estudiados, destacando su reducción en los grupos suplementados con propóleos (BP y AP). De forma concordante, la histoarquitectura del grupo HC, mostró esteatosis simple y focos de infiltrado leucocitario en el lobulillo hepático, observándose una disminución gradual de las alteraciones en los grupos de BP y AP. En conclusión, el propóleos chileno estudiado disminuye la esteatosis hepática inducida por la dieta aterogénica tipo Paigen en ratones C57BL/6J, sin embargo, los mecanismos implicados en esta actividad hepatoprotectora tienen que ser identificados.

Nonalcoholic fatty liver disease (NAFLD) covers a wide spectrum of injuries ranging from simple steatosis to cirrhosis and hepatocellular carcinoma. Its main risk factors are disorders associated with metabolic syndrome (MS). Propolis, a resinous substance produced by Apis mellifera to protect is hive, has demonstrated a hepatoprotective effect. Thus, the aim of this study was to evaluate the effect of Chilean propolis on development of nonalcoholic hepatic steatosis in C57BL/6J mice exposed to Paigen atherogenic diet. Twenty eight mice C57BL/6J were divided four groups: 1, balanced diet (CH); 2, hypercholesterolemic diet (HD); 3, HD diet supplemented with 10 mg/kg/day of propolis (LP); 4, HD diet supplemented with 40 mg/kg/day of propolis (HP). After 16 weeks of treatment was determined glucose, total cholesterol and triglycerides concentrations and alanine aminotransferase (ALT) activity. The liver tissue was fixed in 10 percent buffered formalin solution, embedded in paraplast and stained with hematoxylin-eosin. No differences was detected in glucose and triglycerides concentrations, contrasting with total cholesterol levels between group fed with a balanced diet (CH) and feed with atherogenic diet D12336 (HD, LP and HP). In the same way, it was noted differences in ALT activity between groups, standing out the reduction in propolis supplemented groups (LP and HP). Likewise the histoarchitecture of HD group, showed simple steatosis, inflammatory cell infiltration and inflammatory foci in hepatic lobule. This characteristics show a gradual decrease in LP and HP groups. In conclusion, the Chilean propolis analyzed decreases hepatic steatosis induced by atherogenic diet in C57BL/6J mice. However, the involved mechanisms in this hepatoprotective activity must be identified.