Analysis by confocal laser scanning microscopy of the MDPB bactericidal effect on S. mutans biofilm CLSM analysis of MDPB bactericidal effect on biofilm.

UNLABELLED: Since bacteria remain in the dentin following caries removal, restorative materials with antibacterial properties are desirable to help maintaining the residual microorganisms inactive. The adhesive system Clearfil Protect Bond (PB) contains the antibacterial monomer 12-methacryloyloxydodecylpyridinium bromide (MDPB) in its primer, which has shown antimicrobial activity. However, its bactericidal effect against biofilm on the dentin has been little investigated. OBJECTIVE: The aim of this study was to analyze by confocal laser scanning microscopy (CLSM) and viable bacteria counting (CFU) the MDPB bactericidal effect against S. mutans biofilm on the dentin surface. MATERIAL AND METHODS: Bovine dentin surfaces were obtained and subjected to S. mutans biofilm formation in BHI broth supplemented with 1% (w/v) sucrose for 18 h. Samples were divided into three groups, according to the primer application (n=3): Clearfil Protect Bond (PB), Clearfil SE Bond, which does not contain MDPB, (SE) and saline (control group). After the biofilm formation, Live/ Dead stain was applied directly to the surface of each sample. Next, 10 µL of each primer were applied on the samples during 590 s for the real-time CLSM analysis. The experiment was conducted in triplicate. The primers and saline were also applied on the other dentin samples during 20, 90, 300 and 590 s (n=9 for each group and period evaluated) and the CFU were assessed by colonies counting. RESULTS: The results of the CLSM showed that with the Se application, although non-viable bacteria were detected at 20 s, there was no increase in their count during 590 s. In contrast, after the PB application there was a gradual increase of non-viable bacteria over 590 s. CONCLUSIONS: The quantitative analysis demonstrated a significant decrease of S. mutans CFU at 90 s PB exposure and only after 300 s of Se application. Protect Bond showed an earlier antibacterial effect than Se Bond.

Analysis by confocal laser scanning microscopy of the MDPB bactericidal effect on S. mutans biofilm CLSM analysis of MDPB bactericidal effect on biofilm

Since bacteria remain in the dentin following caries removal, restorative materials with antibacterial properties are desirable to help maintaining the residual microorganisms inactive. The adhesive system Clearfil Protect Bond (PB) contains the antibacterial monomer 12-methacryloyloxydodecylpyridinium bromide (MDPB) in its primer, which has shown antimicrobial activity. However, its bactericidal effect against biofilm on the dentin has been little investigated. Objective: The aim of this study was to analyze by confocal laser scanning microscopy (CLSM) and viable bacteria counting (CFU) the MDPB bactericidal effect against S. mutans biofilm on the dentin surface. Material and methods: Bovine dentin surfaces were obtained and subjected to S. mutans biofilm formation in BHI broth supplemented with 1% (w/v) sucrose for 18 h. Samples were divided into three groups, according to the primer application (n=3): Clearfil Protect Bond (PB), Clearfil SE Bond, which does not contain MDPB, (SE) and saline (control group). After the biofilm formation, Live/ Dead stain was applied directly to the surface of each sample. Next, 10 µL of each primer were applied on the samples during 590 s for the real-time CLSM analysis. The experiment was conducted in triplicate. The primers and saline were also applied on the other dentin samples during 20, 90, 300 and 590 s (n=9 for each group and period evaluated) and the CFU were assessed by colonies counting. Results: The results of the CLSM showed that with the Se application, although non-viable bacteria were detected at 20 s, there was no increase in their count during 590 s. In contrast, after the PB application there was a gradual increase of non-viable bacteria over 590 s. Conclusions: The quantitative analysis demonstrated a significant decrease of S. mutans CFU at 90 s PB exposure and only after 300 s of Se application. Protect Bond showed an earlier antibacterial effect than Se Bond.

Marginal adaptation of indirect restorations using different resin coating protocols

This study evaluated the influence of material combinations used in the resin coating technique (RCT) on the marginal adaptation of indirect restorations with gingival margins in enamel (EM) and cement (CM). Eighty third-molars were used. Two cavities were prepared in each tooth. The cavities were distributed into 16 groups. Cavities with EM were filled with the following material combinations: G1: Single-Bond 2 (Sb2), G2: Sb2 + Bond/Scotchbond-Multipurpose (Sb2B), G3: Sb2 + Filtek-Flow Z350 (Sb2Fl), G4: Scotchbond-Multipurpose (SBMP), G5: Clearfil-S3 (CS3), G6: CS3 + Bond/Clearfil-SE Bond (CSE3B), G7: CS3 + Protect Liner F (CS3PL) and G8: Clearfil SE Bond + Protect Liner F (CSEBPL). The same combinations were applied to the cavities in CM: G9, G10, G11, G12, G13, G14, G15, G16, respectively. The fillings were performed with the Sinfony-System (3M/ESPE). After 24 h, the teeth were submitted to thermocycling (2,000 cycles, 5° to 55°C) and load-cycling (50,000 cycles, 50 N). Next, the Caries-Detector (Kuraray) was applied to the restoration margins. Images from the proximal margin were evaluated using the Image-Tool 3.0 software. The results were submitted to ANOVA and Tukey's test (α=0.05). The mean values (%) for the groups were: EM: G1=46.68, G2=15.53, G3=19.83, G4=27.53; G5=59.49, G6=25.13, G7=34.37, G8=15.20; CM: G9=38.38, G10=23.25, G11=26.97, G12=25.85, G13=37.81, G14=30.62, G15=29.17, G16=20.31. The highest percentages of marginal gap on EM or CM were found in the groups that did not use a liner. It can be concluded that the most appropriate RCT combinations were the groups that used a liner.

Este estudo avaliou a influência de diferentes combinações de materiais usados na técnica de selamento dentinário (TSD) sobre a adaptação marginal de restaurações indiretas, cujas margens gengivais localizam-se em esmalte (ME) ou cemento (MC). Oitenta terceiros molares foram selecionados e duas cavidades foram preparadas em cada dente, as quais foram distribuídas em 16 grupos. As cavidades com margem em esmalte foram forradas pelas seguintes combinações de materiais: G1: Single-Bond2 (Sb2), G2: Sb2 + Bond/Scotchbond-Multipurpose (Sb2B), G3: Sb2 + Filtek-Flow Z350 (Sb2Fl), G4: Scotchbond-Multipurpose (SBMP), G5: Clearfil-S3 (CS3), G6: CS3 + Bond/Clearfil-SE Bond (CSE3B), G7: CS3 + Protect Liner F (CS3PL), G8: Clearfil SE Bond + Protect Liner F (CSEBPL). As mesmas combinações foram aplicadas às cavidades com margens em cemento: G9, G10, G11, G12, G13, G14, G15, G16, respectivamente. As restaurações foram confeccionadas usando o sistema Sinfony (3M/ESPE). Após 24 h, os dentes restaurados foram submetidos à ciclagem térmica (2.000 ciclos - 5° a 55° C) e mecânica (50.000 ciclos, 50 N). Em seguida, Carie-Detector (Kuraray) foi aplicado sobre as margens das restaurações. As imagens obtidas da margem proximal foram avaliadas pelo software Image-Tool 3.0. Os resultados foram submetidos aos testes estatísticos ANOVA e Tukey (p<0,05). As médias (%) observadas para os grupos foram: ME: G1=46,68, G2=15,53, G3=19,83, G4=27,53; G5=59,49, G6=25,13, G7=34,37 e G8=15,20; MC: G9=38,38, G10=23,25, G11=26,97, G12=25,85, G13=37,81, G14=30,62, G15=29,17, G16=20,31. Os maiores valores de desadaptação marginal encontrados em ME e MC foram encontrados nos grupos que não utilizaram um "liner". Desta forma, pôde-se concluir que a combinação mais apropriada para a TSD é aquela que faz uso do "liner".

Effect of gamma irradiation on fluoride release and antibacterial activity of resin dental materials.

This study evaluated the effect of gamma irradiation on fluoride release and antibacterial activity of FluroShield (FS) and Clearfil Protect Bond (CPB). Four groups were formed: G1-FS + gamma; G2-FS without gamma; G3-CPB + gamma; G4-CPB without gamma. For fluoride release analysis, 12 disks of each material were prepared and covered with nail polish, except for one side (50.4 mm(2) area). G1 and G3 were sterilized with a 14.5 KGy dose at 27 degrees C for 24 h, while G2 and G4 (controls) were not sterilized and were maintained under the same time and temperature conditions. Fluoride release measurements were made in duplicate (n=6) by an ion specific electrode. The antibacterial activity of the CPB and FS against Streptococcus mutans after gamma sterilization was evaluated by the agar-disc diffusion method. The diameter of the zones of microbial growth inhibition was recorded after 48 h. Data were analyzed statistically by ANOVA and Tukey's test (alpha=5%). Gamma sterilization decreased the fluoride release of FS by approximately 50%, while CPB was not affected. There was no statistically significant difference (p>0.05) in the antibacterial effect of CPB between gamma and non-gamma sterilization groups. FS presented no antibacterial activity. Gamma irradiation decreased the fluoride release of FS, but did not affect the antibacterial activity of the studied materials.

Effect of gamma irradiation on fluoride release and antibacterial activity of resin dental materials

This study evaluated the effect of gamma irradiation on fluoride release and antibacterial activity of FluroShield (FS) and Clearfil Protect Bond (CPB). Four groups were formed: G1-FS + gamma; G2-FS without gamma; G3-CPB + gamma; G4-CPB without gamma. For fluoride release analysis, 12 disks of each material were prepared and covered with nail polish, except for one side (50.4 mm² area). G1 and G3 were sterilized with a 14.5 KGy dose at 27ºC for 24 h, while G2 and G4 (controls) were not sterilized and were maintained under the same time and temperature conditions. Fluoride release measurements were made in duplicate (n=6) by an ion specific electrode. The antibacterial activity of the CPB and FS against Streptococcus mutans after gamma sterilization was evaluated by the agar-disc diffusion method. The diameter of the zones of microbial growth inhibition was recorded after 48 h. Data were analyzed statistically by ANOVA and Tukey's test (α=5 percent). Gamma sterilization decreased the fluoride release of FS by approximately 50 percent, while CPB was not affected. There was no statistically significant difference (p>0.05) in the antibacterial effect of CPB between gamma and non-gamma sterilization groups. FS presented no antibacterial activity. Gamma irradiation decreased the fluoride release of FS, but did not affect the antibacterial activity of the studied materials.

Este estudo avaliou o efeito da esterilização com raios-gama na liberação de flúor e atividade antibacteriana de materiais resinosos, Fluroshield (FS) e Clearfil Protect Bond (CPB). Quatro grupos foram formados: G1-FS e gama; G2-FS sem gama; G3-CPB e gama; G4-CPB sem gama. Doze discos de cada material foram preparados para análise de liberação de flúor, os quais foram cobertos com esmalte de unha, exceto em um lado com 50,4 mm² de área. G1 e G3 foram esterilizados com dose de 14,5 KGy por 24 h/27ºC, enquanto G2 e G4 (controles) não foram esterilizados e foram mantidos sob as mesmas condições de tempo e temperatura. As leituras de liberação de flúor foram feitas em duplicata (n=6) por um eletrodo específico. A atividade antibacteriana foi avaliada pelo teste de difusão em agar. Os halos de inibição foram medidos após 48 h. Os dados foram analisados pelos testes ANOVA e Tukey (α=5 por cento). A esterilização gama diminuiu a liberação de flúor de FS em cerca de 50 por cento, enquanto CPB não foi afetado. Não houve diferença estatisticamente significante entre os grupos esterilizados e controle no efeito antibacteriano do CPB. FS não apresentou atividade antibacteriana. A esterilização gama diminuiu a liberação de flúor de FS, mas não afetou a atividade antibacteriana dos materiais estudados.

Confocal laser scanning microscopic analysis of the depth of dentin caries-like lesions in primary and permanent teeth

This study analyzed comparatively, by confocal laser scanning microscopy (CLSM), the depth of caries-like lesions produced by biological and chemical artificial models in permanent and primary dentin. Six primary molars and six premolars were used. The occlusal enamel was removed and a nail polish layer was applied on the specimens, except for a 4 x 2 mm area on dentin surface. Half of specimens were immersed in acid gel for 14 days (chemical model) and the other half was immersed in BHI broth with S. mutans for 14 days (biological model). After development of artificial caries, the crowns were longitudinally sectioned on the center of the carious lesion. Three measurements of carious dentin depth were made in each specimen by CLSM. Measurements depths were compared between the caries models and between tooth types by one-way ANOVA and Tukey test (a=5 percent). For permanent teeth, the biological model showed significantly higher (p<0.05) caries depth values than the chemical model. For primary teeth, no statistically significant difference (p>0.05) was found between the caries models. The artificial caries model influenced caries depth only in permanent teeth. There was no difference in carious dentin depth between permanent and primary teeth, regardless of the artificial caries model.

O objetivo deste estudo foi comparar a profundidade de cárie produzida em dentina permanente e decídua por modelos biológico e químico de produção artificial de cárie utilizando o microscópio confocal de varredura a laser (CLSM, na sigla em Inglês). Seis molares decíduos e seis pré-molares foram usados. O esmalte oclusal foi removido expondo a dentina subjacente. A seguir, um verniz de unha foi aplicado em toda a amostra, exceto em uma área de 4 x 2 mm na superfície dentinária. Metade das amostras foi imersa em gel ácido por 14 dias (modelo químico) e a outra metade imersa em BHI com S. mutans por 14 dias (modelo biológico). Após o desenvolvimento da cárie artificial, as coroas foram seccionadas longitudinalmente no centro da lesão de cárie. Três medidas da profundidade de cárie produzida foram realizadas ao longo de cada espécime e analisadas em CLSM. As medidas da profundidade de cárie entre os modelos e entre os tipos de dentes foram analisadas pelo teste de ANOVA a um critério e teste de Tukey (p<0,05). Para os dentes permanentes, o modelo biológico mostrou maiores valores de profundidade de cárie quando comparado ao modelo químico. Entretanto, para os dentes decíduos não houve diferença estatisticamente significante na profundidade de cárie entre os modelos. Desta forma, o modelo de produção de cárie artificial influenciou a profundidade de cárie apenas para os dentes permanentes, não existindo diferença na profundidade de cárie entre dentes decíduos e permanentes, independente do modelo de cárie utilizado.