Papain gel containing methylene blue for simultaneous caries removal and antimicrobial photoinactivation against Streptococcus mutans biofilms

This study intended to evaluate the effects of a papain-gel with a red-light absorbing pigment (methylene blue – MB) to mediate photodynamic therapy (PDT) against Streptococcus mutans biofilms. The PapaMBlue was compared with free MB to generate reactive oxygen species using fluorescence probes (SOSG and HPF). PDT (660-nm light) was carried out against S. mutans biofilms grown on either plastic dishes or on collagen membrane and assayed by CFU, live-dead staining using confocal microscopy, transmission electron microscopy and H&E staining for collagen films. Cytotoxicity and subcellular localization was studied in human fibroblasts. Sponges of bioabsorbable type I collagen membrane were exposed to papain based gel, irradiated with laser and analyzed about their integrity by ATR-FTIR. The PapaMBlue produced higher amounts of singlet oxygen and hydroxyl radicals than free MB, possibly due to better disaggregation of the dye in solution. The PapaMBlue antimicrobial effects on biofilms proved to be capable of reducing the S. mutans. Both MTT and PrestoBlue assays showed higher cell viability and metabolism scores in fibroblasts treated with PapaMBlue and MB, possibly due to stimulation of mitochondrial activity and that collagen triple helix is unaffected. The PapaMBlue is equally effective as MB in destroying S. mutans biofilms growing on plastic or collagen without affecting fibroblasts.National Institutes of Health (U.S.) (Grant R01AI050875

Er,Cr:YSGG laser irradiation influence on enamel and dentin bond strength of adhesive systems

Este trabalho teve o objetivo de avaliar a resistência adesiva de sistemas adesivos baseados no condicionamento prévio com ácido fosfórico (etch-and-rinse) e auto-condicionantes (de um e dois passos) ao substrato dental, submetido a diferentes condições de irradiação com laser de Er,Cr:YSGG. Amostras de esmalte e dentina foram obtidas do terço médio da face vestibular de 216 incisivos bovinos, divididos aleatoriamente em 18 grupos (n=12), de acordo com o substrato, (esmalte e dentina), sistema adesivo (etch-and-rinse, auto-condicionante de um passo e auto-condicionante de dois passos) e condições de irradiação (sem irradiação ? controle, irradiado com refrigeração ar-água e irradiado sem refrigeração ar-água). Em seguida foi realizada inclusão e polimento, sob refrigeração, com lixas de carbeto de silício de granulação decrescente 180, 320, 400 e 600 (por um minuto para padronização da camada de esfregaço). Para os grupos irradiados com laser de Er,Cr:YSGG (?=2,79 ?m, taxa de repetição de 20 Hz), com densidade de energia de 2,8 J/cm2, 12,5 mJ por pulso, potência média de 0,25W, com a ponta S75 de diâmetro de 750 ?m a 1 mm da superfície do substrato. A condição de refrigeração da irradiação foi realizada com 11 mL/min. Os corpos-de-prova foram confeccionados com a resina composta Filtek Z250, em forma de cone invertido para realização do teste de resistência adesiva. Após 24 h de armazenagem em água destilada a 37ºC, fez-se o ensaio de tração a 0,5 mm/min. Os dados foram comparados estatisticamente pelo método de ANOVA com significância de 5%. Os resultados obtidos mostraram que para o tecido esmalte foi observada diferença estatisticamente significante para os fatores adesivo e condições de irradiação empregadas, sem significância para a interação entre os fatores. A resistência de união do sistema adesivo auto-condicionante de dois passos e do sistema adesivo etch-and-rinse apresentam desempenho semelhante, sendo superiores ao sistema adesivo auto-condicionante de passo único. Ainda para esmalte, o efeito das condições de irradiação mostrou que os grupos sem irradiação com laser de Er,Cr:YSGG apresentaram maior resistência adesiva do que os grupos irradiados com refrigeração e sem refrigeração. Para o substrato dentina, houve diferença estatisticamente significante para os fatores de variação principais: sistemas adesivos, condição de irradiação com laser de Er,Cr:YSGG e para a interação adesivo X condição de irradiação. Quanto à comparação entre os sistemas adesivos testados na condição sem irradiação, os sistemas adesivos auto-condicionantes, de dois passos e o de um passo, não foram estatisticamente diferentes entre si, porém ambos forneceram resultados de resistência adesiva estatisticamente menores do que o sistema adesivo etch-and-rinse. Para as condições de irradiação com laser de Er,Cr:YSGG, com refrigeração e sem refrigeração, obteve-se valores de resistência adesiva maiores para o sistema auto-condicionante de dois passos do que para os demais sistemas. Deste modo, conclui-se que a irradiação com laser de Er,Cr:YSGG reduz a adesão ao esmalte independentemente do sistema adesivo utilizado e da condição de irradiação. Adicionalmente, observa-se que a irradiação de dentina com laser de Er,Cr:YSGG somente não altera a resistência adesiva do sistema auto-condicionante de dois passos, sendo deletéria aos demais adesivos testados.The aim of this study was to assess the bond strength to dental substrate of adhesives based on previous etching with phosphoric acid (etch-and-rinse) and self-etching (one- and two-step) under different conditions of Er,Cr:YSGG laser irradiation. Enamel and dentin samples were obtained from the middle third of the buccal face of 216 bovine incisors, randomly divided in 18 groups (n=12), according to the substrate (enamel and dentin), adhesive system (etch-and-rinse, one-step self-etching and two-step self-etching) and irradiation condition (without irradiation ? control, irradiated with cooling, and irradiated without cooling). Next, the samples were included and polished, under cooling, with silicone carbide abrasive papers of decreasing grit #180, 320, 400 and 600 (for one minute to standardize the smear layer). For the irradiated groups with Er,Cr:YSGG laser, an energy density of 2.8 J/cm2, 12.5 mJ per pulse, mean power of 0.25W was selected. During laser irradiation, cooling was performed with 11 mL/min. Inverted cone-shaped test specimens were built-up with resin composite Filtek Z250, for performing the bond strength test. After 24 h storage in distilled water at 37ºC, the tensile test was performed at 0.5 mm/min. The results obtained show, for enamel, statistically significant difference for the factors adhesive and irradiation conditions used, without significance for the interaction among the factors. The bond strength of the 2-step self-etching and the etch-and-rinse adhesive system presented similar performance, being superior to that of one-step self-etching system. For enamel, the effect of the irradiation conditions showed that the groups without Er,Cr:YSGG laser irradiation presented higher bond strength than the groups irradiated with and without cooling. For the dentin substrate there was statistically significant difference for the main variation factors: adhesive systems, condition of Er,Cr:YSGG laser irradiation and for the interaction adhesive X irradiation condition. As regards the comparison among the adhesive systems tested under the condition without irradiation, the one-step and two-step self-etching adhesive systems did not differ statistically between them, but both provided statistically lower bond strength results than the etch-and-rinse adhesive system. For the condition of Er,Cr:YSGG laser irradiation with and without cooling, higher bond strength values were obtained for the two-step self-etching system than for the other systems. Therefore, it was concluded that Er,Cr:YSGG laser irradiation reduces the bond strength to enamel irrespective of the adhesive system used and the irradiation condition. Furthermore, it was observed that dentin irradiated with Er,Cr:YSGG laser did not alter the bond strength of the two-step adhesive system only, and it was deleterious to the other adhesives tested

Longitudinal evaluation of the effect of saliva contamination during the bonding protocol with a self-etch adhesive system

Aim: To evaluate the influence of saliva contamination on the short- and long-term bond strength of a self-etch adhesive system. Methods: One hundred and twelve non-carious human molars were randomly divided according to: substrate (enamel/dentin); presence of saliva [none (control-C), before primer (BP), after primer (AP) and after bonding agent (AB)]; treatment of the contamination [none (1), rinsing + drying (2), drying (3) and primer re-application (4)] and specimen storage (24 h or 6 months). A self-etch adhesive system was applied to the dental surfaces followed by incremental insertions of composite resin. After storage in water at 37 o C, the specimens were perpendicularly cut into beams for microtensile bond strength testing. Data in MPa were compared by ANOVA followed by Tukey′s test (p< 0.05). Micrographs were obtained by low vacuum scanning electron microscopy. Results: Control groups (G1 and G8) presented higher bond strength than all other groups. The factors presence of saliva, treatments of the contaminant and specimen storage showed no statistically significant results for the two dental substrates. Contaminants could be detected by LV-SEM. Six-month storage did not affect bond strength. Conclusions: The presence of saliva during the application of the self-etch system was deleterious to the bond to enamel and dentin, irrespective of the operative step in which the contamination occurred

Longitudinal evaluation of the effect of saliva contamination during the bonding protocol with a self-etch adhesive system

evaluate the influence of saliva contamination on the short- and long-term bond strength of a self-etch adhesive system. Methods: One hundred and twelve non-carious human molars were randomly divided according to: substrate (enamel/dentin); presence of saliva [none (control-C), before primer (BP), after primer (AP) and after bonding agent (AB)]; treatment of the contamination [none (1), rinsing + drying (2), drying (3) and primer re-application (4)] and specimen storage (24 h or 6 months). A self-etch adhesive system was applied to the dental surfaces followed by incremental insertions of composite resin. After storage in water at 37oC, the specimens were perpendicularly cut into beams for microtensile bond strength testing. Data in MPa were compared by ANOVA followed by Tukeys test (p< 0.05). Micrographs were obtained by low vacuum scanning electron microscopy. Results: Control groups (G1 and G8) presented higher bond strength than all other groups. The factors presence of saliva, treatments of the contaminant and specimen storage showed no statistically significant results for the two dental substrates. Contaminants could be detected by LV-SEM. Six-month storage did not affect bond strength. Conclusions: The presence of saliva during the application of the self-etch system was deleterious to the bond to enamel and dentin, irrespective of the operative step in which the contamination occurred

Adhesion after erbium, chromium:yttrium-scandium-gallium-garnet laser application at three different irradiation conditions

The aim of this study was to investigate whether distinct cooling of low fluence erbium, chromium:yttrium-scandium-gallium-garnet (Er,Cr:YSGG) laser irradiation would influence adhesion. Main factors tested were: substrates (two), irradiation conditions (three), and adhesives (three). A 750 mu m diameter tip was used, for 50 s, 1 mm from the surface, with a 0.25 W power output, 20 Hz, energy density of 2.8 J/cm(2) with energy per pulse of 12.5 mJ. When applied, water delivery rate was 11 ml/min. The analysis of variance (ANOVA) showed that laser conditioning significantly decreased the bond strength of all adhesive systems applied on enamel. On dentin, laser conditioning significantly reduced bond strength of etch-and-rinse and one-step self-etch systems; however, laser irradiation under water cooling did not alter bonding of two-step self-etching. It may be concluded that the irradiation with Er,Cr:YSGG laser at 2.8 J/cm(2) with water coolant was responsible for a better adhesion to dentin, while enamel irradiation reduced bond strength, irrespective of cooling conditions.Funda ao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP)[2003/12182-4]Funda ao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP)[2004/02229-6

Relationship Between Surface Topography and Energy Density Distribution of Er,Cr:YSGG Beam on Irradiated Dentin: An Atomic Force Microscopy Study

Objective: The aim of this study was to assess by atomic force microscopy (AFM) the effect of Er,Cr:YSGG laser application on the surface microtopography of radicular dentin. Background: Lasers have been used for various purposes in dentistry, where they are clinically effective when used in an appropriate manner. The Er, Cr: YSGG laser can be used for caries prevention when settings are below the ablation threshold. Materials and Methods: Four specimens of bovine dentin were irradiated using an Er, Cr:YSGG laser (lambda = 2.78 mu m), at a repetition rate of 20 Hz, with a 750-mu m-diameter sapphire tip and energy density of 2.8 J/cm(2) (12.5 mJ/pulse). After irradiation, surface topography was analyzed by AFM using a Si probe in tapping mode. Quantitative and qualitative information concerning the arithmetic average roughness (Ra) and power spectral density analyses were obtained from central, intermediate, and peripheral areas of laser pulses and compared with data from nonirradiated samples. Results: Dentin Ra for different areas were as follows: central, 261.26 (+/- 21.65) nm; intermediate, 83.48 (+/- 6.34) nm; peripheral, 45.8 (+/- 13.47) nm; and nonirradiated, 35.18 (+/- 2.9) nm. The central region of laser pulses presented higher ablation of intertubular dentin, with about 340-760 nm height, while intermediate, peripheral, and nonirradiated regions presented no difference in height of peritubular and interperitubular dentin. Conclusion: According to these results, we can assume that even when used at a low-energy density parameter, Er, Cr: YSGG laser can significantly alter the microtopography of radicular dentin, which is an important characteristic to be considered when laser is used for clinical applications.Fundacao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP)[1995/5651-0]Fundacao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP)[2006/05684-1]Fundacao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP)[2006/06746-0]Conselho Nacional de Desenvolvimento Cientifico e Tecnologico (CNPq)[473723/2007-7]Conselho Nacional de Desenvolvimento Cientifico e Tecnologico (CNPq)[143395/2009-2