Performance of self-etching adhesives on caries-affected primary dentin treated with glutaraldehyde or silver diamine fluoride.

OBJECTIVES: to evaluate the quality and stability of adhesive interfaces established by self-etching adhesives on caries-affected primary dentin (CAD) treated with glutaraldehyde (GA) or silver diamine fluoride (SDF). METHODS: 42 primary molars were exposed to a microbiological caries-inducing protocol and divided into 6 groups according to the adhesive system (Clearfil SE - CL or FL Bond II - FL) and pretreatment (water, GA or SDF) applied on CAD. One tooth from each group was analyzed for surface modification using infrared spectroscopy. Crowns were restored with resin composite (n = 36) and cut into beams and slices. The beams were subjected to microtensile testing, Raman spectroscopy and SEM after 24 h and 6 months of storage. The slices were analyzed using Micro-Raman spectroscopy to determine the diffusion zone thickness (DZ) in each period. Data were analyzed by ANOVA and Tukey or Kruskal-Wallis and Dunn tests (α = 0.05%). RESULTS: SDF reduced the immediate bond strength for both adhesives. The control groups showed a decrease in BS after 6 months in artificial saliva. GA increased immediate DZ for FL, while SDF had the opposite effect on CL. GA decreased the DZ for FL at 6 months. There was a predominance of adhesive failures with areas of cohesive dentin fractures within control groups. SIGNIFICANCE: Modifications caused by dentin surface treatments may directly affect the performance of adhesive systems and the quality and stability of adhesive restorations.

Inhibitory activity of S-PRG filler on collagen-bound MMPs and dentin matrix degradation.

OBJECTIVES: To evaluate the inhibitory activity of an ion-releasing filler (S-PRG) eluate on dentin collagen-bound metalloproteinases (MMPs) and dentin matrix degradation. METHODS: Dentin beams (5 × 2 × 0.5 mm) from human molars were completely demineralized to produce dentin matrix specimens. The dry mass was measured, and a colorimetric assay (Sensolyte) determined the initial total MMP activity to allocate the beams into four treatment groups (n = 10/group): 1) water for 1 min (negative control); 2) 2% chlorhexidine digluconate (CHX - inhibitor control) for 1 min; 3) S-PRG eluate for 1 min; 4) S-PRG eluate for 30 min. After the treatments, the total MMP activity was reassessed. The specimens were stored in simulated body fluid (SBF) at 37 °C for up to 21 days. The dry mass was reassessed weekly. On day 7, the dentin matrix degradation was analyzed for the presence of collagen fragments (CF; Sirius Red) and hydroxyproline (Hyp) in the SBF. Statistical analyses were performed with ANOVA/Tukey, paired t-tests, and RM-ANOVA/Sidak (α = 5%). RESULTS: S-PRG eluate exposure for 1 and 30 min reduced (p < 0.0001) MMP activity. S-PRG exposure for 30 min presented MMP activity inhibition equivalent to CHX (p = 0.061). S-PRG and CHX decreased CF (p ≤ 0.007) and Hyp (p < 0.046) release. After 21 days of storage, S-PRG-treated beams, regardless of exposure time, presented a reduced (p ≤ 0.017) mass loss, intermediate between CHX and control. CONCLUSION: Treating demineralized dentin with S-PRG eluate for 1 or 30 min reduced matrix-bound MMP activity and dentin matrix degradation for up to 21 days. CLINICAL SIGNIFICANCE: S-PRG filler may hinder the progression of dentin carious/erosive lesions and enhance the stabilization of dentin bonding interfaces.

Proteolytic activity and degradation of bovine versus human dentin matrices.

OBJECTIVE: Non-human teeth have been commonly used in research as replacements for human teeth, and potential dissimilarities between the dental tissues should be considered when interpreting the outcomes. To compare the proteolytic activity and degradation rate of bovine and human dentin matrices. METHODOLOGY: Dentin beam specimens were obtained from human molars (n=30) and bovine incisors (n=30). The beams were weighed hydrated and after complete dehydration to obtain the mineralized wet and dry masses. Then, the beams were demineralized in 10 wt% phosphoric acid. Next, 15 beams from each substrate were randomly selected and again dehydrated and weighed to obtain the initial demineralized dry mass (DM). Then, the beams were stored in saliva-like buffer solution (SLBS) for 7, 14 and 21 days. SLBS was used to evaluate hydroxyproline (HYP) release after each storage period. The remaining beams of each substrate (n=15) were tested for initial MMP activity using a colorimetric assay and then also stored in SLBS. DM and MMP activity were reassessed after 7, 14 and 21 days of incubation. The data were subjected to two-way ANOVA tests with repeated measures complemented by Bonferroni's tests. Unpaired two-tailed t-tests were also used (p<0.05). RESULTS: Similar water and inorganic fractions were found in human and bovine dentin, while human dentin had a higher protein content. The most intense proteolytic activity and matrix deterioration occurred short after dentin was demineralized. Both substrates exhibited a sharp reduction in MMP activity after seven days of incubation. Although human dentin had higher MMP activity levels, greater HYP release and DM loss after seven days than bovine dentin, after 14 and 21 days, the outcomes were not statistically different. CONCLUSION: Bovine dentin is a suitable substrate for long-term studies involving the degradation of dentin matrices.

Response of pulp cells to resin infiltration of enamel white spot-like lesions.

OBJECTIVES: To investigate the trans-enamel and trans-dentinal biological effects of treating enamel white spot-like lesions (EWSLs) with resin infiltration components (RICs) on odontoblast-like cells (MDPC-23) and human dental pulp cells (HDPCs). METHODS: EWSLs were induced in 60 enamel/dentin discs (4.0 ± 0.2 mm thick) using S. mutans. The discs were adapted into artificial pulp chambers and MDPC-23 were seeded on the dentin surface. The components of a resin infiltration system (Icon) were applied individually or in combination on the enamel surface as following (n = 10/treatment): Etch, Infiltrant, Etch+Infiltrant, or Etch+Dry+Infiltrant. The application of water or hydrogen peroxide served as negative and positive controls, respectively. After 72 h, MDPC-23 viability was evaluated. The extracts were exposed for 72 h to pre-cultured MDPC-23 and HDPCs in 96-well plates to evaluate cell viability, alkaline phosphatase activity (ALP), mineralized nodule formation (MN), and the expression of inflammatory cytokines (ICs) and mineralization-related genes (MRs). Data were analyzed by ANOVA complemented with Tukey or Games-Howell post-hocs (α = 5%). RESULTS: Cell viability, ALP activity, and MN formation were significantly reduced in response to the RICs, presenting intermediate values compared to positive and negative controls. Likewise, ICs were upregulated, whereas MRs were downregulated. Among the RICs, the Etch component caused the most notorious detrimental effects. SIGNIFICANCE: Resin infiltration of EWSLs negatively affected the metabolism of pulp cells in vitro. Therefore, even though resin infiltration is a micro-invasive therapy for non-cavitated caries in enamel, it should be closely followed up seen that components may diffuse and unbalance pulp homeostasis.

Proteolytic activity and degradation of bovine versus human dentin matrices

Abstract Non-human teeth have been commonly used in research as replacements for human teeth, and potential dissimilarities between the dental tissues should be considered when interpreting the outcomes. Objective: To compare the proteolytic activity and degradation rate of bovine and human dentin matrices. Methodology: Dentin beam specimens were obtained from human molars (n=30) and bovine incisors (n=30). The beams were weighed hydrated and after complete dehydration to obtain the mineralized wet and dry masses. Then, the beams were demineralized in 10 wt% phosphoric acid. Next, 15 beams from each substrate were randomly selected and again dehydrated and weighed to obtain the initial demineralized dry mass (DM). Then, the beams were stored in saliva-like buffer solution (SLBS) for 7, 14 and 21 days. SLBS was used to evaluate hydroxyproline (HYP) release after each storage period. The remaining beams of each substrate (n=15) were tested for initial MMP activity using a colorimetric assay and then also stored in SLBS. DM and MMP activity were reassessed after 7, 14 and 21 days of incubation. The data were subjected to two-way ANOVA tests with repeated measures complemented by Bonferroni's tests. Unpaired two-tailed t-tests were also used (p<0.05). Results: Similar water and inorganic fractions were found in human and bovine dentin, while human dentin had a higher protein content. The most intense proteolytic activity and matrix deterioration occurred short after dentin was demineralized. Both substrates exhibited a sharp reduction in MMP activity after seven days of incubation. Although human dentin had higher MMP activity levels, greater HYP release and DM loss after seven days than bovine dentin, after 14 and 21 days, the outcomes were not statistically different. Conclusion: Bovine dentin is a suitable substrate for long-term studies involving the degradation of dentin matrices.

COVID-19 pandemic and pediatric dentistry: Fear, eating habits and parent's oral health perceptions.

BACKGROUND: Severe measures have been implemented around the world to reduce COVID-19 spread with a significant impact on family dynamics. AIM: To assess the impact of the pandemic on fear, dietary choices and oral health perceptions of parents. DESIGN: questionnaire containing 19 questions was remotely applied to 1003 parents of children aged 0-12 years. The questions addressed topics regarding changes in daily routine, dietary habits, fear level, oral health, and variation of income during the pandemic. Data analysis included the description of the relative and absolute frequencies of the variables. Association tests were performed using Fisher's exact and Kruskal-Wallis tests. RESULTS: 73% of respondents reported income loss. Five hundred sixty-eight people denied seeking medical or dental care. 61.5% of respondents revealed changes in the dietary pattern; most of them mentioned an increase in food intake. Most parents (66.6%) would only seek urgent dental care. There was an association between parents' willingness to take their children to dental appointments with the fear level (p < 0.001). CONCLUSIONS: Most families have experienced changes in daily routine and eating habits during the pandemic. Parents fear COVID-19 and it impacts their behavior regarding seeking dental care for their children.

Proteolytic activity, degradation, and dissolution of primary and permanent teeth.

BACKGROUND: Primary and permanent teeth composition may influence dissolution and degradation rates. AIM: To compare the dissolution and degradation of primary and permanent teeth. DESIGN: Enamel and dentin powders were obtained from primary molars and premolars and incubated within different pH buffers. Calcium and inorganic phosphate release was quantified in the buffers by atomic absorption and light spectrophotometry. A colorimetric assay was used to assess the MMP activity of primary dentin (PrD) and permanent dentin (PeD). Collagen degradation was assessed by dry mass loss, change in elastic modulus (E), and ICTP and CTX release. Data were submitted to ANOVA and Tukey's tests (α = 0.05). RESULTS: Similar dissolution was found between PrD and PeD after 256 hours. At pH 4.5, enamel released more minerals than dentin whereas at pH 5.5 the inverse result was observed. MMP activity was similar for both substrates. PrD showed higher dry mass loss after 1 week. In general, greater reduction in E was recorded for PrD. Higher quantities of ICTP and CTX were released from PrD after 1 week. CONCLUSIONS: Primary and permanent teeth presented similar demineralization rates. Collagen degradation, however, was faster and more substantial for PrD.

Effect of Er:YAG laser irradiation and chitosan biomodification on the stability of resin/demineralized bovine dentin bond.

OBJECTIVE: The purpose of this study was to evaluate the effect of Er:YAG laser for selective removal of carious lesion, followed by biomodification with chitosan on the microtensile bond strength (µTBS), adhesive interface, dry mass loss and hydroxyproline release (HYP). METHODS: Artificial lesions were created in 104 bovine dentin blocks. Blocks were divided according to caries removal method: bur or Er:YAG laser. Seventy-six blocks were acid etched and subdivided according to dentin biomodification: no chitosan and 2.5% chitosan. Composite resin restorations were performed. Blocks were sectioned into beams and stored in water. After 24 h, 6 and 12 months, beams were submitted to µTBS test (n = 10) and analysis of adhesive interface by SEM (n = 3). The other 28 blocks were sectioned into beams and initial dry mass (DM) was determined (n = 7). Beams were stored and after 7 days, DM was redetermined. HYP release (n = 7) was evaluated by ELISA. Data were analyzed by ANOVA and Bonferroni's tests (α = 0.05). RESULTS: After 24 h, the highest µTBS was found for bur (p < 0.001). After 6 months, methods were similar (p = 0.432). After 12 months, laser-irradiated dentin showed the highest µTBS values (p = 0.025). Chitosan promoted higher µTBS values after 6 (p = 0.011) and 12 months (p < 0.001) preserving adhesive interface. Dry mass loss and HYP release were not influenced (p > 0.05) by caries removal method or by dentin biomodification. CONCLUSION: The bond strength to demineralized dentin reduced over 50% in all groups after water storage. From 6 months of water storage, Er:YAG laser irradiation and biomodification with chitosan maintained the stability of the resin-dentin bonds, but did not influence dry mass loss and HYP release.

Redução da atividade proteolítica da dentina após curtos períodos de aplicação de proantocianidina / Reduction of the dentin proteolytic activity after short periods of application of proanthocyanidin

Introdução: Agentes promotores de ligações cruzadas têm sido investigados como inibidores da atividade enzimática da dentina, o que favoreceria a longevidade das restaurações adesivas. Objetivo: Avaliar o efeito do tratamento da dentina com proantocianidina (PA), em curtos períodos de tempo, na inibição da atividade de MMPs in situ. Material e método: Quarenta espécimes de dentina (1×1×6 mm) foram obtidos de molares hígidos e divididos em quatro grupos (n=10). Os espécimes foram condicionados com ácido fosfórico por 15 s, seguido de lavagem em água deionizada. A dentina condicionada foi tratada com: água, 5% PA por 5 s, 15 s ou 30 s. A atividade de MMP foi analisada colorimetricamente (SensoLyte®) e os dados de absorbância (412 nm) foram submetidos aos testes de ANOVA e Tukey (alfa =0,05). Resultado: Todos os períodos de tratamento foram capazes de reduzir a atividade de MMPs, sendo que os melhores resultados foram observados para a dentina tratada com PA por 15 s (63,1% redução) e 30 s (70,2%). O tratamento por 5 s foi capaz de inibir 39,9% das MMPs. Conclusão: A aplicação de PA sobre a dentina condicionada foi capaz de reduzir a atividade de MMPs mesmo em períodos de tempo extremamente curtos, como 5 s. No entanto, melhores resultados foram obtidos com os maiores períodos de tratamento.

Introduction: Collagen cross-linkers have been investigated as inhibitors of the enzymatic activity of dentin, therefore improving the longevity of adhesive restorations. Purpose: To evaluate the effect of etched dentin treatment with proanthocyanidin (PA) in short periods of time on the inhibition of dentin metalloproteinases (MMP) activity in situ. Material and method: Forty dentin specimens (1x1x6mm) were obtained from sound third molars and divided into 4 groups (n=10). The specimens were etched with 37% phosphoric acid for 15s and rinsed in deionized water. Then they were treated with the following solutions: water, 5% PA for 5s, 15s or 30s. The total MMP activity was analyzed by a colorimetric test (SensoLyte®). Absorbance data (412nm) were submitted to ANOVA and Tukey tests (alfa =0.05). Result: All treatment periods were able to reduce the total activity of MMPs. The best results were observed for dentine treated with PA for 15s (63.1% reduction) and 30s (70.2%). Treatment for 5s was capable of inhibiting only 39.9% of the total MMP activity. Conclusion: Application of PA on the etched dentin in extremely short periods of time reduced the MMPs activity of dentin, even after 5s. However, the best results were obtained for the longer periods.

Transdentinal cytotoxicity of glutaraldehyde on odontoblast-like cells.

OBJECTIVES: This study investigated the transdentinal cytotoxicity of glutahaldehyde-containing solutions/materials on odontoblast-like cells. METHODS: Dentin discs were adapted to artificial pulp chambers. MDPC-23 cells were seeded on the pulpal side of the discs and the occlusal surface was treated with the following solutions: water, 2% glutaraldehyde (GA), 5% GA, 10% GA, Gluma Comfort Bond+Desensitizer (GCB+De) or Gluma Desensitizer (GDe). Cell viability and morphology were assessed by the Alamar Blue assay and SEM. The eluates were collected and applied on cells seeded in 24-well plates. After 7 or 14 days the total protein (TP) production, alkaline phosphatase activity (ALP) and deposition of mineralized nodules (MN) were evaluated. RESULTS: Data were analyzed by Kruskal-Wallis and Mann-Whitney tests (p<0.05). GA solutions were not cytotoxic against MDPC-23. GCB+De (85.1%) and GDe (77.2%) reduced cell viability as well as TP production and ALP activity at both periods. After 14 days, GCB+De and GDe groups produced less MN. Affected MDPC-23 presented deformation of the cytoskeleton and reduction of cellular projections. CONCLUSIONS: The treatment with 2.5%, 5% and 10% GA was not harmful to odontoblast-like cells. Conversely, when GA was combined with other components like HEMA, the final material became cytotoxic. CLINICAL SIGNIFICANCE: Glutaraldehyde has been used to decrease dentin hypersensitivity. This substance is also capable of preventing resin-dentin bond degradation by cross-linking collagen and MMPs. This study showed that GA might be safe when applied on acid etched dentin. However, when combined with HEMA the product becomes cytotoxic.

Immediate human pulp response to ethanol-wet bonding technique.

OBJECTIVES: To evaluate the short-term response of human pulps to ethanol-wet bonding technique. METHODS: Deep class V cavities were prepared on 17 sound premolars and divided into three groups. After acid-etching, the cavities from groups 1 (G1) and 2 (G2) were filled with 100% ethanol or distilled water, respectively, for 60 s before the application of Single Bond 2. In group 3 (G3, control), the cavity floor was lined with calcium hydroxide before etching and bonding. All cavities were restored with resin composite. Two teeth were used as intact control. The teeth were extracted 48h after the clinical procedures. From each tooth serial sections were obtained and stained with haematoxylin and eosin (H/E) and Masson's trichrome. Bacteria microleakage was assessed using Brown & Brenn. All sections were blindly evaluated for five histological features. RESULTS: Mean remaining dentine thickness was 463±65µm (G1); 425±184µm (G2); and 348±194µm (G3). Similar pulp reactions followed ethanol- or water-wet bonding techniques. Slight inflammatory responses and disruption of the odontoblast layer related to the cavity floor were seen in all groups. Stained bacteria were not detected in any cavities. Normal pulp tissue was observed in G3 except for one case. CONCLUSIONS: After 48h, ethanol-wet bonding does not increase pulpal damage compared to water-wet bonding technique. CLINICAL SIGNIFICANCE: Ethanol-wet bonding may increase resin-dentine bond durability. This study reported the in vivo response of human pulp tissue when 100% ethanol was applied previously to an etch-and-rinse simplified adhesive system.

Esthetic dental anomalies as motive for bullying in schoolchildren.

Facial esthetics, including oral esthetics, can severely affect children's quality-of-life, causing physical, social and psychological impairment. Children and adolescents with esthetic-related dental malformations are potential targets for bullies. This study was aimed to present and discuss patients who suffered from bullying at school and family environment due to esthetic-related teeth anomalies. Providing an adequate esthetic dental treatment is an important step in their rehabilitation when the lack of esthetic is the main source of bullying. After dental treatment, we noted significant improvement in self-esteem, self-confidence, socialization and academic performance of all patients and improvement in parental satisfaction regarding the appearance of their children. It is imperative that both family and school care providers be constantly alert about bullying in order to prevent or interrupt aggressive and discriminatory practices against children and adolescents. Clearly, dental anomalies may be a motive for bullying.

Effect of time between adhesive application and photoactivation on adhesion and collagen exposure.

PURPOSE: To evaluate the influence of the time elapsed between the application and photoactivation of Single Bond 2 (SB) on microtensile bond strength (µTBS) and collagen exposure at the adhesive interface produced in the presence of intrapulp pressure. METHODS: Dentin occlusal surfaces were prepared in 72 molars, and divided into eight groups (n = 9). After acid etching, SB was applied, with or without simulated intrapulp pressure, and remained undisturbed for 0, 20, 40 or 60 seconds, before photoactivation. Three teeth/group were processed for staining with Goldner trichrome and evaluation of the thickness of exposed collagen zone (CZ) at the base of the hybrid layer. Composite resin build-ups were placed on the remaining six prepared teeth prior to sectioning for microtensile testing. Data were submitted to ANOVA and Tukey tests (α = 0.05). RESULTS: In the absence of pressure, immediate photoactivation resulted in the lowest µTBS, while the other groups did not differ among them. Under intrapulp pressure, the lowest values were observed after 60 seconds. There was no difference in the thickness of the exposed collagen zone among the groups without pressure. However, thicker layers were recorded in the presence of pressure after 40 and 60 seconds. Waiting 60 seconds between application and photoactivation of SB significantly reduced resin-dentin bond strength when pulpal pressure was simulated.

Effect of reducing acid etching time on bond strength to noncarious and caries-affected primary and permanent dentin.

PURPOSE: The purpose was to evaluate the effect of acid etching time on the bond strength of a simplified etch-and-rinse adhesive system to noncarious and caries-affected dentin of primary and permanent teeth. METHODS: Twenty-four extracted primary and permanent teeth were divided into three groups, according to the acid etching time. Four teeth from each group were exposed to a microbiological caries-inducing protocol. After caries removal, noncarious and caries-affected dentin surfaces were etched with 37 percent phosphoric acid for five, 10, or 15 seconds prior to the application of Prime & Bond NT adhesive. Crowns were restored with resin composite and prepared for microtensile testing. Data were submitted to Kruskal-Wallis and Mann-Whitney tests (α=0.05). RESULTS: Higher bond strengths were obtained for noncarious dentin vs. caries-affected dentin for both primary and permanent teeth. Reducing the acid etching time from 15 to five seconds did not affect the bond strength to caries-affected or noncarious dentin in primary teeth. For permanent teeth, lower bond strength values were observed when the noncarious dentin was etched for five seconds, while no difference was seen between 10 and 15 seconds. CONCLUSIONS: For Prime & Bond NT, the etching of dentin for five seconds could be recommended for primary teeth, while 10 seconds would be the minimum time for permanent teeth.

Citotoxicidade transdentinária e efeito da carbodiimida (EDC) na biomodificação do colágeno dentinário e na degradação da interface adesiva / Trandentinal cytotoxicity and effect of carbodiimide (EDC) on dentin collagen biomodification and adhesive interface degradation

O objetivo geral deste trabalho foi avaliar a citotoxicidade transdentinária da carbodiimida (EDC), bem como sua influência na degradação do colágeno dentinário e na estabilidade da união resina-dentina. No estudo 1, células MDPC23 foram plantadas na superfície pulpar de discos de dentina e a superfície oclusal foi tratada por 60s com uma das seguintes soluções: sem tratamento; EDC 0,1M; 0,3M ou 0,5M; glutaraldeído 5% (GA); tampão Sorensen ou H2O2 29%. A viabilidade e a morfologia celular foram analisadas pelos testes de MTT, Live/dead, produção de proteína total (PT), de colágeno e MEV. Os dados foram analisados pelos testes de Kruskal-Wallis e Mann-Whitney (p<0,05). O GA promoveu aumento do metabolismo celular. A morte por necrose e a morfologia celular não foram influenciadas pelos agentes cross-linkers. Não houve redução na produção de PT e colágeno após 7 dias. Para o estudo 2, espécimes de dentina foram completamente desmineralizados e a variação do módulo de elasticidade (E), inibição de MMP, perda de massa, liberação de hidroxiprolina (HYP) e degradação térmica do colágeno (DTC) foram analisados após tratamento com uma das seguintes soluções por 30s ou 60s: água deionizada (controle); EDC 0,5M; EDC 1M; EDC 2M e GA 10%. Os dados referentes ao E, atividade de MMP e liberação de HYP foram submetidos aos testes de Wilcoxon e KruskalWallis ou Mann-Whitney. Os valores de perda de massa e DT foram analisados pelos testes de ANOVA e Tukey (p<0,05). Os melhores resultados quanto ao E foram observados para o GA. Todos os cross-linkers reduziram a atividade de MMP e a liberação de HYP e aumentaram a temperatura de DT do colágeno. No estudo 3, sessenta palitos de dentina foram divididos em 6 grupos de acordo com a solução de tratamento: água deionizada (controle); EDC 0,1M; EDC 0,5M; EDC 0,5M + HEMA 35%; proantocianidina 5% (PA) ou clorexidina (CHX) 2%. Após condicionamento ácido os palitos foram tratados por 60s. A atividade de MMP foi analisada por ensaio colorimétrico. Os dados expressos em valores de absorbância e em equivalentes a atividade de MMP-9 foram submetidos aos testes de Kruskall-Wallis e Mann-Whitney (p<0.05). Todas as soluções inibiram a atividade de MMP. O EDC 0,5M e sua mistura com HEMA obtiveram os melhores resultados. Finalmente, no estudo 4, superfícies planas de dentina foram condicionadas e tratadas por 30 ou 60s com solução de EDC 0,5M ou água deionizada. Após o tratamento, o adesivo Single Bond 2 (SB2) foi aplicado e as coroas reconstruídas em resina composta. Espécimes foram produzidos e submetidos aos testes de microtração e nanoinfiltração após 24h, 6 ou 12 meses em saliva artificial. Os testes de ANOVA e Tukey (p<0.05) foram aplicados. O tratamento da dentina com EDC preveniu a redução da RU e o aumento da nanoinfiltração após 12 meses de armazenamento. Desta forma, pôde ser concluído que o pré-tratamento da dentina com agentes cross-linkers não exerceu efeito citotóxico trandentinário sobre células odontoblastóides, favoreceu as propriedades mecânicas do colágeno, foi capaz de inibir MMPs e prevenir a degradação da interface adesiva

The purpose of this study was to evaluate the trandentinal cytotoxicity of carbodiimide (EDC), as well as its influence on dentinal collagen degradation and stability of resin-dentin bonds. In the first experiment, MDPC-23 cells were seeded on the pulp surface of the disks and one of the following solutions was applied on the occlusal surface for 60s: no treatment (negative control), 0.1M, 0.3M or 0.5M EDC; 5% glutaraldehyde (GA); Sorensen buffer or 29% hydrogen peroxide (positive control). Cell viability and morphology were analyzed by MTT, Live/Dead assays, total protein (TP) and collagen production and SEM. Data were analyzed by Kruskal-Wallis and Mann-Whitney tests (p<0.05). Only GA increased cellular metabolism. Cell death by necrosis and cell morphology were not affected by the cross-linker agents. There was no reduction in TP and collagen production after 7 days. For the second experiment, dentin beams were completely demineralized and the variation in elastic modulus (E), MMP activity, dry mass loss, hydroxyproline release (HYP) and collagen thermal degradation (CTD) were analyzed after the dentin treatment for 30s or 60s with the following solutions: water; 0.5M; 1M or 2M EDC and 10% GA. Data from E and MMP activity and HYP release were submitted to Wilcoxon and Kruskal-Wallis or Mann-Whitney tests. Dry mass loss and CTD data were analyzed by ANOVA and Tukey's tests (p>0.05). GA group obtained the highest E values. All cross-linking agents decreased MMP activity and HYP release and increased CTD. In the third experiment, sixty dentin beams were randomly divided into 6 groups according to the treatment solution: deionized water (control), 0.1M EDC, 0.5M EDC, 0.5M EDC+35% HEMA, 5% proanthocyanidin (PA) or 2% chlorhexidine (CHX). The beams were acid etched and treated for 60s. The total MMP activity was analyzed by a colorimetric assay (Sensolyte®). Data were expressed as absorbance values at 412nm and MMP-9 equivalents and subjected to Kruskal-Wallis and MannWhitney tests (p<0.05). All cross-liking solutions inhibited MMPs. The 0.5 M EDC solution and its mixture with HEMA had the highest inhibition values. Finally, in experiment 4, flat dentin surfaces were etched and treated for 30 or 60s with 0.5M EDC or deionized water. After treatment, Single Bond 2 (SB2) was applied and the crowns were reconstructed with composite resin. Dentin specimens were produced and submitted to microtensile and nanoleakage tests after 24h, 6 or 12 months in artificial saliva. Bond strength (BS) data (MPa) were analyzed by ANOVA and Tukey tests (p<0.05). The dentin treatment with EDC did not affect SB2 immediate BS and prevented the degradation of the adhesive interface, even after 12 months of saliva storage. Thus, the dentin treatment with cross-linking agents did not exert transdentinal cytotoxic effects on odontoblastlike cells, increased collagen mechanical properties, was able to inhibit MMPs and prevent the degradation of the adhesive interface

Effect of acid etching time on demineralization of primary and permanent coronal dentin.

PURPOSE: To determine the effect of acid etching time on dentin calcium solubilization and to compare the solubilization rate of primary and permanent dentin when etched for the same period of time. METHODS: Flat coronal dentin surfaces were produced on primary (n=8) and permanent (n=8) sound teeth. Three 2 mm-diameter areas were delimited on each surface and assigned to 5, 10 or 15 seconds of acid etching. Five microliters of 35% H3PO4 were applied on each area during the preset times, and 4 microL were retrieved for analysis. The amount of calcium was determined colorimetrically using Arsenazo III reagent and expressed as microg Ca/mm2 of dentin. Data were subjected to two-way ANOVA, Tukey's test and linear regression (alpha=5%). RESULTS: For both primary and permanent dentin, a statistically significant correlation was observed between acid etching time and calcium concentration (primary dentin R2 = 0.79; permanent dentin R2 = 0.73). Mean (sd) values of dissolved primary dentin after 5, 10 and 15 seconds were 1.00 (0.25), 1.74 (0.11) and 2.30 (0.42), whereas for permanent dentin the values were 0.47 (0.13), 0.89 (0.36) and 1.38 (0.20) microg Ca/mm2, respectively. Greater calcium solubilization (P<0.05) was detected for primary dentin than for permanent dentin after all acid etching times.

Tempo de condicionamento da dentina hígida e afetada por cárie de dentes decíduos e permanentes: efeito na desmineralização do substrato, na produção e na resistência da união resina-dentina / Acid etching time of sound and caries-affected dentin of deciduos and permanent teeth: effect on demineralization of the substrate, production and adhesive performance of resin-dentin bonds

O objetivo geral desse trabalho, dividido em três estudos, foi avaliar a influência do tempo de condicionamento ácido na desmineralização da dentina, na qualidade e no desempenho adesivo imediato da união resina-dentina de dentes decíduos e permanentes. No estudo 1, superfícies planas de dentina foram produzidas em molares decíduos e pré-molares (n=8). Sobre cada superfície, três áreas foram delimitadas e condicionadas com ácido fosfórico por 5, 10 ou 15 s. Decorrido o período pré-estabelecido de condicionamento, o ácido foi coletado e a concentração de cálcio dissolvido da dentina (ug Ca/mm2) foi determinada por microcolorimetria. Os dados foram analisados pela aplicação dos testes de ANOVA, Tukey e análise de regressão linear (α=0,05). Correlação positiva significante foi observada entre a concentração de cálcio e o tempo de condicionamento ácido para ambos, dentes decíduos e permanentes. Para todos os tempos de condicionamento, quantidades estatisticamente superiores de íons cálcio foram detectadas para a dentina de dentes decíduos em comparação a dentina de dentes permanentes. Foi concluído que a dentina de dentes decíduos é mais facilmente desmineralizada do que a dentina de dentes permanentes, e que essa desmineralização foi maior em função do aumento do tempo de aplicação do ácido. No estudo 2, superfícies planas de dentina também foram produzidas em outros 8 molares decíduos e 8 pré-molares hígidos. As superfícies foram divididas ao meio no sentido vestíbulo-lingual por meio de uma canaleta produzida com um disco diamantado. Aleatoriamente, cada metade foi condicionada com ácido fosfórico por 15 ou 5 s e os sistemas adesivos Prime&Bond NT ou Prime&Bond 2.1 foram aplicados. Espécimes envolvendo a interface de união foram produzidos e preparados em laboratório para coloração com tricrômico de Goldner. A espessura da zona de colágeno exposta na base da camada híbrida foi mensurada em microscopia óptica e os dados submetidos a análise de variância a dois critérios fixos e testes complementares de Tukey (α=0,05). Em geral, maior exposição de fibrilas de colágeno foi observada quando o mesmo substrato foi condicionado por 15 s em comparação ao condicionamento por 5 s. Maior exposição de colágeno também foi observada em dentes decíduos em comparação a dentes permanentes, entretanto apenas quando o sistema Prime&Bond 2.1 foi utilizado. Pôde ser concluído que o aumento do tempo de condicionamento ácido dificulta a infiltração completa da zona de dentina desmineralizada favorecendo a manutenção de fibrilas de colágeno expostas na base da camada híbrida. Por fim, no estudo 3, das 24 superfícies planas de dentina produzidas em molares decíduos e 24 em pré-molares, metade foi submetida a um protocolo artificial de cárie. Nos dentes cariados, a dentina infectada foi completamente removida deixando como substrato para adesão a dentina afetada por cárie. Sobre esse substrato e também sobre as superfícies mantidas hígidas, foi realizado o condicionamento com ácido fosfórico por 5, 10 ou 15 s, seguido da aplicação do sistema adesivo Prime&Bond NT. Espécimes (0,81 mm2 ) foram produzidos para o ensaio mecânico de microtração e os dados de resistência de união (RU) foram analisados pela aplicação dos testes de Kruskal-Wallis e Mann-Whitney (α=0,05). A redução do tempo de condicionamento afetou negativamente os valores de resistência de união apenas para a dentina hígida de dentes permanentes, enquanto nenhum efeito negativo foi observado para a dentina de dentes decíduos, hígida ou afetada por cárie, e para a dentina afetada por cárie de dentes permanentes. Em conclusão, uma vez que dentina hígida e afetada por cárie coexistem na maioria dos preparos cavitários, o condicionamento ácido por no mínimo 10 s deve ser recomendado para dentes permanentes, enquanto períodos inferiores podem ser indicados para dentes decíduos

The aim of this work, divided into three studies, was to evaluate the influence of acid etching time on dentin demineralization, quality and immediate adhesive performance of resin-dentin bonds produced in deciduous and permanent teeth. In the first study, flat dentin surfaces were produced in sound primary molars (n=8) and premolars (n=8). On each surface, three circular areas were defined and etched with phosphoric acid for 5, 10 or 15 s. After the predetermined period of etching, the acid was collected and the calcium concentration (ug Ca/mm2) was determined by microcolorimetry. The data were analyzed by ANOVA, Tukey and linear regression tests (α=0.05). Significant positive correlation was observed between calcium concentration and etching time for both deciduous and permanent teeth. For all times of conditioning, statistically higher amounts of calcium ions were removed from dentin of primary teeth compared to dentin of permanent teeth. It was concluded that the primary dentin was more prone to demineralization by phosphoric acid than permanent dentin, and that the extent of demineralization increased as a function of acid etching time. In the second study, flat dentin surfaces were also produced in additional 8 primary molars and 8 premolars. The surfaces were divided into mesial and distal halves through a shallow notch produced with a diamond disc. Randomly, each half was conditioned with 35% phosphoric acid for 15 or 5 s and the adhesive systems Prime & Bond NT or Prime & Bond 2.1 were applied. Specimens involving the bonded interface were produced and processed for staining with Goldner's trichrome. The thickness of the collagen zone exposed at the base of the hybrid layer (ZC) was measured using optical microscopy. Data were submitted to threeway analysis of variance and Tukey's test (α=0.05). Overall, thicker ZC were observed when the same substrate was etched for 15 s compared to 5 s. Also, thicker ZC were seen for primary dentin when etched for the same length of time than the permanent dentin, reaching statistical significance for Prime&Bond 2.1. It was concluded that extending the etching time from 5 s to 15 s was detrimental to the monomeric infiltration of the demineralized dentin, favoring the maintenance of exposed collagen fibrils at the base of the hybrid layer. In the last study, flat dentin surfaces were produced in 24 primary molars and 24 premolars. Half of them were subjected to an artificial caries protocol using S. Mutans. In carious teeth, the infected dentin was completely removed leaving as a substrate for adhesion the caries-affected dentin. On this substrate and also on the noncarious dentin, 35% phosphoric acid was applied for 5, 10 or 15 s, followed by application of Prime & Bond NT. Beam shaped specimens (0.81 mm2) were produced for microtensile test and bond strength data were analyzed by KruskalWallis and Mann-Whitney tests (α=0.05). The reduction in acid etching time negatively affected bond strength only for sound dentin of permanent teeth. No negative effect was observed for primary dentin, sound or caries-affected, and caries-affected permanent dentin. In conclusion, since sound and caries-affected dentin coexist in the majority of the cavity preparations, at least 10 s of acid etching should be recommended for permanent teeth, while shorter periods could be indicated for primary teeth