Effect of crosslinkers on bond strength stability of fiber posts to root canal dentin and in situ proteolytic activity.

STATEMENT OF PROBLEM: Improved stability of the adhesive interface can be obtained using crosslinkers. However, research on the use of crosslinkers in root dentin is lacking. PURPOSE: The purpose of this in vitro study was to evaluate the effect of crosslinkers on the proteolytic activity of root dentin and on the bond strength of resin-cemented fiber posts. MATERIAL AND METHODS: Single root canals were obtained from premolars (n=48) and endodontically treated before being divided into 4 groups: deionized water (control), 0.5 mol/L carbodiimide, 5% proanthocyanidin, or 5% glutaraldehyde. After removing the canal sealer, the dentin was etched with phosphoric acid, followed by water rinsing and the application of the crosslinkers for 60 seconds. Fiber posts were cemented using an adhesive (Single Bond 2) and resin cement (RelyX ARC). The roots were then transversally sectioned to obtain 1 mm thick specimens from the cervical, middle, and apical thirds and then aged for 24 hours or 9 months. Nine roots per group were used for the push-out test and 3 for determining the proteolytic activity of the root dentin by in situ zymography. Bond strength data were submitted to a mixed-model ANOVA and Bonferroni tests (α=.05). RESULTS: Only proanthocyanidin negatively affected the 24-hour bond strength. After 9 months, a significant decrease in bond strength was seen for all groups, except for the crosslinked treated specimens from the cervical third of the root canal. Intense gelatinolytic activity was detected in the control group after 24 hours but was inhibited in the crosslinker-treated groups. Proteolytic activity was also not detected after 9 months for the groups treated with the crosslinkers, irrespective of the root canal third. Conversely, proteolytic activity increased for the specimens from the control group. CONCLUSIONS: Although no proteolytic activity was detected in the hybrid layers along the entire root canal, dentin biomodification with crosslinkers was effective in preventing bond strength loss only in the cervical third.

Cross-linked dry bonding: A new etch-and-rinse technique.

OBJECTIVE: To determine if acid-etched, cross-linked dentin can be dehydrated without lowering bond strength below that of cross-linked wet-bonded dentin in vitro. METHODS: Using extracted human third molars, control acid-etched dentin was bonded with Single Bond Plus, using either the wet- or dry-bonding technique. Experimental acid-etched dentin was treated with 5mass% grape seed extract (GSE) in different solvents for 1min before undergoing wet vs dry resin-dentin bonding with Single Bond Plus. Completely demineralized dentin beams were treated with 5% GSE for 0, 1 or 10min, before measuring stiffness by 3-point flexure. Other completely demineralized beams were treated similarly and then incubated in buffer for 1 week to measure the collagen solubilization by endogenous dentin proteases. RESULTS: 24h microtensile bond strengths (µTBS) in wet and dry controls were 53.5±3.6 and 9.4±1.8MPa, respectively (p<0.05). 5% GSE in water gave µTBS of 53.7±3.4 and 39.1±9.7MPa (p<0.05), respectively, while 5% GSE in ethanol gave µTBS of 51.2±2.3 and 35.3±2.0MPa (p<0.05). 5% GSE in 5% EtOH/95% water gave wet and dry µTBS of 53.0±2.3 and 55.7±5.1MPa (p>0.05). Cross-linking demineralized dentin with 5% GSE increased stiffness of dentin and decreased collagen degradation (p<0.05). SIGNIFICANCE: 5% GSE pretreatment of acid-etched dentin for 1min permits the dentin to be completely air-dried without lowering bond strength.

Effects of a Dicalcium and Tetracalcium Phosphate-Based Desensitizer on In Vitro Dentin Permeability.

The present study evaluated the effectiveness of a dicalcium and tetracalcium phosphate-based desensitizer in reducing dentin permeability in vitro. Dentin fluid flow was measured before and after treatment of dentin with patent dentinal tubules using 1 or 3 applications of the dicalcium and tetracalcium phosphate containing agent TeethmateTM (TM) and comparing the results with two sodium fluoride varnishes VellaTM (VLA) and VanishTM (VAN), after storage in artificial saliva for 24 h, 48 h and 7 days. Significant differences were observed among the 4 methods employed for reducing dentin permeability (p < 0.001) and the 3 post-treatment times (p < 0.001). VLA and VAN never achieved 50% permeability reductions consistently in any of the 3 time periods. Only the calcium phosphate-based desensitizer applied for 3 times consistently reduced dentin permeability by 50% after 24 h. When applied once, the permeability reduction of TM increased progressively over the 3 time periods. After 7 days, only one and three applications of the calcium phosphate-based desensitizer consistently reduced dentin permeability by more than 50%. Permeability reductions corresponded well with scanning electron microscopy examination of dentinal tubule orifice occlusion in dentin specimens treated with the agents. Overall, the dicalcium and tetracalcium phosphate-based desensitizer is effective in reducing dentin permeability via a tubule occlusion mechanism. The ability of the agent to reduce dentin permeability renders it to be potentially useful as a clinical dentin desensitizing agent, which has to be confirmed in future clinical studies. By contrast, the two sodium fluoride varnishes are not effective in dentin permeability reduction and should be considered as topical fluoride delivering agents rather than tubular orifice-blocking agents.

Efeito citotóxico de géis clareadores com diferentes concentrações de peróxido de hidrogênio aplicados diretamente sobre a dentina / Cytotoxic effects of a bleaching gel with different concentrations of hydrogen peroxide applied directly on dentin

Objetivo: Avaliar os efeitos citotóxicos de agentes clareadores com diferentes concentrações de PH sobre células odontoblastóides, quando aplicados diretamente sobre a superfície de dentina humana. Material e método: Cinquenta discos de dentina (0,5 mm de espessura) foram adaptados em câmaras pulpares artificiais (CPAs) e células MDPC-23 foram semeadas na superfície pulpar dos discos. Cinco grupos (n=10) foram estabelecidos: G1: 7,5% PH; G2: 20% PH; G3: 35% PH; G4: gel sem PH; G5: DMEN (controle). Os produtos foram aplicados na superfície oclusal dos discos por 2x de 15 minutos. A viabilidade (ensaio de MTT) e a morfologia celular (MEV) foram avaliadas imediatamente após o clareamento. Os dados de viabilidade celular foram submetidos ao teste de Kruskal-Wallis e Mann-Whitney (α=0,05). Resultados: Redução significante na viabilidade celular em relação ao controle (G5) foi observada para todas as concentrações de PH (p<0,05), associada a intensas alterações na morfologia celular. Entretanto, nenhuma diferença significante foi observada entre as três concentrações de PH. Também, não houve diferença estatística entre o grupo controle e o grupo gel sem PH (G5 e G4). Conclusão: Todas as concentrações de PH causaram efeitos citotóxicos de severos sobre as células MDPC-23, quando aplicados diretamente sobre a dentina. Entretanto, a intensidade do efeito tóxico não foi influenciada pela concentração de PH no agente clareador. Relevância clínica: Apesar das limitações deste estudo in vitro, os resultados indicam que o clareamento dental não deve ser realizado diretamente em áreas com exposição da dentina.

Objective: To evaluate the cytotoxic effects of bleaching gels with different concentrations of hydrogen peroxide (HP) on odontoblast-like cells, when applied directly on dentin. Material and method: Fifty dentin discs (0.5 mm thick) were adapted in artificial pulp chambers (APC) and MDPC-23 cells were seed on the pulpal side. The discs were divided into 5 groups (n=10): G1: HP 7.5%; G2: HP 20%; G3: HP 35%; G4: gel with no HP; and G5: no treatment (control). The gels were applied on the occlusal side of the discs for 2x of 15 min. Cellular viability (MTT assay) and morphology (SEM) were analyzed immediately after the bleaching procedure. Data of cellular viability were submitted to Kruskal-Wallis and Mann-Whitney tests (α=0.05). Results: Significant reduction in cellular viability was seen for all HP concentrations in comparison to the control (G5). However, no statistical significant difference was seen among the concentrations of HP. Likewise, there was no statistical difference between the control group (G5) and the group where the gel with no HP was applied (G4). Conclusion: All HP concentrations caused severe cytotoxic effects on the odontoblast-like cells when applied directly on dentin. However, the intensity of the cytotoxic effect was not influenced by the concentration of the HP included in the bleaching gel. Clinical significance: Within the limitations of this in vitro study, the results strongly indicate that dental bleaching procedures should not be performed directly on areas of dentin exposure.

Water distribution in dentin matrices: bound vs. unbound water.

OBJECTIVE: This work measured the amount of bound versus unbound water in completely-demineralized dentin. METHODS: Dentin beams prepared from extracted human teeth were completely demineralized, rinsed and dried to constant mass. They were rehydrated in 41% relative humidity (RH), while gravimetrically measuring their mass increase until the first plateau was reached at 0.064 (vacuum) or 0.116 gH2O/g dry mass (Drierite). The specimens were then exposed to 60% RH until attaining the second plateau at 0.220 (vacuum) or 0.191 gH2O/g dry mass (Drierite), and subsequently exposed to 99% RH until attaining the third plateau at 0.493 (vacuum) or 0.401 gH2O/g dry mass (Drierite). RESULTS: Exposure of the first layer of bound water to 0% RH for 5 min produced a -0.3% loss of bound water; in the second layer of bound water it caused a -3.3% loss of bound water; in the third layer it caused a -6% loss of bound water. Immersion in 100% ethanol or acetone for 5 min produced a 2.8 and 1.9% loss of bound water from the first layer, respectively; it caused a -4 and -7% loss of bound water in the second layer, respectively; and a -17 and -23% loss of bound water in the third layer. Bound water represented 21-25% of total dentin water. Chemical dehydration of water-saturated dentin with ethanol/acetone for 1 min only removed between 25 and 35% of unbound water, respectively. SIGNIFICANCE: Attempts to remove bound water by evaporation were not very successful. Chemical dehydration with 100% acetone was more successful than 100% ethanol especially the third layer of bound water. Since unbound water represents between 75 and 79% of total matrix water, the more such water can be removed, the more resin can be infiltrated.

Methods to evaluate and strategies to improve the biocompatibility of dental materials and operative techniques.

OBJECTIVE: The general aim of this article is to describe the state-of-the-art of biocompatibility testing for dental materials, and present new strategies for improving operative dentistry techniques and the biocompatibility of dental materials as they relate to their interaction with the dentin-pulp complex. METHODS: The literature was reviewed focusing on articles related to biocompatibilty testing, the dentin-pulp complex and new strategies and materials for operative dentistry. For this purpose, the PubMed database as well as 118 articles published in English from 1939 to 2014 were searched. Data concerning types of biological tests and standardization of in vitro and in vivo protocols employed to evaluate the cytotoxicity and biocompatibility of dental materials were also searched from the US Food and Drug Administration (FDA), International Standards Organization (ISO) and American National Standards Institute (ANSI). RESULTS: While there is an ongoing search for feasible strategies in the molecular approach to direct the repair or regeneration of structures that form the oral tissues, it is necessary for professionals to master the clinical therapies available at present. In turn, these techniques must be applied based on knowledge of the morphological and physiological characteristics of the tissues involved, as well as the physical, mechanical and biologic properties of the biomaterials recommended for each specific situation. Thus, particularly within modern esthetic restorative dentistry, the use of minimally invasive operative techniques associated with the use of dental materials with excellent properties and scientifically proved by means of clinical and laboratory studies must be a routine for dentists. This professional and responsible attitude will certainly result in greater possibility of achieving clinical success, benefiting patients and dentists themselves. SIGNIFICANCE: This article provides a general and critical view of the relations that permeate the interaction between dental materials and the dentin-pulp complex, and establish real possibilities and strategies that favor biocompatibility of the present and new products used in Dentistry, which will certainly benefit clinicians and their patients.

Inhibition of endogenous human dentin MMPs by Gluma.

OBJECTIVE: The objective of this study was to determine if Gluma dentin desensitizer (5.0% glutaraldehyde and 35% HEMA in water) can inhibit the endogenous MMPs of dentin matrices in 60 s and to evaluate its effect on dentin matrix stiffness and dry mass weight. METHODS: Dentin beams of 2 mm×1 mm×6 mm were obtained from extracted human third molars coronal dentin. To measure the influence of Gluma treatment time on total MMP activity of dentin, beams were dipped in 37% phosphoric acid (PA) for 15 s and rinsed in water. The acid-etched beams were then dipped in Gluma for 5, 15, 30 or 60 s, rinsed in water and incubated into SensoLyte generic MMP substrate (AnaSpec, Inc.) for 60 min. Controls were dipped in water for 60 s. Additional beams of 1 mm×1 mm×6 mm were completely demineralized in 37% PA for 18 h, rinsed and used to evaluate changes on the dry weight and modulus of elasticity (E) after 60 s of Gluma treatment followed by incubation in simulated body fluid buffer for 0, 1 or 4 weeks. E was measured by 3-pt flexure. RESULTS: Gluma treatment inhibited total MMP activity of acid-etched dentin by 44, 50, 84, 86% after 5, 15, 30 or 60 s of exposure, respectively. All completely demineralized dentin beams lost stiffness after 1 and 4 weeks, with no significant differences between the control and Gluma-treated dentin. Gluma treatment for 60 s yielded significantly less dry mass loss than the control after 4 weeks. SIGNIFICANCE: The use of Gluma may contribute to the preservation of adhesive interfaces by its cross-linking and inhibitory properties of endogenous dentin MMPs.

Stabilization of dentin matrix after cross-linking treatments, in vitro.

OBJECTIVES: To evaluate the effect of EDC on elastic modulus (E), MMPs activity, hydroxyproline (HYP) release and thermal denaturation temperature of demineralized dentin collagen. METHODS: Dentin beams were obtained from human molars and completely demineralized in 10 wt% H3PO4 for 18 h. The initial E and MMP activity were determined with three-point bending and microcolorimetric assay, respectively. Extra demineralized beams were dehydrated and the initial dry mass (DM) was determined. All the beams were distributed into groups (n=10) and treated for 30 s or 60 s with: water, 0.5 M, 1 M or 2 M EDC or 10% glutaraldehyde (GA). After treatment, the new E and MMP activity were redetermined. The beams submitted to DM measurements were storage for 1 week in artificial saliva, after that the mass loss and HYP release were evaluated. The collagen thermal denaturation temperature (TDT) was determined by DSC analysis. Data for E, MMP activity and HYP release were submitted to Wilcoxon and Kruskal-Wallis or Mann-Whitney tests. Mass loss and TDT data were submitted to ANOVA and Tukey tests at the 5% of significance. RESULTS: EDC was able to significantly increase collagen stiffness in 60s. 10% GA groups obtained the highest E values after both 30 and 60s. All cross-linking agents decreased MMP activity and HYP release and increased TDT temperature. Significant differences were identified among EDC groups after 30 or 60 s of cross-linking, 1M or 2M EDC showed the lowest MMP activity. SIGNIFICANCE: Cross-linking agents are capable of preventing dentin collagen degradation. EDC treatment may be clinically useful to increase resin-dentin stability.

Influência da clorexidina na capacidade de umectablidade da dentina hígida e afetada por cárie por um sistema adesivo / Influence of chlorhexidine on the wettability of sound and caries-affected dentin by an adhesive system

Objetivo: Avaliar o efeito da clorexidina (CLX) na umectabilidade da dentina hígida e afetada por cárie por um sistema adesivo convencional simplificado. Material e Método: Foram preparadas superfícies planas de dentina em 60 molares hígidos, dos quais 30 foram artificialmente cariados. Para cada condição de substrato, hígido e afetado por cárie, as superfícies foram divididas em 3 grupos (n=10): com smear layer (SL), sem SL impregnada com água e sem SL impregnada com CLX. A remoção da SL foi realizada pela aplicação de ácido fosfórico por 15 s. Sobre a dentina desmineralizada foram aplicados 20 uL de água destilada ou digluconato de CLX a 2% por 60 s. Em seguida, uma gota do sistema Single Bond 2 foi depositada sobre cada superfície. Ângulos de contato entre a superfície da dentina e o adesivo foram mensurados por meio de um goniômetro e os dados submetidos aos testes de ANOVA e Tukey (?=0,05). Resultados: Maiores ângulos de contato foram obtidos sobre a dentina hígida em comparação a afetada por cárie (p<0,05), independente do tratamento da superfície. Para ambos os substratos, ângulos de contato estatisticamente superiores foram obtidos para a dentina coberta com SL (p<0,05). A remoção da SL resultou em redução significante dos ângulos (p<0,05) e nenhuma diferença foi encontrada entre os ângulos produzidos sobre a dentina desmineralizada impregnada por água ou por CLX (p>0,05). Conclusão: A umectabilidade da dentina afetada por cárie foi maior do que a da dentina hígida, sendo que a mesma não foi influenciada pela aplicação de clorexidina.

Aim: To evaluate the effect of chlorhexidine (CHX) on the wettability of sound and caries affected dentin by a simplified adhesive system. Material and Methods: Flat coronal dentin surfaces were produced on 60 sound molars, 30 of which were artificially decayed. The teeth were divided randomly into 3 groups (n = 10) with smear layer (SL), without SL impregnated with water and without SL impregnated with chlorhexidine. The SL removal was performed by phosphoric acid etching for 15 s. 20 uL of distilled water or 2% chlorhexidine digluconate were applied on the demineralized dentin for 60 s. Then, a drop of Single Bond 2 was deposited on each surface. Contact angles between dentin surface and adhesive was measured by means of a goniometer and data were submitted to ANOVA and Tukey tests (? = 0.05). Results: Higher contact angles were obtained on sound versus caries affected dentin (p <0.05), regardeless of the surface treatment. For both substrates, contact angles statistically higher were obtained for dentin covered with SL (P <0.05). The SL removal resulted in significant reduction of the angles (P <0.05) and no difference was found among angles produced on demineralized dentin impregnated with water or chlorhexidine (p> 0.05). Conclusion: Caries affected dentin wettability was higher than sound dentin and that characteristic was not influenced by chlorhexidine application.