Dentin matrix components extracted with phosphoric acid enhance cell proliferation and mineralization.

OBJECTIVE: Acids, such as those used in adhesive dentistry, have been shown to solubilize bioactive molecules from dentin. These dentin matrix components (DMC) may promote cell proliferation and differentiation, and ultimately contribute to dentin regeneration. The objective of this study was to evaluate the potential for varying concentrations of DMC extracted from human dentin by phosphoric acid of a range of pHs to stimulate proliferation and mineralization of two different cultured pulp cell populations. METHODS: DMC were solubilized from powdered human dentin (7 days - 4°C) by phosphoric acid of pH 1, 3, and 5 and also, EDTA. Extracts were dialyzed for 7 days against distilled water and lyophilized. Undifferentiated mouse dental pulp cells (OD-21) and cells of the odontoblast-like cell line (MDPC-23) were seeded in six-well plates (1×10(5)) and cultured for 24h in DMEM (Dulbecco's modified Eagle's medium) containing 10% (v/v) FBS (fetal bovine serum). The cells were washed with serum-free medium and then treated with different concentrations of DMC (0.01, 0.1, 1.0 and 10.0µg/ml) daily in serum free medium for 7 days. After 3, 5 (MDPC-23 only), and 7 days of treatment, cell proliferation was measured using 10vol% Alamar blue solution, which was added to each well for 1h. Cell numbers were first measured by cell counting (Trypan blue; n=5) and Alamar blue fluorescence to validate the assay, which was then used for the subsequent assessments of proliferation. Mineralization was assessed by Alizarin Red S assay after 12 days exposure to DMC (n=5). Controls were media-only (DMEM) and dexamethasone (DEX; positive control). Results were analysed by ANOVA/Tukey's (p≤0.05). RESULTS: There was a linear correlation between cell counts and Alamar blue fluorescence (R(2)>0.96 for both cell types) , verifying the validity of the Alamar blue assay for these cell types. In general, there was a dose-dependent trend for enhanced cell proliferation with higher concentration of DMC for both cell lines, especially at 10.0µg/ml. DEX exposure resulted in significantly higher mineralization, but did not affect cell proliferation. DMC exposure demonstrated significantly greater mineralization than media-only control for 10µg/ml for all extracts, and at lower concentrations for EDTA and pH 5 extracts. SIGNIFICANCE: Human dentin matrix components solubilized by acids at pH levels found in commercial dentin adhesives enhanced cell proliferation and mineralization of mouse and rat undifferentiated dental pulp cells when presented in adequate concentration.

Erosion in relation to nutrition and the environment.

When considering the erosive potential of a food or drink, a number of factors must be taken into account. pH is arguably the single most important parameter in determining the rate of erosive tissue dissolution. There is no clear-cut critical pH for erosion as there is for caries. At low pH, it is possible that other factors are sufficiently protective to prevent erosion, but equally erosion can progress in acid of a relatively high pH in the absence of mitigating factors. Calcium and phosphate concentration, in combination with pH, determine the degree of saturation with respect to tooth minerals. Solutions supersaturated with respect to enamel or dentine will not cause them to dissolve, meaning that given sufficient common ion concentrations erosion will not proceed, even if the pH is low. Interestingly, the addition of calcium is more effective than phosphate at reducing erosion in acid solutions. Today, several calcium-enriched soft drinks are on the market, and acidic products with high concentrations of calcium and phosphorus are available (such as yoghurt), which do not soften the dental hard tissues. The greater the buffering capacity of the drink or food, the longer it will take for the saliva to neutralize the acid. A higher buffer capacity of a drink or foodstuff will enhance the processes of dissolution because more release of ions from the tooth mineral is required to render the acid inactive for further demineralization. Temperature is also a significant physical factor; for a given acidic solution, erosion proceeds more rapidly the higher the temperature of that solution. In recent years, a number of interesting potentially erosion-reducing drink and food additives have been investigated.

Chlorhexidine in endodontics.

Chemical auxiliary substances (CAS) are essential for a successful disinfection and cleanness of the root canals, being used during the instrumentation and if necessary, as antimicrobial intracanal medicaments. Different CAS have been proposed and used, among which sodium hypochlorite (NaOCl), chlorhexidine (CHX), 17% EDTA, citric acid, MTAD and 37% phosphoric acid solution. CHX has been used in Endodontics as an irrigating substance or intracanal medicament, as it possesses a wide range of antimicrobial activity, substantivity (residual antimicrobial activity), lower cytotoxicity than NaOCl whilst demonstrating efficient clinical performance, lubricating properties, rheological action (present in the gel presentation, keeping the debris in suspension); it inhibits metalloproteinase, is chemically stable, does not stain cloths, it is odorless, water soluble, among other properties. CHX has been recommended as an alternative to NaOCl, especially in cases of open apex, root resorption, foramen enlargement and root perforation, due to its biocompatibility, or in cases of allergy related to bleaching solutions. The aim of this paper is to review CHX's general use in the medical field and in dentistry; its chemical structure, presentation form and storage; mechanism of action; antimicrobial activity including substantivity, effects on biofilms and endotoxins, effects on coronal and apical microbial microleakage; tissue dissolution ability; interaction with endodontic irrigants; effects on dentin bonding, metalloproteinases and collagen fibrils; its use as intracanal medicament and diffusion into the dentinal tubules; its use as disinfectant agent of obturation cones; other uses in the endodontic therapy; and possible adverse effects, cytotoxicity and genotoxicity.

Dentin matrix component solubilization by solutions of pH relevant to self-etching dental adhesives.

PURPOSE: To evaluate the potential for phosphoric acid solutions - common constituents of dental adhesive systems - of varying pH to solubilize dentin matrix components (DMCs) from human dentin. MATERIALS AND METHODS: Human dentin chips were ground under liquid nitrogen to a powder (ca 100 µm) and incubated at 4°C with agitation in phosphoric acid of pH 1, 2, 3, 4, 5, and 6 (1 g/4 ml; n = 4) for six days with solution changes each day. Estimates of daily protein release were made by UV spectrophotometry at 280 nm. Extract solutions were dialyzed for 7 days in reverse osmosis water, lyophilized, and weighed. Non-collagenous proteins (NCPs) and glycosaminoglycans (GAGs) were quantitated by dye-binding assays. 1D-PAGE for preliminary protein characterization and sandwich ELISA for presence of TGF-ß1 were performed. The results were analyzed by ANOVA and regression (α <= 0.05). RESULTS: Protein release was drastically reduced after the first few days, with the highest amounts obtained from pH 1. There was no significant difference in the quantity of DMCs solubilized by the different pH levels, but there was a significant logarithmic relation between release and pH, suggesting that greater DMC solubilization occurs with higher hydrogen ion concentrations. Dye binding assays confirmed the release of NCPs and GAGs at all pH levels. There were only subtle differences in protein bands observed between the different pH levels (1D-PAGE). Significant levels of TGF-ß1 were identified from extraction at all pHs. CONCLUSION: Acids at pH levels relevant to those used in commercial dentin adhesives are capable of solubilizing human DMCs, with release being related to hydrogen ion concentration.

Chlorhexidine in Endodontics

Chemical auxiliary substances (CAS) are essential for a successful disinfection and cleanness of the root canals, being used during the instrumentation and if necessary, as antimicrobial intracanal medicaments. Different CAS have been proposed and used, among which sodium hypochlorite (NaOCl), chlorhexidine (CHX), 17% EDTA, citric acid, MTAD and 37% phosphoric acid solution. CHX has been used in Endodontics as an irrigating substance or intracanal medicament, as it possesses a wide range of antimicrobial activity, substantivity (residual antimicrobial activity), lower cytotoxicity than NaOCl whilst demonstrating efficient clinical performance, lubricating properties, rheological action (present in the gel presentation, keeping the debris in suspension); it inhibits metalloproteinase, is chemically stable, does not stain cloths, it is odorless, water soluble, among other properties. CHX has been recommended as an alternative to NaOCl, especially in cases of open apex, root resorption, foramen enlargement and root perforation, due to its biocompatibility, or in cases of allergy related to bleaching solutions. The aim of this paper is to review CHX's general use in the medical field and in dentistry; its chemical structure, presentation form and storage; mechanism of action; antimicrobial activity including substantivity, effects on biofilms and endotoxins, effects on coronal and apical microbial microleakage; tissue dissolution ability; interaction with endodontic irrigants; effects on dentin bonding, metalloproteinases and collagen fibrils; its use as intracanal medicament and diffusion into the dentinal tubules; its use as disinfectant agent of obturation cones; other uses in the endodontic therapy; and possible adverse effects, cytotoxicity and genotoxicity.

Resumo Substâncias químicas auxiliares (SQA) são essenciais para o processo de limpeza e desinfecção dos canais radiculares, sendo utilizadas durante a instrumentação dos canais radiculares e, se necessário, como medicamentos intracanais. Diferentes SQA têm sido propostas e utilizadas, entre elas: hipoclorito de sódio (NaOCl), clorexidina (CHX), EDTA 17%, ácido cítrico, MTAD e solução de ácido fosfórico a 37%. CHX tem sido usada na endodontia como SQA ou medicação intracanal. CHX possui uma ampla gama de atividade antimicrobiana; substantividade (atividade antimicrobiana residual); menor citotoxicidade que NaOCl, demonstrando desempenho clínico eficiente; propriedades de lubrificação; ação reológica (presente na apresentação gel, mantendo os detritos em suspensão); inibe metaloproteinases; é quimicamente estável; não mancha tecidos; é inodora; solúvel em água; entre outras propriedades. CHX tem sido recomendada como uma alternativa ao NaOCl, especialmente em casos de ápice aberto, reabsorção radicular, perfuração radicular e durante a ampliação foraminal, devido à sua biocompatibilidade, ou em casos de alergia ao NaOCl. O objetivo deste trabalho é fazer uma revisão do uso da clorexidina na medicina e na odontologia; sua estrutura química; forma de apresentação e armazenamento; mecanismo de ação, atividade antimicrobiana, incluindo, substantividade, efeitos sobre biofilmes e endotoxinas; efeito sobre infiltração microbiana coronal e apical; capacidade de dissolução do tecido; interação com os irrigantes; efeitos sobre a união à dentina, metaloproteinases e fibrilas de colágeno; a sua utilização como medicamento intracanal e difusão ...

Effects of a novel fluoride-containing aluminocalciumsilicate-based tooth coating material (Nanoseal) on enamel and dentin.

PURPOSE: To investigate the effect of a fluoride-containing aluminocalciumsilicate nanoparticle glass dispersed aqueous solution (Nanoseal) on enamel and dentin, under the hypothesis that this material can form insoluble mineral deposits that confer acid resistance to the tooth structure and occlude open dentin tubules. METHODS: Labial enamel and dentin of human extracted incisors were used. Morphology of the enamel and dentin artificially demineralized with a lactic acid solution that before and/or after coated with the test material were analyzed with a wavelength-dispersive X-ray spectroscopy electron probe microanalyzer with an image observation function (SEM-EPMA). Moreover, incorporation of the calcium and silicon by enamel and dentin were also detected with SEM-EPMA. RESULTS: Application of the fluoroaluminocalciumsilicate-based tooth coating material resulted in the deposition of substances (nanoparticles) onto the enamel surface porosities and open dentin tubules on the artificial lesions. Prior coating with the test material reduced the demineralization-induced loss of enamel and dentin. Moreover, Ca and Si incorporation into superficial enamel and dentin was detected.

Mineral ions in the fluid of biofilms formed on enamel and dentine shortly after sugar challenge.

To test the effect of distinct solubilities of dentine and enamel on mineral ion concentration in the biofilm fluid during a sugar-induced pH drop, dental biofilms were formed in situ for 4 days on acrylic (control), dentine or enamel. On the 5th day, they were treated with water (control) or 20% glucose and collected 5 min later. Significantly lower pH values and higher calcium concentrations were found in the biofilm fluid after glucose exposure, without significant differences among the three substrates. During pH drop, biofilm reservoirs release calcium to the fluid, masking the differential solubility between enamel and dentine.

Evaluation of the interaction between endodontic sealers and dentin treated with different irrigant solutions.

INTRODUCTION: Dentin surface treatment with different substances might cause alterations in chemical and structural compositions of the human dentin. The aim of this study was to investigate the wettability of endodontic sealers in contact with dentin treated with 5.25% sodium hypochlorite (NaOCl) and 2% chlorhexidine (CHX) in the presence or absence of smear layer. METHODS: Thirty-six dentin slices were used. They were irrigated with NaOCl and washed with distilled water (DW) to simulate irrigation during chemomechanical preparation. After this procedure, the samples were divided into 6 groups: group 1, DW (control); group 2, NaOCl + DW; group 3, CHX + DW; group 4, ethylenediaminetetraacetic acid (EDTA) + DW; group 5, EDTA + DW + NaOCl + DW; group 6, EDTA + DW + CHX + DW. The Ramé-Hart goniometer was used to measure the contact angle between the dentin surfaces and the AH Plus and Real Seal SE sealers. Data were statistically analyzed. RESULTS: With regard to the contact angle between AH Plus and treated surfaces, the values of contact angle were lower when CHX was used, regardless of the presence or absence of smear layer. When Real Seal SE was evaluated, in the absence of smear layer, CHX presented lower values of contact angle. In the presence of smear layer, the use of NaOCl and CHX did not favor the spread of the sealer. CONCLUSIONS: The present study has revealed that smear layer removal and final flush with CHX favor the wettability of AH Plus and Real Seal SE sealers.

Influence of five home whitening gels and a remineralizing gel on the enamel and dentin ultrastructure and hardness.

PURPOSE: To investigate the influence of calcium phosphate enhanced home whitening agents on human enamel and dentin surface microhardness and ultramorphology. METHODS: Five intact molars crowns were used for ultrastructural analysis and five for microhardness test. Each resulting coronal structure was cut in slices. After measuring baseline Knoop Hardness Number (KHN) of the enamel and dentin, the slices were divided into six experimental groups and one control (n= 5). G1= 15% carbamide peroxide (CP); G2= 16% CP; G3= Ca and PO4 (remineralizing agent); G4= 16% CP with Ca and PO4; G5= 7.5% hydrogen peroxide (HP) with Ca and PO4; G6=7.5% HP with Ca. After each daily session of treatment, specimens were stored in distilled water (37 degrees C) until the next session. Products were applied for 2 weeks, according to manufacturers' instructions. Additional KHN weredetermined. RESULTS: Conventional whitening agents (G1; G2) and the gel with Ca (G6), caused KHN decrease (P< 0.05).The remineralizing and whitening agents with Ca and PO4 (G3; G4; G5) did not change KHN. A change of morphology was observed on enamel and dentin surfaces in G1; G2; G5.

Calcium phosphate-based solutions promote dentin tubule occlusions less susceptible to acid dissolution.

PURPOSE: To determine the efficacy supersaturated calcium phosphate (CaP) solutions containing fluoride (F) and zinc (Zn) ions in occluding dentin tubules with precipitates less susceptible to acid dissolution and to compare the performance of these solutions with the oxalate solutions containing calcium (Ca) or phosphate (P) ions. METHODS: Dentin sections from human molars divided into groups: Group A - control (treated with double distilled H2O), Groups A1, A2 and A3 were treated with experimental solutions supersaturated with respect to F and Zn-substituted calcium phosphates. Solutions A1 and A2 were similar in composition but differed in pH values (A1, pH 7; A2, pH 5.5). Solutions A2 and A3 were similar in pH (pH 5.5) but the A3 solution had twice the concentrations of F and Zn2+ ions compared to A2. Another group of dentin sections were treated with A3 solution, oxalate solution containing Ca (OX/Ca) and OX solution containing P (OX/P). The control and treated dentin sections were characterized using scanning electron microscopy. RESULTS: All treated dentin sections showed occluded dentin tubules; with the group A3 showing the highest percent of occluded dentin tubules. The precipitates in the dentin tubules treated with A3 remained while those treated with OX/Ca or OX/P dissolved after exposure to an acidic buffer.

In vitro evaluation of fluoride and calcium sodium phosphosilicate toothpastes, on root dentine caries lesions.

OBJECTIVES: The aim of this in vitro study was to assess the effect of toothpastes containing three different sodium fluoride concentrations and a calcium sodium phosphosilicate system, on root dentine demineralization and remineralization. METHODS: During a fourteen-day pH-cycling protocol, pre-softened bovine root dentine specimens were immersed twice daily, before and after the demineralization periods, for 2 min, in the following toothpaste slurries: (a) non-fluoridated (control), (b) 7.5% calcium sodium phosphosilicate, (c) 1450 ppm F, (d) 2800 ppm F, and (e) 5000 ppm F. Subsequently, the slabs were subjected to a fifty-hour acid resistance test. Knoop microhardness at different lesion depths was assessed in specimen cross-sections and KHN values were converted to vol.% mineral. Comparisons between the groups were performed at each lesion depth through ANOVA-based tests and furthermore, regression analysis of the derived statistic of "integrated vol.% mineral loss" was carried out. Also, lesions were evaluated qualitatively using transmission and polarized light microscopy. RESULTS: The 5000 ppm F toothpaste group, during pH-cycling, presented significantly less total vol.% mineral loss and subsequently exhibited considerably increased surface acid resistance, compared to all the other tested groups. The calcium sodium phosphosilicate toothpaste, during pH-cycling, inhibited demineralization and/or promoted remineralization of the surface layers significantly more effectively than the control group nevertheless, subsequently, the acid resistance of the calcium sodium phosphosilicate dentifrice group was similar to that of the control group. These observations were confirmed by microscopic examination of the lesions. CONCLUSIONS: Under the present experimental conditions, the 5000ppm F toothpaste, promoted remineralization and inhibited demineralization more effectively, than the other tested toothpastes.

Stabilizing sodium hypochlorite at high pH: effects on soft tissue and dentin.

INTRODUCTION: When sodium hypochlorite solutions react with tissue, their pH drops and tissue sorption decreases. We studied whether stabilizing a NaOCl solution at a high pH would increase its soft-tissue dissolution capacity and effects on the dentin matrix compared with a standard NaOCl solution of the same concentration and similar initial pH. METHODS: NaOCl solutions were prepared by mixing (1:1) a 10% stock solution with water (standard) or 2 mol/L NaOH (stabilized). Physiological saline and 1 mol/L NaOH served as the controls. Chlorine content and alkaline capacity of NaOCl solutions were determined. Standardized porcine palatal soft-tissue specimens and human root dentin bars were exposed to test and control solutions. Weight loss percentage was assessed in the soft-tissue dissolution assay. Three-point bending tests were performed on the root dentin bars to determine the modulus of elasticity and flexural strength. Values between groups were compared using one-way analysis of variance with the Bonferroni correction for multiple testing (α < .05). RESULTS: Both solutions contained 5% NaOCl. One milliliter of the standard and the stabilized solution consumed 4.0 mL and 13.7 mL of a 0.1-mol/L HCl solution before they reached a pH level of 7.5, respectively. The stabilized NaOCl dissolved significantly more soft tissue than the standard solution, and the pH remained high. It also caused a higher loss in elastic modulus and flexure strength (P < .05) than the control solutions, whereas the standard solution did not. CONCLUSIONS: NaOH-stabilized NaOCl solutions have a higher alkaline capacity and are thus more proteolytic than standard counterparts.

Smear layer dissolution by peracetic acid of low concentration.

AIM: To test the effect of a noncaustic concentration of peracetic acid (PAA) in a standardized smear layer model. METHODOLOGY: The smear layer dissolution kinetics of 0.5% PAA on human dentine were compared to those of 2.25% PAA and 17% ethylenediaminetetraacetic acid (EDTA) solutions. Coronal dentine discs were prepared from six human maxillary molars. A standardized smear layer was produced on the pulpal side of each disc. The smear layer-covered surface was divided into three similar areas and then exposed to one of the three solutions tested. Co-site image sequences (around 40, 500 ×) of the specific areas were obtained after four cumulative demineralisation times (15, 30, 60 and 180 s). An image processing and analysis sequence measured sets of images, providing data of area fraction (AF, dentine-free area in % of total analysis area). A general linear model for repeated measures was used to verify the influence of time and solution type over the change in AF from baseline (ΔAF). RESULTS: Overall, EDTA and 2.25% PAA produced higher ΔAF values than the 0.5% PAA solution (P < 0.05). No significant difference was observed in ΔAF between 15 s and 30 s (P > 0.05). After 60 s of etching, all tested solutions produced similar ΔAF (P > 0.05), whereas at 180 s, ΔAF of both EDTA and 2.25% PAA continued to increase (P > 0.05). CONCLUSIONS: After 60 s of contact, the 0.5% PAA solution dissolved smear layer as well as 2.25% PAA and 17% EDTA.

The impact of root dentine conditioning on sealing ability and push-out bond strength of an epoxy resin root canal sealer.

AIM: To investigate the impact of dentine conditioning on sealing ability and dentine bond strength of an epoxy resin sealer. METHODOLOGY: Root canals in 90 single-rooted teeth were instrumented using a rotary Ni-Ti system. Fifty canals were irrigated with water during instrumentation, 40 with 3% NaOCl. A final flush was performed in the water-irrigated specimens with water (negative control), 3% NaOCl, 17% EDTA, 7% maleic acid (MA) or 2% chlorhexidine. The hypochlorite irrigated specimens received a final flush with a decalcifying agent (EDTA or MA) and then 3% NaOCl or 3% NaOCl and then the decalcifying agent (n = 10, each). Canals were all filled with AH Plus. Fluid transport was measured on day 3 and 30. Roots were then sectioned, and push-out tests were performed in coronal, middle and apical root thirds. Results were analysed using analysis of variance (anova) with Bonferroni's adjustment. Spearman's rank correlation was computed between fluid transport and push-out bond strength. RESULTS: Leakage decreased over time (P < 0.05). Push-out bond strength was highest in coronal and lowest in apical root thirds (P < 0.05). Irrigating protocols with final application of a decalcifying agent greatly decreased the leakage and increased push-out bond strength values, in contrast to groups where NaOCl was applied last (P < 0.05), wherein the effect of the decalcifying agent was abolished. Chlorhexidine had no impact on the outcomes. Fluid transport and push-out bond strength correlated strongly (ρ = -0.83). CONCLUSIONS: AH Plus appears to bond to the organic phase of dentine. This bond influences its sealing ability.

The inhibitory effects of quaternary ammonium methacrylates on soluble and matrix-bound MMPs.

Matrix metalloproteinases (MMPs) bound to dentin contribute to the progressive degradation of collagen fibrils in hybrid layers created by dentin adhesives. This study evaluated the MMP-inhibiting potential of quaternary ammonium methacrylates (QAMs), with soluble rhMMP-9 and a matrix-bound endogenous MMP model. Six different QAMs were initially screened by a rhMMP-9 colorimetric assay. For the matrix-bound endogenous MMPs, we aged demineralized dentin beams for 30 days in calcium- and zinc-containing media (CM; control), chlorhexidine, or QAMs in CM to determine the changes in dry mass loss and solubilization of collagen peptides against baseline levels. The inhibitory effects of QAMs on soluble rhMMP-9 varied between 34 and 100%. Beams incubated in CM showed a 29% decrease in dry mass (p < 0.05), whereas beams incubated with QAMs showed only 0.2%-6% loss of dry mass. Significantly more solubilized collagen was detected from beams incubated in CM (p < 0.05). It is concluded that QAMs exhibited dentin MMP inhibition comparable with that of chlorhexidine, but required higher concentrations.

The requirement of zinc and calcium ions for functional MMP activity in demineralized dentin matrices.

UNLABELLED: The progressive degradation of resin-dentin bonds is due, in part, to the slow degradation of collagen fibrils in the hybrid layer by endogenous matrix metalloproteinases (MMPs) of the dentin matrix. In in vitro durability studies, the storage medium composition might be important because the optimum activity of MMPs requires both zinc and calcium. OBJECTIVE: This study evaluated the effect of different storage media on changes in matrix stiffness, loss of dry weight or solubilization of collagen from demineralized dentin beams incubated in vitro for up to 60 days. METHODS: Dentin beams (1mm×2mm×6mm) were completely demineralized in 10% phosphoric acid. After baseline measurements of dry mass and elastic modulus (E) (3-point bending, 15% strain) the beams were divided into 5 groups (n=11/group) and incubated at 37°C in either media containing both zinc and calcium designated as complete medium (CM), calcium-free medium, zinc-free medium, a doubled-zinc medium or water. Beams were retested at 3, 7, 14, 30, and 60 days of incubation. The incubation media was hydrolyzed with HCl for the quantitation of hydroxyproline (HOP) as an index of solubilization of collagen by MMPs. Data were analyzed using repeated measures of ANOVA. RESULTS: Both the storage medium and the storage time showed significant effects on E, mass loss and HOP release (p<0.05). The incubation in CM resulted in relatively rapid and significant (p<0.05) decreases in stiffness, and increasing amounts of mass loss. The HOP content of the experimental media also increased with incubation time but was significantly lower (p<0.05) than in the control CM medium, the recommended storage medium. CONCLUSIONS: The storage solutions used to age resin-dentin bonds should be buffered solutions that contain both calcium and zinc. The common use of water as an aging medium may underestimate the hydrolytic activity of endogenous dentin MMPs.

Development of an acid challenge-based in vitro dentin disc occlusion model.

OBJECTIVE: The objective of this study was to evaluate the utility of a novel acid challenge-based dentin disc occlusion model, and to compare the occluding effect and acid resistance exhibited by currently marketed occlusion dentifrices in vitro. METHODS: Ninety-six bovine dentin discs were polished and etched in citric acid (6% w/w) for two minutes to provide a smooth dentin surface with patent tubules. The discs were divided into three treatment groups. Each treatment group was brushed (Oral-B Vitality Precision Clean/EB 17 FlexiSoft head) twice a day, for up to four days, with either a strontium acetate dentifrice (Sensodyne Rapid Relief), an arginine-based dentifrice (Colgate Sensitive Pro-Relief), or water. Prior to and between treatments, the dentin samples were stored in human saliva. On days 3 and 4, following dentifrice treatment and incubation in saliva (60 minutes), the samples were subjected to a grapefruit juice challenge. Eight samples from each treatment group were removed from the study on each day and analyzed by scanning electron microscopy (SEM). The SEM images were graded according to a categorical occlusion scale, and the data were analyzed by ANOVA. RESULTS: The strontium acetate dentifrice occluded dentin tubules significantly better than the negative control (water) on days 1 through 4 (day 4 p < or = 0.0001). The marketed occlusion-based dentifrices demonstrated various degrees of dentin tubule occlusion over the four days. The strontium acetate dentifrice demonstrated significantly better occlusion than a currently marketed arginine-based occlusion dentifrice on day 1 (p = 0.0337), day 2 (p = 0.0021 ), and day 4 (p < or = 0.0001). CONCLUSION: An in vitro model has been developed that can differentiate the dentin tubular occlusive effects of currently marketed occlusion dentifrices. Surface analysis reveals that the occluding deposits vary according to product, and that some are more susceptible to acid mediated dissolution.

A minimally destructive technique for removing the smear layer from dentine surfaces.

OBJECTIVES: To develop a minimally destructive technique for removing the smear layer produced by cutting and polishing specimens of dentine prepared for use in experimental studies, e.g. on occlusion of dentinal tubules by oral health products. The aim was to avoid the damage caused by conventional techniques utilising short exposures to solutions with very low pH. METHODS: Two acetate buffers, pH 5.5, containing different concentrations of calcium and phosphate, with -log(ion activity product with respect to hydroxyapatite) (pI(HA)) of 55 or 56, were tested on slices of dentine using scanning electron microscopy (SEM). RESULTS: A solution which, from previous work, was slightly undersaturated with respect to dentine mineral, with a pI(HA) of 56, was found to remove smear layers produced by cutting and/or polishing after 15 min. However, to reliably remove debris occluding the tubules an exposure time of 2h, followed by brief ultrasonication, was necessary. After 2h treatment with this buffer, only a small amount of demineralization of the surface was detectable by SEM, while calcium and phosphorus were detectable by X-ray dispersive spectroscopy. CONCLUSION: It is possible to remove smear layers, and to open dentinal tubules, by a reasonably short exposure to an acidic buffer which is undersaturated with respect to dentine mineral.

A study of the relation between erosion and microhardness of root canal dentin.

OBJECTIVE: The aim of this study was to find a relation between erosion and microhardness of root canal dentin after irrigation with different canal irrigants. STUDY DESIGN: Seventy-two single-canal human premolar teeth were selected and enlarged by rotary Protaper files. The middle part of each root was transversely sectioned to a 4-mm slice. The initial microhardness values of intact specimens were measured at depths of 100 microm and 500 microm from the pulp-dentin interface using a Vickers microhardness tester. the specimens were divided into 6 groups of 12 specimens and were treated as follows: 1: 2.6% NaOCl, 2: 17% EDTA (5 minutes) then 2.6% NaOCl (5 minutes), 3: 17% EDTA (1 minute) then 2.6% NaOCl (1 minute), 4: MTAD (5 minutes), 5: 2% Chlorhexidine (5 minutes), and 6: saline (control), respectively. Posttreatment microhardness values were obtained in the same manner as the initial ones. Afterwards, the specimens were prepared for scanning electron microscopy analysis. The amount of dentin erosion was examined. RESULTS: Group 2 showed the most erosive effect on dentin (P < .0001) along with the least decrease of dentin microhardness at depth of 100 microm, whereas MTAD showed the most reduction of dentin microhardness and less erosive effect on dentin. CONCLUSION: It can be concluded that erosion is not the main factor in decreasing the dentin microhardness, whereas the amount of irrigant penetration might be the main cause.

Impact of irrigant sequence on mechanical properties of human root dentin.

To get the root canal system free of organic debris and the smear layer, it has been recommended to irrigate with a NaOCl solution during instrumentation, followed by a rinse with a chelating agent such as ethylenediaminetetraacetic acid (EDTA) and a final irrigation with NaOCl. However, both hypochlorite and EDTA weaken dentin through dissolution of its organic and inorganic components, respectively. EDTA exposes the organic dentin matrix, which could then be attacked more easily by hypochlorite. It was the aim of this study to assess the impact of different irrigation sequences of NaOCl (2.5% w/v; total exposure time, 24 minutes) and EDTA (17%; 3 minutes) on the elastic modulus and flexure strength of standardized human root dentin bars (n = 11 per group). Exposures to solely EDTA (3 minutes), NaOCl (24 minutes), and water were used as control treatments. Specimens were subjected to 3-point bending tests; modulus of elasticity and flexure strength values were compared between groups with one-way analysis of variance followed by Fisher probable least-squares difference test. The alpha-type error was set at .05. The 24-minute exposure to the hypochlorite solution caused a significant drop in flexure strength compared with water- or EDTA-treated controls (P < .05), whereas the elastic modulus remained unaffected. In contrast, the short exposure to EDTA as is clinically recommended did not affect the mechanical dentin parameters under investigation, regardless of the irrigant sequence that was used.