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Aim To evaluate the remineralization ability of two endodontic sealer cements. Methodology Mid-coronal dentine surfaces were subjected to: (i) 37% phosphoric acid ( PA) or (ii) 0.5 mol L−1 ethylenediaminetetraacetic acid ( EDTA) conditioning prior to the application of two experimental hydroxyapatite-based cements, containing sodium hydroxide (calcypatite) or zinc oxide oxiapatite respectively. Samples were stored in simulated body fluid for 24 h or 21 days. Remineralization of the dentine surfaces were studied by Raman spectroscopy (mapping with K-means cluster and hierarchical cluster analysis) was undertaken. Nanoroughness and collagen fibril width measurements were performed with an atomic force microscopy. ANOVA and Student-Newman-Keuls test were performed (α=0.05). Results Phosphoric acid+oxiapatite promoted both the highest dentine mineralization ( P < 0.05) and crystallographic maturity at the dentine surface. Noncrystalline amorphous-like apatites were also formed. Dentine treated with PA+calcypatite attained the roughest surface ( P < 0.05) with minimal fibril width ( P < 0.05). Cross-linking of collagen only became greater in the group PA+oxiapatite after 21 days. The maximum relative mineral concentration and structure of collagen linked to the amide I and ratio amide III/ AGEs was obtained after using PA+calcypatite at 21-days time-point ( P < 0.05). EDTA produced a lower stoichiometric hydroxyapatite ( P < 0.05) with decreased maturity, at the expense of carbonate band widening, although it favoured the nucleation of carbonated calcium phosphate. Conclusions Dentine surfaces treated with PA+oxiapatite attained the highest dentine remineralization with both crystalline-stoichiometric and amorphous apatites, at 21 days. EDTA conditioning facilitated amorphous-bulk mineral precipitation. The amorphization was more intense after using oxiapatite and provided an ion-rich environment favouring in situ dentine remineralization. [ABSTRACT FROM AUTHOR]

To obtain more durable adhesion to dentin, and to protect collagen fibrils of the dentin matrix from degradation, calcium- and phosphate-releasing particles have been incorporated into the dental adhesive procedure. The aim of the present study was to incorporate zinc-loaded polymeric nanocarriers into a dental adhesive system to facilitate inhibition of matrix metalloproteinases (MMPs)-mediated collagen degradation and to provide calcium ions for mineral deposition within the resin-dentin bonded interface. PolymP-nActive nanoparticles (nanoMyP) were zinc-loaded through 30-minute ZnCl2 immersion and tested for bioactivity by means of 7 days’ immersion in simulated body fluid solution (the Kokubo test). Zinc-loading and calcium phosphate depositions were examined by scanning and transmission electron microscopy, elemental analysis, and x-ray diffraction. Nanoparticles in ethanol solution infiltrated into phosphoric-acid-etched human dentin and Single Bond (3M/ESPE) were applied to determine whether the nanoparticles interfered with bonding. Debonded sticks were analyzed by scanning electron microscopy. A metalloproteinase collagen degradation assay was also performed in resin-infiltrated dentin with and without nanoparticles, measuring C-terminal telopeptide of type I collagen (ICTP) concentration in supernatants, after 4 wk of immersion in artificial saliva. Numerical data were analyzed by analysis of variance (ANOVA) and Student-Newman-Keuls multiple comparisons tests (p < .05). Nanoparticles were effectively zinc-loaded and were shown to have a chelating effect, retaining calcium regardless of zinc incorporation. Nanoparticles failed to infiltrate demineralized intertubular dentin and remained on top of the hybrid layer, without altering bond strength. Calcium and phosphorus were found covering nanoparticles at the hybrid layer, after 24 h. Nanoparticle application in etched dentin also reduced MMP-mediated collagen degradation. Tested nanoparticles may be incorporated into dental adhesive systems to provide the appropriate environment in which dentin MMP collagen degradation is inhibited and mineral growth can occur. [ABSTRACT FROM PUBLISHER]

The aim of this study was to ascertain whether zinc may improve the repair ability of demineralized dentin. Dentin disks were demineralized by phosphoric acid during 15 s and immersed in artificial saliva, remineralizing solution, a zinc chloride solution and a zinc oxide solution. Dentin specimens were analyzed after 24 h and 1 month of storage. Surface morphology was assessed by atomic force and scanning electron microscopy, mechanical properties were analyzed by nanohardness testing in a TriboIndenter, and chemical changes at the surfaces were determined by X-ray diffraction, Raman and energy-dispersive elemental analyses. After phosphoric acid application, dentin was only partially demineralized. Demineralized dentin was remineralized after 24 h of storage in any of the tested solutions (nanohardness increased and hydroxylapatite formation was detected by Raman). Remineralization was maintained up to 1 month in dentin stored in remineralizing solution, zinc chloride and zinc oxide. Zinc and phosphate were important for hydroxylapatite homeostasis. Scholzite formation was encountered in dentin stored in zinc-containing solutions. Zinc might allow to reach the balance between dentin demineralization and remineralization processes. © 2014 S. Karger AG, Basel [ABSTRACT FROM AUTHOR]

Background: Dentine matrix metalloproteinases (MMPs) play an important role in the dentine caries process. Aims: To determine if MMP-mediated collagen degradation of acid-demineralized dentine may be inhibited by zinc or zinc chelators. Methods: Human dentine specimens were demineralized by phosphoric acid (PA), ethylenediaminetetraacetic acid (EDTA), Clearfil SE Bond primer (SE), or Xeno V (XE) and stored in artificial saliva. Chlorhexidine digluconate (CHX), doxycycline, EDTA, or ZnCl2 was added. C-terminal telopeptide determinations (ICTP) were performed by radioimmunoassay after 24 h and 4 weeks. Results: Collagen degradation was prominent in PA-demineralized (ICTP values from 74.01 μg/l at 24 h to 202.46 μg/l after 4 weeks) and EDTA-demineralized dentine (ICTP values from 83.93 μg/l at 24 h to 158.82 μg/l after 4 weeks) stored in artificial saliva. Doxycycline fully blocked proteolysis. CHX and EDTA reduced collagen degradation only at 24 h. Zinc in excess strongly inhibited hydrolysis of collagen in all tested groups (ICTP values were: PA, 13.56 μg/l; EDTA, 11.21 μg/l; SE, 1.52 μg/l, and XE, 2.37 μg/l) and its effect was maintained for up to 4 weeks, except for EDTA-treated dentine (ICTP values were: PA, 40.76 μg/l; EDTA, 79.15 μg/l; SE, 5.29 μg/l, and XE, 6.38 μg/l). Conclusion: EDTA and CHX exerted time-limited MMP inhibition, and excess zinc served as an effective inhibitor of MMP-mediated collagen degradation in strong or mildly demineralized dentine. MMP degradation of collagen was reduced in resin-infiltrated dentine; the presence of excess zinc chloride exerted an additional protective effect. Copyright © 2012 S. Karger AG, Basel [ABSTRACT FROM AUTHOR]

This study investigated the microtensile bond strength (μTBS) of a one-step self-etching adhesive to human dentin and bovine enamel following different bonding treatments. Occlusal portions of human molars and labial surfaces of bovine incisors were ground flat to provide uniform dentin and enamel surfaces, respectively. Futurabond was used following five different protocols: 1) according to the manufacturer's directions, 2) acid etched with 36% phosphoric acid (H3PO4) for 15 seconds, 3) 10% sodium hypochlorite (NaOCl) treated for two minutes after H3PO4-etching, 4) doubling the application time of the adhesive and 5) doubling the number of adhesive coats. Composite build-ups (6 mm in height) were constructed incrementally with Arabesk resin composite. The specimens were stored in 100% humidity for 24 hours at 37°C and sectioned into beams of 1.0 mm² cross-sectional area. Each beam was tested in tension in an Instron machine at 0.5 mm/minute, and mean μTBS data (MPa) were analyzed by one-way ANOVA and post-hoc multiple comparisons tests (α=0.05). Doubling the application time of Futurabond attained the highest μTBS to dentin; whereas, no differences among all bonding application parameters evaluated could be detected when the adhesive was applied to enamel. [ABSTRACT FROM AUTHOR]

The existence of unprotected collagen fibrils within the hybrid layer compromises the longevity of restorations. This phenomenon may be avoided if solutions other than strong acids are used for dentin demineralization. The hypothesis to be tested is that bond durability may be improved by EDTA demineralization. Dentin surfaces (human and bovine) were bonded: (1) after phosphoricacid- etching, and after EDTA demineralization with (2) a total-etch adhesive and (3) a selfetching adhesive. After the teeth were sectioned into beams, half of the specimens were immersed in NaOCl, while the other half was immersed in water. Beams were tested to failure in tension. ANOVA and multiple-comparisons tests were used (P < 0.05). No differences in bond strength were found among the 3 bonding procedures, although bonds made to human molars were 43- 61% higher than those to bovine incisors. After NaOCl immersion, only specimens subjected to EDTA demineralization maintained the initial bond strength. We conclude that the collagen network is better-preserved after EDTA demineralization. [ABSTRACT FROM AUTHOR]

The role of the collagen fibers in dentin adhesion has not clearly been established. Therefore, this laboratory study evaluated the microleakage at resin-dentin and resin-enamel interfaces of Class V composite restorations after etching enamel and dentin with phosphoric acid (H3PO4) or after etching with H3PO4 followed by deproteinization with 5% sodium hypochlorite (NaOCl) to prevent the formation of a hybrid layer. Ten extracted human molars were used to prepare standardized Class V cavities on both buccal and lingual surfaces. The teeth were randomly divided in two H3PO4 for 15 seconds; b) Class V cavities that were etched with H3PO4 for 15 seconds followed by collagen removal with 5% NaOCl for two minutes. The cavities were restored using the Prime & Bond 2.1 bonding system and TPH resin composite. The specimens were stored in water for 24 hours at 37°C and thermocycled 500 times between water baths kept at 5°C and 55°C. After thermocycling, specimens were immersed in a 0.5% aqueous solution of basic fuchsin for 24 hours. Three longitudinal sections of each restoration were obtained and examined with a stereomicroscope for qualitative evaluation of microleakage. The data were statistically analyzed by Mann-Whitney U and Wilcoxon matched pairs signed ranked tests. Extra specimens were analyzed with the scanning electron microscope (SEM). Occlusal margins (enamel margins) resulted in statistical lower degree of leakage-than gingival margins (dentin/cementum margins) in both treatment groups. For each type of margin, there were no statistically significant differences between the etched and the etched and deproteinized groups. Under the SEM, occlusal surfaces showed no detachment between enamel and dentin, while dentin/cementum resulted in gap formation. [ABSTRACT FROM AUTHOR]

Objectives: Dentin matrix metalloproteinases are implicated in the pathogenesis of caries and contribute to collagen degradation in resin-dentin interfaces. The objective was to determine if collagen degradation may be modulated by an excess of zinc or zinc chelators.Methods: Mineralized and phosphoric acid demineralized human dentin specimens were tested. Chlorhexidine digluconate, doxycycline or ZnCl₂ were added to the media. In half of the groups, active exogenous metalloproteinase-2 was incorporated into the solution. C-terminal telopeptide determinations (radioimmunoassay) were performed after 24 h, 1 and 3 weeks.Results: Collagen degradation was prominent in demineralized dentin. Doxycycline fully blocked dentin proteolysis. Chlorhexidine digluconate reduced the degradation at the 24-h period. Zinc in excess strongly inhibits hydrolysis of collagen and its effect was maintained for 3 weeks.Conclusions: Zinc in excess reduces MMP-mediated collagen degradation. The hypothesis that binding of zinc to collagen results in protection of sensitive cleavage sites of metalloproteinases requires further validation. [ABSTRACT FROM AUTHOR]

Abstract: Objective: To examine the differential expression of matrix metalloproteinase-2 (MMP-2) in human coronal and radicular sound and carious dentine using combined trichrome staining technique and immunofluorescence approach. Methods: Freshly extracted human premolars were fixed with formaldehyde, demineralised with 10% EDTA (pH 7.4), dehydrated and sectioned for light and immunofluorescence microscopy. Half of the sections were stained with Masson''s trichrome and examined with light microscopy to identify regions in the coronal and radicular parts of the teeth that contained sound, caries-affected and caries-infected dentine. The rest of the sections were hybridized with anti-mouse MMP-2 primary antibody and FITC-conjugated secondary antibody. Immunofluorescence of the FITC that was indicative of the distribution of the MMP-2 in coronal and radicular dentine was analysed by fluorescence light microscopy. Results: Trichrome staining revealed a green zone of unaffected sound dentin, red irregular regions of caries-infected dentine and pink regions of caries-affected dentine. Immunofluorescence signals that were indicative of MMP expression were the lowest in sound dentine and most intense in the caries-infected dentine. Caries-affected dentine showed intermediate immunoreactivity. The variations in the intensities of immunofluorescence corresponded well with the distribution of caries-infected and caries-affected dentine in the trichrome-stained sections, for both coronal and radicular dentine. Conclusion: Caries stimulates MMP-2 expression, resulting in the differential expression of this protease in sound, caries-affected and caries-infected dentine. The more intense MMP-2 expression in caries-affected dentine compared with sound dentine may imply more rapid hybrid layer degradation when caries-affected dentine is employed as the substrate for bonded restorations. [Copyright &y& Elsevier]