Optimization of calcium concentration of saliva with phosphoryl oligosaccharides of calcium (POs-Ca) for enamel remineralization in vitro

ObjectivePhosphoryl oligosaccharides of calcium (POs-Ca) are highly soluble calcium source made from potato starch. The aim of this study was to investigate the optimal concentrations of POs-Ca for the remineralization of subsurface enamel lesions in vitro.DesignDemineralized bovine enamel slabs (n = 5) were remineralized in vitro for 24 h at 37 °C with artificial saliva (AS) containing 0–0.74% POs-Ca to adjust the Ca/P ratio to 0.4–3.0, then sectioned and analysed by transversal microradiography (TMR). The data were analysed by Scheffe's post hoc test. The Ca/P ratio with most remineralization was used to investigate the effect of calcium on enamel remineralization (n = 11). The demineralized slabs were treated with AS with calcium-chloride- (CaCl₂-) or POs-Ca with an identical calcium content, and sectioned for TMR and wide-angle X-ray diffraction (WAXRD) analyses to evaluate the local changes in hydroxyapatite (HAp) crystal content. The data were analysed using the Mann–Whitney U-test.ResultsThe highest mineral recovery rate resulted from addition of POs-Ca to adjust the Ca/P to 1.67. At this ratio, the mineral recovery rate for AS containing POs-Ca (24.2 ± 7.4%) was significantly higher than that for AS containing CaCl₂ (12.5 ± 11.3%) (mean ± SD, p < 0.05). The recovery rate of HAp crystallites for AS containing POs-Ca (35.7 ± 10.9%) was also significantly higher than that for AS containing CaCl₂ (23.1 ± 13.5%) (p < 0.05). The restored crystallites were oriented in the same directions as in sound enamel.ConclusionsPOs-Ca effectively enhances enamel remineralization with ordered HAp at a Ca/P ratio of 1.67.     

Effect of iron on enamel demineralization and remineralization in vitro

ObjectiveTo evaluate the effect of ferrous sulphate on enamel demineralization and remineralization, using pH-cycling models.DesignFifty blocks were selected by their initial surface hardness and subjected to a pH-cycling demineralization process. Artificially demineralized lesions were produced in 60 blocks; out of these blocks, the surface hardness of 50 blocks and the cross-sectional hardness of 10 blocks were determined. The 50 blocks were then subjected to a remineralization pH-cycling process. Treatments were carried out using ferrous sulphate solutions of different concentrations (0.333, 0.840, 18.0, and 70.0 μg Fe/mL) and a control group (deionized water). The final surface hardness (SH₂) was determined, and the integrated subsurface hardness (ΔKHN) was calculated. The enamel blocks were analysed for fluoride, calcium, phosphorus, and iron. The obtained data were distributed heterogeneously and were analysed using the Kruskal–Wallis test (p < 0.05).ResultsIn demineralization pH cycling, the group treated with the 18.0 μg Fe/mL solution had higher secondary surface hardness and lower integrated subsurface hardness (ΔKHN) than the other groups. In remineralization pH cycling, the control group showed the lowest value of ΔKHN. A decline in Ca and P concentration was observed when the Fe concentration increased (p < 0.05). There was no significant difference in the F concentration (p > 0.05) and an increase in Fe concentration (p < 0.05) in the enamel was observed when the Fe concentration increased in both the demineralization and remineralization experiments.ConclusionThe results suggest that iron reduces demineralization but does not allow remineralization to occur.     

Evaluation of the effect of PVA tape supplemented with 2.26% fluoride on enamel demineralization using microhardness assessment and scanning electron microscopy: In vitro study

ObjectiveThis in vitro study was designed to evaluate the effectiveness of experimental 2.26% fluoride-polyvinyl alcohol (F-PVA) tape in inhibition of enamel demineralization using enamel surface microhardness (SMH) analysis and scanning electron microscopy (SEM) examination.DesignEnamel specimens (n = 60) prepared from bovine incisor teeth with microhardness ranging from 260 to 370 Knoop hardness number (KHN) were pooled and randomly assigned to four groups: control group, F-PVA tape group, F-varnish group, and CPP-ACFP group. After topical application of agents in each group, pH-cycling was processed. Then, SMH was measured and the percentage loss of surface microhardness (%SML) was calculated. For the SEM examination, five sample specimens in each group were treated and the morphologic character was evaluated.ResultsAfter pH-cycling, the SMH values of the enamel specimens of F-PVA tape and F-varnish group were significantly higher than that of CPP-ACFP group, there was no significant difference between F-PVA tape and F-varnish group. With SEM examination, enamel surface in the F-PVA tape group showed mild irregularity in general and numerous spherical and ovoid crystals formed on the enamel surface were also observed. The density of crystals was higher than that of both control group and CPP-ACFP group.ConclusionF-PVA tape is effective in inhibition of enamel demineralization. The inhibition potential of enamel demineralization of F-PVA tape is comparable with that of fluoride varnish and greater than that of CPP-ACFP. With the significant advantages of excellent physical properties, ease of application, and improved safety, F-PVA tape can be used not only for professional treatment, but also in a home-use delivery system.     

The role of grape seed extract in the remineralization of demineralized dentine: Micromorphological and physical analyses

ObjectivesGrape seed extract (GSE) is known to have a positive effect on the demineralization and/or remineralization of artificial root caries lesions. The present study aimed to investigate whether biomodification of caries-like acid-etched demineralized dentine, using proanthocyanidins-rich GSE, would promote its remineralization potential.DesignsDentine specimens were acid-etched for 30 s, then biomodified using proanthocyanidin-based preconditioners (at different concentrations and pH values) for 2 min, followed by a 15-day artificial remineralization regimen. They were subsequently subjected to microhardness measurements, micromorphological evaluation and X-ray diffraction analyses. Stability of the preconditioners was also analyzed, spectrophotometrically.ResultsA concentration-dependent increase was observed in the microhardness of the specimens that were biomodified using GSE preconditioners, without pH adjustment. Field emission scanning electron microscopy revealed greater mineral deposition on their surfaces, which was further identified mainly as hydroxylapatite. The absorbances of preconditioner dilutions at pH 7.4 and pH 10.0 decreased at the two typical polyphenol bands.ConclusionsTransient GSE biomodification promoted remineralization on the surface of demineralized dentine, and this process was influenced by the concentration and pH value of the preconditioner. GSE preconditioner at a concentration of 15%, without pH adjustment, presented with the best results, and this may be attributed to its high polyphenolic content.     

Experimental composites containing quaternary ammonium methacrylates reduce demineralization at enamel-restoration margins after cariogenic challenge

ObjectiveThis study evaluated the influence of experimental composites containing quaternary ammonium monomers (QAM) at different concentrations and alkyl chains on demineralization at enamel-composite margins after cariogenic challenge.MethodsStandardized 4 × 4 mm cavities were cut into 35 bovine enamel blocks, which were randomly divided into seven groups (n = 5) and restored with the following experimental composites and commercial materials: (G12.5) – 5% dimethylaminododecyl methacrylate (DMADDM) with a 12-carbon alkyl chain (G12.10) – 10% DMADDM, (G16.5) – 5% dimethylaminohexadecyl methacrylate (DMAHDM) with a 16-carbon alkyl chain (G16.10) – 10% DMAHDM, (CG) – control group (without QAM), (GZ250) – commercial composite (Filtek Z250^®), and (GIC) – glass ionomer cement (Maxxion R^®). After restorative procedures, initial microhardness was measured and experimental composites were subjected to Streptococcus mutans biofilm formation for 48 h. After cariogenic challenge, the samples were washed and microhardness was reassessed. A 3D non-contact profilometer was used to determine surface roughness and enamel demineralization was assessed by micro-CT. Microhardness results were analyzed by the Kruskal–Wallis and Mann-Whitney tests and micro-CT results were analyzed by Tukey’s HSD test (95% confidence interval).ResultsNone of the materials could prevent mineral loss at the enamel-restoration margins. The addition of 10% DMAHDM yielded the lowest, albeit statistically significant, mineral loss (p < 0.05). 3D non-contact profilometry showed enamel surface roughness modification after biofilm exposure. The CG had the highest roughness values. Micro-CT analysis revealed mineral loss, except for GIC.SignificanceThe addition of 10% QAM with a 16-carbon chain in experimental composites reduced mineral loss at the enamel-restoration margins after cariogenic challenge.     

Effect of bovine lactoferrin on enamel demineralization and acid fermentation by Streptococcus mutans

The purpose of this study was to evaluate the effects of bovine lactoferrin on acid fermentation and enamel demineralization using Streptococcus mutans in a culture system and an artificial mouth model system. The antibacterial activity of bovine lactoferrin (bLF) against S. mutans was analyzed by a radial diffusion assay. In the culture system, the effect of bLF on the synthesis and adherence of water insoluble glucan (WIG) and the adherence of S. mutans to a glass surface was examined by a batch culture. In the artificial mouth model system, cell suspension of S. mutans, heart infusion broth supplemented with sucrose, and PBS or lactoferrin solution were supplied separately and constantly for 21 hours. The following parameters were determined for evaluation: the amount of artificial biofilm, the changes in pH underneath the biofilm; and the changes in enamel microhardness measured by a Vicker's hardness tester. The antibacterial activity of bLF against S. mutans was observed. The amounts of bacterial cells in the total adherent fractions were inhibited by bLF in a dose dependent manner. The amounts of WIG in a firm-adherent fraction were significantly inhibited by 0.1–1.0% bLF. The changes in microhardness on enamel slabs in the bLF group (2.4 ± 0.8) showed significantly less hardness reduction than those in the control group (22.3 ± 2.5) (P < 0.001). The artificial biofilm accumulation was not reduced by bLF. The results of this study suggest that bLF might have inhibitory effects against acid fermentation and demineralization of enamel by S. mutans.     

In vitro study of the effects of fluoride-releasing dental materials on remineralization in an enamel erosion model

ObjectivesThis study was conducted to compare the remineralization effects of five regimens on the loss of fluorescence intensity, surface microhardness, roughness and microstructure of bovine enamel after remineralization. We hope that these results can provide some basis for the clinical application of these materials.MethodsOne hundred bovine incisors were prepared and divided into the following five groups, which were treated with distinct dental materials: (1) Clinpro? XT varnish (CV), (2) F-varnish (FV), (3) Tooth Mousse (TM), (4) Fuji III LC^® light-cured glass ionomer pit and fissure sealant (FJ) and (5) Base Cement^® glass polyalkenoate cement (BC). Subsequently, they were detected using four different methods: quantitative light-induced fluorescence, microhardness, surface 3D topography and scanning electron microscopy (SEM).ResultsThe loss of fluorescence intensity of CV, BC and FJ groups showed significant decreases after remineralization (p < 0.05). The microhardness values of the BC group were significantly higher than those of the other groups (p < 0.05) after 6 weeks of remineralization. The CV group's surface roughness was significantly lower than those of the other groups after 6 weeks of remineralization (p < 0.05). Regarding microstructure values, the FV group showed many round particles deposited in the bovine enamel after remineralization. However, the other four groups mainly showed needle-like crystals.ConclusionsGlass ionomer cement (GIC)-based dental materials can promote more remineralization of the artificial enamel lesions than can NaF-based dental materials. Resin-modified GIC materials (e.g., CV and FJ) have the potential for more controlled and sustained release of remineralized agents. The effect of TM requires further study.     

Influence of dentin pretreatment with 2.5% titanium tetrafluoride on inhibiting caries at the tooth-restoration interface in situ

ObjectiveInvestigate the effects of dentin pretreatment with 2.5% titanium tetrafluoride (TiF₄) aqueous solution followed by two-step self-etching (CLE/Clearfil SE Bond) and one-step self-etching adhesive systems (SBU/Single Bond Universal) on carious lesion inhibition at the tooth-restoration interface using an in situ model.DesignSixty-four cavities at the enamel-dentin junction of dental fragments were randomly distributed according to groups (n = 16): 1) TiF₄ + CLE; 2) TiF₄ + SBU; 3) CLE; 4) SBU. Cavities were restored using resin composite, and placed in intraoral palatal devices used by 16 volunteers for 21 days, to induce caries formation in situ. The fragments were then ground-flat to perform Knoop microhardness tests. Nine indentations were performed on each enamel and dentin substrate, subjacent to the restoration. Analysis of variance and Tukey’s test were applied.ResultsEnamel: groups receiving TiF₄ dentin pretreatment (regardless of adhesive system and tooth-restoration interface distance) presented higher hardness means at a depth of 25 μm from the outer tooth surface (p < 0.0001). Dentin: groups receiving CLE presented higher means when applying TiF₄ pretreatment, whereas groups restored with SBU presented higher means without pretreatment (p = 0.0003).ConclusionsDentin pretreatment with TiF₄ inhibited demineralization of the enamel interface in situ, regardless of the adhesive, and TiF₄ pretreatment followed by CLE application showed higher potential for inhibiting dentin demineralization at the interface.     

Inhibition of enamel mineral loss by fissure sealant: An in situ study

ObjectivesThis study evaluated the effect of fluoride and non-fluoride sealants on hardness decrease (HD) and marginal adaptation (MA) on enamel substrates after cariogenic challenge.MethodsOcclusal enamel blocks, from human third molars, were randomly divided into six groups (n = 12), according to occlusal fissures condition (S – sound; C – caries-like lesion; CF – caries-like lesion + topical fluoride) and sealants (F – FluroShield; H – Helioseal Clear Chroma). Lesion depths were 79.3 ± 33.9 and 61.3 ± 23.9 for C and CF groups, respectively. Sealants were placed on occlusal surface and stored at 100% humidity (37 °C; 24 h/d). HD was measured by cross-sectional microhardness analysis at the sealant margin distances: ?1 (under sealant), 0 (sealant margin), 1, 2 (outer sealant). Sealant MA was observed by polarized light microscopy and scored according to: 0 – failure (no sealant MA or total sealant loss); 1 – success (sealant MA present). MA and HD were analysed by ANOVA-R and mixed model analysis, respectively.ResultsFor HD (ΔS), F values (6900.5 ± 3686.6) were significantly lower than H values (8534.6 ± 5375.3) regardless of enamel substrates and sealant margin distances. Significant differences were observed among sealant margin distances: ?1 (5934.0 ± 3282.6) < 0 (8701.5 ± 6175.7) = 1 (8473.2 ± 4299.4) = 2 (7761.5 ± 4035.1), regardless of sealant and substrate. MA was similar for all groups (p ≥ 0.05).ConclusionMA was not affected by sealant type or substrate condition, whereas enamel HD was favourably impacted by fluoride in the sealant. In addition, sealants were more effective as a physical barrier than as its chemical potency in reducing enamel HD.Clinical significanceSealing with a fluoride material is a recommended procedure to prevent caries of occlusal permanent molars in high-caries-risk patients, even though those exhibiting white spot lesions, since the enamel hardness decrease when fluoride sealant was used in vitro.     

Effect of biomimetic mineralization on enamel and dentin: A Raman and EDX analysis

ObjectiveTo investigate the effect of an experimental biomimetic mineralization kit (BIMIN) on the chemical composition and crystallinity of caries-free enamel and dentin samples in vitro.MethodsEnamel and dentin samples from 20 human teeth (10 for enamel; 10 for dentin) were divided into a control group without treatment and test samples with BIMIN treatment. Quantitative analysis of tissue penetration of fluoride, phosphate, and calcium was performed using energy-dispersive X-ray spectroscopy (EDX). Mineralization depth was measured by Raman spectroscopy probing the symmetric valence vibration near 960 cm⁻¹ as a marker for crystallinity. EDX data was statistically analyzed using a paired t-test and Raman data was analyzed using the Student’s t-test.ResultsEDX analysis demonstrated a penetration depth of fluoride of 4.10 ± 3.32 μm in enamel and 4.31 ± 2.67 μm in dentin. Calcium infiltrated into enamel 2.65 ± 0.64 μm and into dentin 5.58 ± 1.63 μm, while the penetration depths for phosphate were 4.83 ± 2.81 μm for enamel and 6.75 ± 3.25 μm for dentin. Further, up to 25 μm of a newly mineralized enamel-like layer was observed on the surface of the samples. Raman concentration curves demonstrated an increased degree of mineralization up to 5–10 μm into the dentin and enamel samples.SignificanceBiomimetic mineralization of enamel and dentin samples resulted in an increase of mineralization and a penetration of fluoride into enamel and dentin.     

Effect of fluoride toothpaste with nano-sized trimetaphosphate on enamel demineralization: An in vitro study

ObjectiveThis study evaluated the effect of toothpastes containing 1100 ppm F associated or not with micrometric or nano-sized sodium trimetaphosphate (TMP) on enamel demineralization in vitro, using a pH cycling model.DesignBovine enamel blocks (4 mm × 4 mm, n = 96) were randomly allocated into eight groups (n = 12), according to the test toothpastes: Placebo (without fluoride or TMP); 1100 ppm F (1100F); 1100F plus micrometric TMP at concentrations of 1%, 3% or 6%; and 1100F plus nanosized TMP at 1%, 3% or 6%. Blocks were treated 2×/day with slurries of toothpastes and submitted to a pH cycling regimen for five days. Next, final surface hardness (SHf), integrated hardness loss (IHL), differential profile of integrated hardness loss (ΔIHL) and enamel fluoride (F) concentrations were determined. Data were analyzed by ANOVA and Student-Newman-Keuls’ test (p < 0.05).ResultsThe use of 1100F/3%TMPnano led to SHf 30% higher (p < 0.001) and IHL ∼ 80% lower (p < 0.001) when compared to 1100F. This toothpaste also resulted in ∼64% reduction of mineral loss (ΔIHL) when compared to 1100F. Moreover, the addition of nano-sized TMP promoted increases in enamel F uptake of 90%, 160% and 100%, respectively for the concentrations of 1%, 3% and 6%, when compared to 1100F (p < 0.001).ConclusionThe addition of nano-sized TMP at 3% to a conventional toothpaste significantly decreased enamel demineralization when compared to its counterparts without TMP or supplemented with micrometric TMP.     

Examination of ceramic/enamel interfacial debonding using acoustic emission and optical coherence tomography

ObjectiveThis study investigates monitored micro-crack growth and damage in the ceramic/enamel adhesive interface using the acoustic emission (AE) technique with optical coherence tomography (OCT) under fatigue shear testing.MethodsShear bond strength (SBS) was measured first with eight prepared ceramic/enamel adhesive specimens under static loads. The fatigue shear testing was performed with three specimens at each cyclic load according to a modified ISO14801 method, applying at 80%, 75%, 70%, and 65% of the SBS to monitor interface debonding. The number of cycles at each load was recorded until ceramic/enamel adhesive interface debonding occurred. The AE technique was used to detect micro-crack signals in static and fatigue shear bond tests.ResultsThe results showed that the average SBS value in the static tests was 18.07 ± 1.72 MPa (mean ± standard deviation), expressed in Newton's at 56.77 ± 5.40 N. The average number of fatigue cycles in which ceramic/enamel interface damage was detected in 80%, 75%, 70% and 65% of the SBS were 41, 410, 8141 and 76,541, respectively. The acoustic behavior varied according to the applied load level. Events were emitted during 65% and 70% fatigue tests. A good correlation was observed between the crack location in OCT images and the number of AE signal hits.SignificanceThe AE technique combined with OCT images as a pre-clinical assessment tool to determine the integrity of cemented load bearing restored ceramic material. Sustainable cyclic load stresses in ceramic/enamel bonded specimens were substantially lower than the measured SBS. Predicted S–N curve showed that the maximum endured load was 10.98 MPa (about 34.48 N) passing 10⁶ fatigue cyclic.     

A dual energy micro-CT methodology for visualization and quantification of biofilm formation and dentin demineralization

ObjectiveThe aim of this study was to induce artificial caries in human sound dentin by means of a microcosm model using human saliva as source of bacteria and to apply a novel dual-energy micro-CT technique to quantify biofilm formation and evaluate its demineralization potential.DesignEight sound third molars had the occlusal enamel removed by cutting with a diamond disk and five cylindrical cavities (±2 mm diameter; ±1.5 mm depth) were prepared over the dentin surface in each specimen (n = 40 cavities). After sterilization, each specimen received the bacterial salivary inoculum obtained from individuals without any systemic diseases presenting dentin caries lesions and were incubated in BHI added of with 5% sucrose for 96 h to allow biofilm formation. After that, two consecutive micro-CT scans were acquired from each specimen (40kv and 70kv). Reconstruction of the images was performed using standardized parameters. After alignment, registration, filtering and image calculations, a final stack of images containing the biofilm volume was obtained from each prepared cavity. Dentin demineralization degree was quantified by comparison with sound dentin areas. All data were analyzed using Shapiro-Wilk test and Spearman correlation using α=5%.ResultsDual-energy micro-CT technique disclosed biofilm formation in all cavities. Biofilm volume inside each cavity varied from 0.30 to 1.57 mm³. A positive correlation between cavity volume and volume of formed biofilm was obtained (0.77, p < 0.01). The mineral decrease obtained in dentin was high (± 90%) for all cavities and all demineralized areas showed mineral density values lower than a defined threshold for dentin caries (1.2 g/cm³).ConclusionDual-energy micro-CT technique was successful in the quantification of a microcosm human bacterial biofilm formation and to quantify its demineralization potential in vitro.     

In vitro enamel erosion and abrasion-inhibiting effect of different fluoride varnishes

ObjectiveTo investigate the erosion and abrasion inhibiting effect of CPP-ACP/NaF and xylitol/NaF varnishes.MethodsBovine enamel samples (n = 40) were exposed to the following treatments (n = 10): NaF varnish (Duraphat^®, positive control); CPP-ACP/NaF varnish (MI varnishTM); xylitol/NaF (Profluorid^®) or distilled and deionized water (MilliQ^®, negative control). The samples were submitted for 3 days to 4 cycles/day of erosion (5 min in Sprite Zero) and 2 cycles of abrasion/day after the first and last erosive challenge, with a toothbrush machine and slurries of a placebo toothpaste for 15 s (50 strokes/s). Among the cycles and after the last daily cycle, the specimens remained in artificial saliva. The change in the enamel surface was evaluated by using 3D non-contact optical profilometry with surface roughness (Ra and Sa values) and tooth structure loss (TSL) measurements. Scanning electron microscopy (SEM) assessed the enamel topographic characteristics. Differences in the Ra, Sa and TSL among treatments were tested using one-way ANOVA followed by the Tukey test.ResultsAll varnishes promoted better results for Ra and Sa values than the negative control (p = 0.0001), without difference among them (p > 0.05). However, CPP-ACP/NaF varnish stimulated fewer TSL (7.09 ± 0.70 μm) compared to NaF varnish (10.33 ± 1.36 μm, p = 0.002), xylitol/NaF varnish (9.96 ± 0.41 μm, p = 0.007) and the negative control (18.38 ± 3.32 μm, p = 0.0001).ConclusionA single-application of fluoride topical varnishes was effective in reducing enamel wear. The CPP-ACP/NaF varnish had the best effect against enamel loss from an erosion-abrasion challenge.     

Amazonian delicacy tucupi is as erosive as a cola-based soft drink

ObjectiveAcidic diets are advocated as main risk factor for tooth erosion, which could be prevented, or at least controlled, if patients were early advised. It is important to identify, hence, if possible dietary constituents regionally consumed on large scale, such as tucupi, a low-pH yellowish-green color and strong flavor delicacy made from the juice of a bitter cassava, may explain its occurrence in specific patient groups. This cross-over in situ/ex vivo study evaluated tucupi's ability to promote erosion of bovine enamel by assessing its percentage of surface microhardness change (%SMHC), taking a cola-based soft drink and human saliva as positive and negative controls.DesignFor three 7-days spaced out legs of 7 days each, nine volunteers wore palatal devices with three bovine enamel blocks, which were challenged with one of the following solutions: TUC—tucupi (n = 27); COL—cola-based soft drink (n = 27); SAL—saliva (n = 27). Erosive challenges were performed extra-orally (4×/day) by dropping TUC or COL at room temperature on specimens. After 5 min, palatal devices were replaced into the mouth. SAL permanently acted as the negative control while volunteers solely wore the device. One-way ANOVA followed by Tukey’s post-hoc tests (α = 0.05) were applied.ResultsTUC promoted an enamel %SMHC (−21.56 ± 10.08^a) similar than that promoted by COL (−18.19 ± 12.99^a; p = 0.275), which were both significantly higher than that promoted by SAL (−1.86 ± 13.65^b; p < 0.0001).ConclusionsBesides the most worldwide appreciated cola-based soft drink, the greatly consumed Amazonian delicacy tucupi can be considered a potential risk factor for tooth erosion.     

Sodium fluoride effect on erosion–abrasion under hyposalivatory simulating conditions

ObjectivesTo investigate the effect of fluoride (0, 275 and 1250 ppm F; NaF) in combination with normal and low salivary flow rates on enamel surface loss and fluoride uptake using an erosion–remineralization–abrasion cycling model.DesignEnamel specimens were randomly assigned to 6 experimental groups (n = 8). Specimens were individually placed in custom made devices, creating a sealed chamber on the enamel surface, connected to a peristaltic pump. Citric acid was injected into the chamber for 2 min followed by artificial saliva at 0.5 (normal flow) or 0.05 (low flow) ml/min, for 60 min. This cycle was repeated 4×/day, for 5 days. Toothbrushing with abrasive suspensions containing fluoride was performed for 2 min (15 s of actual brushing) 2×/day. Surface loss was measured by optical profilometry. KOH-soluble fluoride and enamel fluoride uptake were determined after the cycling phase. Data were analysed by two-way ANOVA.ResultsNo significant interactions between fluoride concentration and salivary flow were observed for any tested variable. Low caused more surface loss than normal flow rate (p < 0.01). At both flow rates, surface loss for 0 was higher than for 275, which did not differ from 1250 ppm F. KOH-soluble and structurally-bound enamel fluoride uptake were significantly different between fluoride concentrations with 1250 > 275 > 0 ppm F (p < 0.01).ConclusionsSodium fluoride reduced enamel erosion/abrasion, although no additional protection was provided by the higher concentration. Higher erosion progression was observed in low salivary flow rates. Fluoride was not able to compensate for the differences in surface loss between flow rates.     

Influence of bonding surface and bonding methods on the fracture resistance and survival rate of full-coverage occlusal veneers made from lithium disilicate ceramic after cyclic loading

ObjectivesThe purpose of this laboratory study was to evaluate the influence of bonding method and type of dental bonding surface on fracture resistance and survival rate of resin bonded occlusal veneers made from lithium disilicate ceramic after cyclic loading.MethodsFourty-eight extracted molars were divided into three groups (N = 16) depending on the preparation: within enamel, within dentin/enamel or within enamel/composite resin filling. Lithium disilicate occlussal veneers were fabricated with a fissure-cusp thickness of 0.3–0.6 mm. Restorations were etched (5% HF), silanated and adhesively luted using a dual-curing luting composite resin. Test groups were divided into two subgroups, one using a only a self-etching primer, the other additionally etching the enamel with phosphoric acid. After water storage (37 °C; 21 d) and thermocycling (7500 cycles; 5–55 °C), specimens were subjected to dynamic loading in a chewing simulator (600,000 cycles; 10 kg/2 Hz). Surviving specimens were loaded until fracture using a universal testing machine.ResultsAll specimens survived artificial aging, several specimens showed some damage. ANOVA revealed that enamel etching provided statistically significantly (p ≤ 0.05) higher fracture resistance than self-etching when bonding to enamel and dentin. Self-etching provided statistically significant (p ≤ 0.05) higher fracture resistance for the enamel-composite group than for the enamel group. Enamel etching provided statistically significant (p ≤ 0.05) higher fracture resistance for the enamel and dentin group than for groups enamel and enamel-composite.SignificanceEtching enamel improved the fracture resistance of occlusal veneers when bonding to dentin and enamel and increased the survival rate when bonding to enamel.     

In vitro longitudinal evaluation of enamel wear by cross-polarization optical coherence tomography

ObjectivesEnamel thickness determination by Cross-Polarization Optical Coherence Tomography (CP-OCT) is a promising approach for quantitative monitoring of tooth wear progression. This study evaluated the ability of CP-OCT to quantify the thickness of natural enamel before, during and after tooth wear simulation.Materials and MethodsNatural, unpolished human dental enamel slabs were submitted to five wear stages (Wear 1: to level the surfaces; Wear 2 to Wear 5: 0.05 ± 0.02 mm reduction each) simulated by an automatic grinding/polishing machine. Enamel thickness was evaluated with CP-OCT and a gold-standard method (micro-CT) at baseline and after every wear stage. Data were analyzed using ANOVA with pairwise comparisons for wear stages’ impact on the thickness and wear depth measurements. The inter-method agreement was analyzed using intra-class correlation coefficients, the difference between means, and Bland-Altman plots.ResultsEnamel thickness measurements (mean ± standard error, in mm) with natural (1.40 ± 0.05) and worn surfaces (1.08 ± 0.02) by CP-OCT did not differ significantly from those measured by micro-CT (natural = 1.39 ± 0.05; worn = 1.09 ± 0.02; p-values = 0.30 and 0.39, respectively). CP-OCT and micro-CT showed excellent agreement on natural (ICC = 0.98) and worn surfaces (ICC = 0.98) enamel thickness measurements. Among and between wear stages, there were significant differences in enamel thickness and wear depth measurements for both methods (p-value <0.0001 for all). Both methods yielded similar measurements’ mean (0.14 ± 0.01; p-value = 0.87) and were in good agreement (ICC = 0.77) for wear depth estimation.SignificanceCP-OCT allows accurate measurement of enamel thickness on natural tooth surfaces. Enamel thickness measurement by CP-OCT allows quantitative monitoring of enamel thickness changes and wear depth following progressive wear.     

Frequency of intake and amount of fluoride in milk for remineralisation of artificial caries on enamel and dentine: Ex vivo/in situ study

ObjectivesThis study analysed the effect of frequency of intake and amount of fluoride in milk on the remineralisation of artificial enamel and dentine caries lesions ex vivo/in situ.Materials and methodsPre-demineralised bovine enamel and dentine slabs were randomly allocated into 5 groups and fixed in removable appliances used by subjects for 7 days in each phase. Each treatment comprised milk containing 2.5 ppm fluoride daily (T1), or every other day (T2), 5.0 ppm F daily (T3), or every other day (T4) or no treatment (T5).ResultsEnamel alterations were quantified by surface hardness recovery (%SHR) and transversal microradiography (TMR), and in dentine by TMR only. Data were analysed by ANOVA and Tukey’s test (p < 0.05). For enamel, the highest %SHR was found for T1 and T3 compared to control, without significant differences between them. All groups showed positive values of ΔΔZ − T1 (247.3 ± 198.5); T2 (110.9 ± 303.2); T3 (226.0 ± 299.2); T5 (5.0 ± 288.0), except T4 (−274.5 ± 407.3). For dentine, the only group that presented remineralisation was T2 (350.0 ± 657.5).ConclusionsFluoridated milk daily seems to have higher remineralising effect on enamel than its use every other day. Dentine, does not seem to benefit from daily use of fluoridated milk.     

Effect of eugenol-containing and resin endodontic sealers on retention of prefabricated metal posts cemented with zinc phosphate and resin cements

PurposeThe aim of this study was to compare the effects of two endodontic sealers on the retention of posts cemented with zinc phosphate or resin cement.Materials and methodsCrowns of 72 mandibular premolars were removed at the cementoenamel junction. Root canals were prepared and specimens were randomly divided into two groups of 36. In each group, 12 specimens were obturated with gutta percha only; 12 specimens with gutta percha/ZOE sealer and 12 specimens with gutta percha/AH26. In the first group, 10 mm Post spaces were prepared with Peeso reamers size 4 and, size 5 stainless steel Paraposts were cemented in with zinc phosphate. In the second group, 10 mm Post spaces were prepared with Fiber Lux size 5.5 drills and size 5 Paraposts were cemented with Panavia F_2.0. After mounting in resin blocks, posts were pulled out by universal testing machine at 1 mm/min and results were analyzed by two-way ANOVA and Dunnett test.ResultsMean forces (in Newtons) required to remove posts cemented with zinc phosphate in canals obturated without sealer, with ZOE, and with AH26 sealers were 270 ± 83, 281 ± 128 and 266 ± 67, respectively; and for posts cemented with Panavia F_2.0 were 520 ± 290, 464 ± 212 and 229 ± 108, respectively. Statistical analysis showed that AH26 significantly reduced retention of posts cemented with Panavia F_2.0 (p < 0.05).ConclusionDifferent sealers had no significant effect on retention of posts cemented with zinc phosphate. However posts cemented with Panavia F_2.0 showed reduced retention in canals obturated with AH26.