The effect of active components from citrus fruits on dentin MMPs

ObjectivesThis study was aimed to evaluate the anti-matrix metalloproteinases (MMPs) ability of active components from citrus fruits (hesperetin: Hst, hesperidin: Hsd and naringenin: Nge).MethodsInactivation effects of citrus flavonoids (Hst, Hsd, Nge) at different concentrations on soluble collagenase were measured using a fluorometric assay. Matrix-bound endogenous MMPs activity was evaluated via dry mass loss and hydroxyproline (HYP) release of demineralized human dentin. Demineralized dentin beams were pretreated with 500 μg/mL citrus flavonoids for 10 min. Chlorhexidine (CHX) was used as inhibitor control. Beams pretreated with distilled water served as blank control. Dentin slabs were used for in situ zymography and evaluated under confocal microscopy. Ultrastructure of demineralized collagen fibers was exhibited by Transmission Electron Microscopy (TEM).ResultsCitrus flavonoids exhibited inactivation function on soluble MMPs and the extent of inactivation increased in a dose-dependent manner. The inactivation percent of citrus flavonoids reached above 90% at the concentration of 500 μg/mL. Compared with control group, citrus flavonoids pretreated demineralized dentin beams exhibited less dry mass loss, lower hydroxyproline release and more intact collagen architecture after 15 days storage. Dentin samples pretreated with citrus flavonoids showed lower enzymes activities in in situ zymography.ConclusionsHst, Hsd or Nge have anti-MMPs ability and can preserve dentin collagen from degradation.Clinical Significance: Hst, Hsd and Nge may have the potential to be used in dentin bonding systems and improve the resin-dentin bonding durability.     

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.     

Role of proteoglycans on the biochemical and biomechanical properties of dentin organic matrix

ObjectiveProteoglycans (PGs) are multifunctional biomacromolecules of the extracellular matrix of collagen-based tissues. In teeth, besides a pivotal regulatory role on dentin biomineralization, PGs provide mechanical support to the mineralized tissue and compressive strength to the biosystem. This study assessed enzymatic protocols for selective PGs removal from demineralized dentin to determine the roles of these biomacromolecules in the bulk mechanical properties and biostability of type I collagen.MethodsSelective removal of glycosaminoglycans chains (GAGs) and PGs from demineralized dentin was carried out by enzymatic digestion protocols using chondroitinase ABC (c-ABC) and trypsin (Try). A comprehensive study design included assessment of dentin matrix mass loss, biodegradability of the PGs/GAGs-depleted dentin matrix, ultimate tensile strength (UTS) and energy to fracture tests. Quantitative data was statistically analyzed by two-way and one-way ANOVA followed by the appropriate post hoc tests (α = 0.05).ResultsTransmission electron microscopy images show effective GAGs removal by c-ABC and Try and both enzymatic methods released statistically similar amounts of GAGs from the demineralized dentin. Try digestion resulted in about 25% dentin matrix mass loss and increased susceptibility to collagenolytic digestion when compared to c-ABC (p = 0.0224) and control (p = 0.0901). Moreover, PGs digestion by Try decreased the tensile strengths of dentin. Statistically lower energy to fracture was observed in c-ABC-treated dentin matrix.ConclusionsGAGs plays a pivotal role on tissue mechanics and anisotropy, while the core protein of PGs have a protective role on matrix biostability.     

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.     

Inhibition of endogenous human dentin MMPs by Gluma

ObjectiveThe 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.MethodsDentin 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.ResultsGluma 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.SignificanceThe use of Gluma may contribute to the preservation of adhesive interfaces by its cross-linking and inhibitory properties of endogenous dentin MMPs.     

Alternative model for cathepsin K activation in human dentin

ObjectiveTo evaluate the protease activity in dentin matrices subjected to lactic acid (LA) in comparison to polyacrylic acid (PAA) challenge model at cathepsin K (CT-K) optimum pH 5.5 to assess effectiveness of inhibitors in dentin collagen degradation.MethodsDentin disks measuring 0.5 mm prepared from human molars were completely demineralized in 10% H₃PO₄. Demineralized dentin disks were challenged with 0.1 M LA, 1.1 mM PAA, artificial saliva (AS), or deionized water (C) for 24 h or 7-days. Dentin collagen properties were tested by measurement of %dry mass change, and ultimate tensile strength (UTS). Degradation of dentin type I collagen was measured by telopeptide assays measuring the sub-product release of C-terminal cross-linked telopeptides (ICTP) and C-terminal peptide (CTX) in the incubation media in relation to total protein concentration, which correlates with matrix metalloproteinases (MMPs) and CT-K activities.ResultsGravimetric analysis showed statistically significant difference between C and other groups (p < 0.04) at 24 h. LA specimens showed significantly higher weight loss from 24 h to 7-days (p = 0.02). UTS revealed statistically significant difference between AS and LA at 24 h and 7-days. UTS at 24 h and 7-days for C and AS had significantly higher mean values compared to LA and PAA. Telopeptide assays reported that CTX_tp results showed that LA at 24 h had significantly higher mean values compared to C and AS.SignificanceLA has the ability to activate endogenous CT-K in dentin as measured by the release of CTX (CT-K specific telopeptide). This LA based model has the potential application for further investigations on the activity and possible inhibitors of CT-K in human dentin.     

Distinct effects of polyphenols and solvents on dentin collagen crosslinking interactions and biostability

ObjectiveTo evaluate the effects of different polyphenols and solvents on dentin collagen’s crosslinking interactions and biostabilization against MMPs and collagenase degradation.MethodsTwo polyphenols [proanthocyanidin (PA) and quercetin (QC)] with different water solubility were prepared as treatment solutions using ethanol (EtOH) or dimethyl sulfoxide (DMSO) as solvents. 6-um-thick dentin films were microtomed from dentin slabs of third molars. Following demineralization, films or slabs were subject to 60-s treatment (PA or QC) or no treatment (control) with subsequent extended-rinse with original solvent (EtOH or DMSO) or distilled water (DW). Collagen crosslinking interactions were assessed by FTIR. Biostability was assessed through endogenous MMPs activity via confocal laser scanning microscopy, and exogenous collagenase degradation via weight loss, hydroxyproline release and SEM. Finally, direct collagenase inactivation was also evaluated. Data were analyzed by three-way ANOVA and post-hoc tests (α = 0.05%).ResultsDistinct effects of two polyphenols and solvents on collagen crosslinking and biostabilization were observed. Higher crosslinking and biostability efficacy occurred with PA than QC (p < 0.001) that demonstrated negligible collagen interactions. With DMSO solvent, efficacy results were significantly reduced with both polyphenols (p < 0.05). DMSO-rinse further weakened interactions of PA with collagen, diminishing biostability (p < 0.05). Low biostability was detected with QC and DW-rinse, suggesting direct enzymatic inhibition due to physical presence in collagen.SignificanceCollagen crosslinking interactions and biostability depend on polyphenol chemical characteristics. Treatment-solution solvents may affect interactions between polyphenols and collagen, specifically, DMSO showed detrimental effects on collagen crosslinking and biostability and should be used with caution.     

Investigation into the validity of WearCompare,a purpose-built software to quantify erosive tooth wear progression

ObjectivesThe use of surface matching software with intraoral scanners is developing rapidly which increases the need for accessible, accurate and validated measurement software. This investigation compared the current gold-standard Geomagic Control software to a purpose-built software “WearCompare”.MethodsArtificially created occlusal defects of a known size were created on 10 natural molar teeth scanned with a structured-light model-scanner (Rexcan DS2, Europac 3D, Crewe). The volume change, maximum profilometric loss and mean profilometric loss were obtained from both Geomagic Control (3D Systems, Darmstadt, Germany) and WearCompare (leedsdigitaldentistry.com). Duplicated datasets were randomly repositioned and re-alignment performed. The effect of the re-alignment was calculated by analysing differences between the known defect size and defect size after re-alignment using the same measurement metrics. Lastly, clinical wear measurements were compared on natural molar surfaces (n = 60) over 6 months using study models collected from a previous longitudinal trial. Data analysis was performed in SPSS v25 (paired t-tests, Pearson correlations, p < 0.05).ResultsMeasurement correlation between the softwares was greater than 0.97 (p < 0.001) for all measurement metrics. The volume change error (SD) after alignment was −0.67 mm³(1.14) for Geomagic and −0.06 mm³(0.93) for WearCompare (p = 0.140 and r = 0.065, p = 0.86). Measurement errors were observed after alignment in both softwares and no statistical differences were observed between softwares. The volume change on the clinical dataset over 6 months was +0.29 mm³(3.97) in Geomagic and −0.30 mm³(1.82) for WearCompare (p = 0.19 and r = 0.61, p < 0.001). The mean profile gain was 42.86 μm(40.19) for Geomagic and 32.17 μm(23.72) for WearCompare (p = 0.048). Correlations between the softwares were greater than 0.6 for all measurement metrics except for mean profile gain.SignificanceWearCompare is a comparable tool to Geomagic for quantifying erosive tooth wear. WearCompare reported statistically less profile gain indicating less error but further research is needed to reduce the human errors in both softwares.     

Promotion of enamel caries remineralization by an amelogenin-derived peptide in a rat model

ObjectiveAn amelogenin-derived peptide has been shown to promote remineralization of demineralized enamel in an in vitro model of initial caries induced by pH cycling. The present study examines whether the peptide exerts similar effects within the complex oral environment in vivo.DesignSpecific pathogen-free Sprague-Dawley rats (n = 36) were infected with Streptococcus mutans, given ad libitum access to Diet 2000 and drinking water supplemented with sucrose (10%, w/v), and then randomly divided into three groups treated with 25 μM peptide solution, 1 g/L NaF or deionized water. Molar teeth were swabbed twice daily with the respective solutions for 24 days. Then animals were killed, their jaws were removed and caries lesions were analyzed using the quantitative light-induced fluorescence-digital (QLF-D) technique to measure changes in mineral content. To verify QLF-D results, caries were scored for lesion depth and size using the Keyes method, and analyzed using polarized light microscopy (PLM).ResultsMineral gain was significantly higher in teeth treated with peptide or NaF than in teeth treated with water (p < 0.05), based on the QLF-D results (ΔF and ΔQ). Incidence of smooth-surface and sulcal caries based on Keyes scores was similar in rats treated with peptide or NaF, and significantly lower in these groups than in rats treated with water (p < 0.05). Lesions on teeth treated with peptide or NaF were shallower, based on PLM. No significant differences were observed between molar enamel caries treated with peptide or NaF.ConclusionsThis amelogenin-derived peptide can promote remineralization in a rat caries model, indicating strong potential for clinical use.     

Stability of bonds made to superficial vs. deep dentin,before and after thermocycling

ObjectivesBonding stability of resinous adhesives to dentin is still problematic and may involve regional variations in dentin composition. This study is to evaluate the effect of dentin depth on the stability of resin-dentin bonds under thermocycling challenge.MethodsDentin slabs with two flat surfaces parallel to the tooth axis were obtained from extracted human third molars. The slabs were randomized into eight groups according to the location of dentin [deep dentin (DD) or superficial dentin (SD)], the adhesive treatment (Single Bond 2 or Clearfil S³ Bond), and the storage treatment (thermocycling for 5000 times vs. no). After the adhesive treatment and composite buildup on the dentin slabs, the micro-shear bond strength (μSBS) of each group was detected. The concentrations of cross-linked carboxyterminal telopeptide of type I collagen (ICTP) were also evaluated using an immunoassay to detect the degree of collagen degradation in each group.ResultsDentin depth, adhesive treatment and storage treatment all showed significant effects on both the μSBSs and the ICTP values (P < 0.05). Regardless of the adhesive type, thermocycling decreased the μSBSs and increased the ICTP values (P < 0.05). The DD groups showed significantly lower μSBSs and higher ICTP values than SD groups after thermocycling aging (P < 0.05). The treatment with Single Bond 2 significantly increased the ICTP values (P < 0.05), whereas Clearfil S³ Bond showed no effect on the ICTP values (P > 0.05).SignificanceDeep dentin showed significantly more bond degradation after thermocycling than did superficial dentin.     

Distinct decalcification process of dentin by different cariogenic organic acids: Kinetics,ultrastructure and mechanical properties

ObjectivesWe studied artificial dentin lesions in human teeth generated by lactate and acetate buffers (pH 5.0), the two most abundant acids in caries. The objective of this study was to determine differences in mechanical properties, mineral density profiles and ultrastructural variations of two different artificial lesions with the same approximate depth.Methods0.05 M (pH 5.0) acetate or lactate buffer was used to create 1) 180 μm-deep lesions in non-carious human dentin blocks (acetate 130 h; lactate 14days); (2) demineralized, ∼180 μm-thick non-carious dentin discs (3 weeks). We performed nanoindentation to determine mechanical properties across the hydrated lesions, and micro X-ray computed tomography (MicroXCT) to determine mineral profiles. Ultrastructure in lesions was analyzed by TEM/selected area electron diffraction (SAED). Demineralized dentin discs were analyzed by small angle X-ray scattering (SAXS).ResultsDiffusion-dominated demineralization was shown based on the linearity between lesion depths versus the square root of exposure time in either solution, with faster kinetics in acetate buffer. Nanoindentation revealed lactate induced a significantly sharper transition in reduced elastic modulus across the lesions. MicroXCT showed lactate demineralized lesions had swelling and more disorganized matrix structure, whereas acetate lesions had abrupt X-ray absorption near the margin. At the ultrastructural level, TEM showed lactate was more effective in removing minerals from the collagenous matrix, which was confirmed by SAXS analysis.ConclusionsThese findings indicated the different acids yielded lesions with different characteristics that could influence lesion formation resulting in their distinct predominance in different caries activities, and these differences may impact strategies for dentin caries remineralization.     

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.     

Disinfection of dentinal tubules with 2% Chlorhexidine gel,Calcium hydroxide and herbal intracanal medicaments against Enterococcus faecalis: An in-vitro study

AimThis in vitro study was conducted to evaluate the disinfection of dentinal tubules using 2% Chlorhexidine gel, Honey, Aloe vera gel, Curcuma longa, Propolis gel and Calcium hydroxide against Enterococcus faecalis.Materials and methodTwo hundred and ten human mandibular first premolars were infected with Enterococcus faecalis for 21 days. Samples were divided into 7 groups. Group I- Saline (negative control), Group II- 2% Chlorhexidine gel(CHX), Group III- honey, Group IV- Aloe vera gel, Group V- 20% Curcuma longa gel, Group VI- Propolis gel and Group VII -Calcium hydroxide (CH). At the end of 1, 3 and 5 days, the antimicrobial efficacy of medicaments against E.faecalis was assessed at the depths of 200 µm and 400 µm.Results2% Chlorhexidine gel was most effective followed by Propolis and Curcuma longa.Conclusion2% Chlorhexidine gel gave the best results. Among the herbal extracts Propolis and Curcuma longa hold a promising future but to implement their use as sole intracanal medicaments clinically, further in vivo and long term studies are warranted.     

Demineralized dentin 3D porosity and pore size distribution using mercury porosimetry

ObjectivesThe objectives of this study were to assess demineralized dentin porosity and quantify the different porous features distribution within the material using mercury intrusion porosimetry (MIP) technique. We compared hexamethyldisilazane (HMDS) drying and lyophilization (LYO) (freeze-drying) in sample preparation.MethodsFifty-six dentin discs were assigned into three groups. The control (CTR) group discs were superficially acid-etched (15 s 37% H₃PO₄) to remove the smear layer and then freeze-dried whereas LYO and HMDS groups samples were first totally demineralized using EDTA 0.5 M and then freeze-dried and HMDS-dried respectively. MIP was used to determine open porosity and pore size distribution of each pair of samples. Field emission scanning electron microscopy (FESEM) was used to illustrate the results.ResultsThe results showed two types of pores corresponding either to tubules and micro-branches or to inter-fibrillar spaces created by demineralization. Global porosity varied from 59% (HMDS-dried samples) to 70% (freeze-dried samples). Lyophilization drying technique seems to lead to less shrinkage than HMDS drying. FESEM revealed that collagen fibers of demineralized lyophilized samples are less melted together than in the HMDS-dried samples.SignificanceDemineralized dentin porosity is a key parameter in dentin bonding that will influence the hybrid layer quality. Its characterization could be helpful to improve the monomers infiltration.     

Effect of natural gel product on bovine dentin erosion in vitro

Objective

To evaluate the efficacy of Neem (Azadirachta indica) experimental gel for the prevention of erosive wear on bovine dentin, in vitro.

Material and Methods

One hundred dentin blocks were allocated into 5 experimental groups (20 samples each): C (control group, without gel); CG (control group, only base gel); F (fluoride gel, 1.23% NaF; pH 4.1, Dentsply; Brazil); N (Neem gel, 10% neem extract; pH 4.1, manipulation); NF (Neem+fluoride gel, 10% Neem extract and 1.23% NaF; pH 4.1, manipulation). The blocks were stored in artificial saliva for 24 hours. After this, they were submitted to six alternating re- and demineralization cycles. The blocks were analyzed for wear (profilometry). The results were submitted to statistical analysis by ANOVA and Tukey tests (P<0.05).

Results

The mean wear (±SD, µm) was shown as follows in groups: C (13.09±0.99), CG (10.60±1.99), F (10.90±1.44), N (12.68±1.13) and NF (10.84±1.65). All gels showed some preventive action when compared with control group. However, significant differences were found only between Neem+fluoride gel and fluoride gel.

Conclusion

A single application of a neem-containing fluoride gel reduced dentin erosion, thus it is a possible alternative in reducing dental wear. Further research should investigate the action mechanism and the synergism between them.     

Effect of laser activated bleaching on the chemical stability and morphology of intracoronal dentin

ObjectivesTo evaluate the effect of the bleaching with 35% hydrogen peroxide either activated or not by a 970 nm diode laser on the chemical stability and dentin surface morphology of intracoronary dentin.MethodsTwenty-seven slabs of intracoronary dentin specimens (3 × 3 mm) were distributed into three groups (n = 9), according to surface treatment: HP – 35% hydrogen peroxide (1 × 4’), DL – 970 nm diode laser (1 × 30”/0,8W/10 Hz), HP + DL – 35% HP activated with 970 nm diode laser (1 × 30”/0,8W/10 Hz leaving the gel in contact to the surface for 4′ after activation). Three Raman spectra from each fragment were obtained to calculate the mean intensity of peaks of inorganic component (a.u.), organic collagen content (a.u.), and the ratio of inorganic/organic content, before and after treatment. Analyses of the samples by confocal laser microscopy were performed to evaluate the surface roughness, percentage of tubules, perimeter and area percentage of tubules, before and after treatment. Data were analyzed by Kruskal-Wallis, Dunn’s, and Wilcoxon test (P < 0.05).ResultsData analysis showed that HP + DL did not change the inorganic content peaks 8.31 [29.78] or the inorganic/organic ratio 3.37 [14.67] (P > 0.05). Similarly, DL did not affect the chemical stability of the dentin surface (P > 0.05). However, HP significantly increased inorganic content peaks 10.87 [22.62], as well as the inorganic/organic ratio 6.25 [27.78] (P < 0.05). Regarding the morphological alterations, all surface treatments increase tubules exposure; HP treatment significantly increases perimeter and area percentage; and HP + DL increases surface roughness.ConclusionsBleaching HP combined with DL offers an improvement in terms of intracoronal dentin surface protection, yielding better maintenance of dentin chemical stability and morphology.     

Effect of generation 4.0 polyamidoamine dendrimer on the mineralization of demineralized dentinal tubules in vitro

ObjectiveDentine hypersensitivity is a type of clinical oral disease, which is highly prevalent worldwide. Although there are many materials to treat dentine hypersensitivity, their long-term therapeutic effects are not satisfactory. Therefore, the aim of this research was to observe and identify the biological mineralization of the generation 4.0 polyamidoamine dendrimer on the demineralized dentinal tubules at different time points.Design2 mm-thick slices were obtained from the cemento-enamel junction of 36 third molar teeth that simulated the condition of sensitivity with acid etching. Slices were treated with generation 4.0 polyamidoamine dendrimer and peptide bond condensing agent, while no treatment was applied on the slices of the control group. Following immersion in artificial saliva for 2, 4, 6, and 8 weeks respectively, the mineralization condition of dentine slices was observed using the scanning electron microscope (SEM). In addition, the differences in the samples of dental slices between the 2 groups were also detected using the microhardness test.ResultsSEM results showed that the average diameter and density of the dentinal tubules in the experimental group were significantly lower than those in the control group (P < 0.001). The microhardness test exhibited a similar result, which suggested that the microhardness of the experimental group was significantly higher than the control group (P < 0.001).ConclusionGeneration 4.0 polyamidoamine dendrimer promotes the biomineralization of demineralized dentinal tubules. Moreover, this result also suggests that the 4.0th generation polyamidoamine dendrimer has the potential value for dentine hypersensitivity treatment.     

TiF4 and NaF varnishes as anti-erosive agents on enamel and dentin erosion progression in vitro

Objective

This study assessed the effect of fluoride varnishes on the progression of tooth erosion in vitro. Material and Methods: Forty-eight enamel and 60 root dentin samples were previously demineralized (0.1% citric acid, pH 2.5, 30 min), leading to a baseline and erosive wear of 12.9 and 11.4 µm, respectively. The samples were randomly treated (6 h) with a 4% TiF₄ varnish (2.45%F-, pH 1.0), a 5.42% NaF varnish (2.45%F-, pH 5.0), a placebo varnish and no varnish (control). The samples were then subjected to erosive pH cycles (4x90 s/day in 0.1% citric acid, intercalated with artificial saliva) for 5 days. The increment of the erosive tooth wear was calculated. In the case of dentin, this final measurement was done with and without the demineralized organic matrix (DOM). Enamel and dentin data were analyzed using ANOVA/Tukey’s and Kruskal-Wallis/Dunn tests, respectively (p<0.05).

Results

The TiF₄ (mean±s.d: 1.5±1.1 µm) and NaF (2.1±1.7 µm) varnishes significantly reduced enamel wear progression compared to the placebo varnish (3.9±1.1 µm) and control (4.5±0.9 µm). The same differences were found for dentin in the presence and absence of the DOM, respectively: TiF₄ (average: 0.97/1.87 µm), NaF (1.03/2.13 µm), placebo varnish (3.53/4.47 µm) and control (3.53/4.36 µm).

Conclusion

The TiF₄ and NaF varnishes were equally effective in reducing the progression of tooth erosion in vitro.     

Hydrogen peroxide induces cell proliferation and apoptosis in pulp of rats after dental bleaching in vivo: Effects of the dental bleaching in pulp

ObjectiveThis study provides an in vivo evaluation of the inflammatory response, levels of cell proliferation and apoptosis, and the presence of necrosis after dental bleaching with two concentrations of hydrogen peroxide (H₂O₂).DesignWistar rats were divided into Control (placebo gel), BLUE (20% H₂O₂, 1 × 50 min), and MAXX (35% H₂O₂, 3 × 15 min) groups. At 2 and 30 days, the rats were killed (n = 10). The jaws were processed for histology analysis and PCNA and Caspase-3-cleaved immunohistochemistry, and data were submitted to the Mann-Whitney or ANOVA test (P < 0.05).ResultsAt 2 days, the MAXX group showed necrosis and the BLUE group revealed moderate inflammation on the occlusal third of the crown (P < 0.05). At 30 days, tertiary dentin had formed and there was an absence of inflammation. The level of cell proliferation was higher in the middle third of the BLUE group (P < 0.05), and cervical of MAXX at 2 days (P < 0.05), decreasing at 30 days. The apoptosis was present at 2 days, particularly in the cervical third of the crown in the bleached groups (P < 0.05), with a decrease only at 30 days in the BLUE group (P < 0.05).ConclusionsThe concentration of H₂O₂ influences effects on the pulp tissue, where a higher concentration of H₂O₂ can cause necrosis in the pulp and a prolonged effect within the apoptotic process; lower concentrations of H₂O₂ provide moderate inflammation, cell proliferation and apoptosis with a reduction of these processes over time.     

The effect of magnesium hydroxide-containing dentifrice using an extrinsic and intrinsic erosion cycling model

ObjectiveTo evaluate, in vitro, the effect of Mg(OH)₂ dentifrice, and the influence of the number of experimental days, on the extrinsic (citric acid –CA) and intrinsic (hydrochloric acid –HCl) enamel erosion models.DesignHuman enamel slabs were selected according to surface hardness and randomly assigned to 3 groups (n = 9) as follows: non-fluoridated (negative control), NaF (1450 ppm F- positive control) and Mg(OH)₂ (2%) dentifrices. The slabs were daily submitted to a 2-h period of pellicle formation and, over a period of 5 days, submitted to cycles (3×/day) of erosive challenge (CA 0.05 M, pH = 3.75 or HCl 0.01 M, pH = 2 for 30 s), treatment (1 min −1:3 w/w of dentifrice/distilled water) and remineralization (artificial saliva/120 min). Enamel changes were determined by surface hardness loss (SHL) for each day and mechanical profilometry analysis. Data were analyzed by two-way ANOVA followed by Tukey’s test to % SHL and one-way ANOVA to profilometry (p < 0.05).ResultsThe number of experimental days influenced the erosion process for the two types of erosion models (p < 0.001). Mg(OH)₂-containing dentifrices were effective in reducing enamel extrinsic acid erosion as determined by % SHL (p < 0.001) when compared to the control group, being better than positive control (p < 0.001); however, the dentifrices were not effective for the intrinsic model (p = 0.295). With regards to surface wear, no statistically significant differences were found among the groups for CA (p = 0.225) and HCl (p = 0.526).ConclusionThe findings suggest that Mg(OH)₂ dentifrices might protect enamel against slight erosion, but protection was not effective for stronger acid erosion.