Inhibitory effect of Salvadora persica extract (Miswak) on collagen degradation in demineralized dentin: In vitro study.

BACKGROUND/PURPOSE: Root dentin is vulnerable to acid attack, suggesting a higher risk of demineralization than coronal enamel. This study aimed to evaluate the inhibitory effect of Miswak extract on collagen degradation of demineralized dentin lesion. MATERIALS AND METHODS: Demineralized bovine root dentin specimens were treated for 1 h by 20% Miswak extract and 0.12% Chlorehexidine (CHX) as a positive control group, and then subjected to collagenolytic attack (clostridium histolyticum 0.5 CDU/mL, 16 h). These cyclic treatments were repeated for 3 days. After the cyclic treatment, the images of the specimens were captured with a light microscope and the lesion depth of degraded collagen layer of all specimens was measured. The mean lesion depth was calculated and compared between the groups using descriptive and One-way ANOVA followed by Post hoc Tukey's tests. Significant level was set at p < 0.05. RESULTS: The mean lesion depth of CHX (28.6 ±â€¯3.37 µm) had the least value, followed by Miswak (37.5 ±â€¯4.01 µm) then the control (78.4 ±â€¯18.43 µm) group. There was a significant difference in the mean lesion depth among the three groups (p = 0.000). CONCLUSION: Miswak aqueous extract from S. persica was found to preserve the dentin collagen matrix from collagenase enzyme. This could be due to the organic compounds like flavonoids, saponins, alkaloids, tannins, and others which have been reported in literature. Present finding suggests that Miswak might play a positive effect in dentin caries prevention.

Effects of coating materials on nanoindentation hardness of enamel and adjacent areas.

OBJECTIVES: Materials that can be applied as thin coatings and actively release fluoride or other bioavailable ions for reinforcing dental hard tissue deserve further investigation. In this study we assessed the potential of resin coating materials in protection of underlying and adjacent enamel against demineralization challenge using nanoindentation. METHODS: Enamel was coated using Giomer (PRG Barrier Coat, PBC), resin-modified glass-ionomer (Clinpro XT Varnish, CXT), two-step self-etch adhesive (Clearfil SE Protect, SEP) or no coating (control). After 5000 thermal cycles and one-week demineralization challenge, Martens hardness of enamel beneath the coating, uncoated area and intermediate areas was measured using a Berkovich tip under 2mN load up to 200µm depth. Integrated hardness and 10-µm surface zone hardness were compared among groups. RESULTS: Nanoindentation and scanning electron microscopy suggested that all materials effectively prevented demineralization in coated area. Uncoated areas presented different hardness trends; PBC showed a remarkable peak at the surface zone before reaching as low as the control, while CXT showed relatively high hardness values at all depths. SIGNIFICANCE: Ion-release from coating materials affects different layers of enamel. Coatings with fluoride-releasing glass fillers contributed to reinforcement of adjacent enamel. Surface prereacted glass filler-containing PBC superficially protected neighboring enamel against demineralization, while resin-modified glass-ionomer with calcium (CXT) improved in-depth protection. Cross-sectional hardness mapping of enamel on a wide range of locations revealed minute differences in its structure.

Study on the influence of leucine-rich amelogenin peptide (LRAP) on the remineralization of enamel defects via micro-focus x-ray computed tomography and nanoindentation.

Regeneration of severely damaged enamel (e.g. deep demineralized lesions) is currently not possible, because the structural units of enamel crystal construction are removed after its maturation. The aim of this in vitro study was to evaluate the effect of surface impregnation by leucine-rich amelogenin peptide (LRAP) on the remineralization of eroded enamel using micro-focus x-ray computed tomography (µCT). Fifteen bovine enamel blocks were embedded in resin and three zones (sound, demineralization, and remineralization) were defined on each specimen. Lesions were prepared by immersing the samples in demineralization solution for 7 d. The samples were soaked in distilled water or 60 or 120 µg mL(-1) solution of LRAP in water for 30 min. After the surface treatment, specimens were incubated in artificial saliva for either 5 or 10 d at 37 °C. The amount of mineral gain (dΔZ%) and the relative changes in the lesion depth (dLD%), obtained from µCT, were used to evaluate the effect of LRAP on the remineralization of lesions. The effects of LRAP on cross-sectional integrated hardness ΔINH were studied after 10 d using nanoindentation. ANOVA test was used to determine the effect of time and/or LRAP concentration on dΔZ%, dLD% and ΔINH mean values. Tukey's analysis was used for multiple comparison testing (α = 0.05). Analysis of µCT data showed significant effect of time and LRAP concentration on the dΔZ% (p = 0.013, p = 0.003) and the dLD% (p < 0.001, p = 0.002) mean values. The nanoindentation hardness was significantly improved by 120 µg mL(-1) LRAP (p = 0.02). Also, the peptide treatment affected the mineral distribution throughout the lesion by inhibiting of superficial deposition. This study showed that the treatment of eroded lesions in enamel by LRAP can improve and regulate the pattern of remineralization in vitro.

Optical coherence tomography for evaluation of enamel and protective coatings.

Optical coherence tomography (OCT) is an interferometric imaging technique. This study aimed to employ OCT to evaluate four different resin-based materials including a coating containing glass-ionomer filler and calcium, a giomer, and two fluoride-releasing self-etch resins. The coating and its underlying and adjacent enamel were monitored using swept-source OCT (center wavelength: 1330 nm) at baseline, after 5,000 thermal cycles, and after 1, 4 and 7 days of demineralization (pH 4.5). The coatings showed different thicknesses (60-250 micrometers) and various levels of structural and interfacial integrity. OCT could detect a demineralization inhibition zone adjacent to the edge of the fluoride- and calcium-releasing material. Localized demineralization was occasionally observed under thinner coatings. Protection of susceptible enamel surfaces by thin resin-based bioactive coatings provides protection from demineralization. OCT can be used to non-destructively monitor the integrity of such coatings, as well as enamel changes beneath and adjacent to them.

Formation and characterization of hypermineralized zone beneath dentine lesion body induced by topical fluoride in-vitro.

OBJECTIVE: This in-vitro study aimed to evaluate and characterize the hypermineralized zone (Hyper-zone) formed beneath the remineralized dentine lesion body by transverse microradiography (TMR), scanning electron microscopy (SEM) and energy dispersive X-ray analysis (EDS). DESIGN: Demineralized bovine dentine specimens were treated with fluoride solutions (APF, NaF) and remineralized for 2-4 weeks. Then thin sections were prepared to characterize the Hyper-zone by TMR, EDS. Fractured specimen surfaces were observed by SEM. RESULTS: TMR analysis revealed a higher mineral density at Hyper-zone than that of sound dentine (48vol%) ranging from 50 up to 61vol% and the thickness ranging from 197 to 344µm for 4-week specimens, while specimens without fluoride treatment did not show Hyper-zone. SEM pictures at Hyper-zone showed no evident crystal-like deposits in dentinal tubules and no notable difference when compared to that in sound dentine. EDS analysis demonstrated higher concentrations of Ca and P at Hyper-zone than those in sound dentine, which corresponded to the TMR profile, while the magnesium (Mg) concentration was low at this zone. CONCLUSIONS: Demineralized dentine lesions exposed to fluoride and remineralization treatments exhibited Hyper-zone beneath the lesion body, in which the mineral density was higher than that of sound dentine. Possible mechanism for the formation of Hyper-zone was discussed by assuming removal of mineral regulators such as Mg and other organic substances from sound dentine during de-/remineralization processes.

Effects of curing mode and moisture on nanoindentation mechanical properties and bonding of a self-adhesive resin cement to pulp chamber floor.

OBJECTIVE: This study aimed to investigate the effect of dentin surface moisture and curing mode on microtensile bond strength (MTBS) and nanoindentation characteristics of a self-adhesive resin cement. METHODS: Forty-four extracted human molars were distributed into four groups according to dentin surface moisture (dry or wet) and curing mode of the resin cement (light or chemical). Clearfil SA Cement (Kuraray Noritake Dental, Japan) was used for cementation of composite cores to the pulp chamber dentin. The specimens were sectioned into beams for MTBS test at the pulpal floor. Nanoindentation hardness and creep of the cement layer were measured under 100 mN load with 30s hold segment. Data were statistically analyzed using two-way ANOVA and Weibull distribution of MTBS (α=0.05). RESULTS: Moisture, curing mode or their interaction did not significantly affect mean MTBS values that ranged 17.6-22.6 MPa (p>0.05); however, the lowest characteristic strength was found in moist chemically cured group. Hardness ranged 437-512 MPa, and was not affected by the experimental factors (p>0.05). Nanoindentation creep ranged 9.3-10.9% with the chemically cured groups showing the highest values, indicating lower cross-linking and deformation resistance of their polymer network. SIGNIFICANCE: Additional moisture on dentin surface did not contribute to adhesion of the anhydrous self-adhesive resin cement to dentin. Light-curing, despite attenuation through the composite core, was beneficial and improved nanoindentation creep resistance of the cement. The difference was not, however, reflected in the mean bond strength or hardness values.

Concurrent evaluation of composite internal adaptation and bond strength in a class-I cavity.

OBJECTIVES: This study investigated class-I cavity floor adaptation by swept-source optical coherence tomography (OCT) in combination with microtensile bond strength (MTBS) using different filling methods. METHODS: Two adhesive systems; Tokuyama Bond Force and Tri-S Bond Plus were used in conjunction with a universal composite (Estelite Sigma Quick) placed either incrementally (oblique) or in bulk with or without a flowable composite lining (Palfique Estelite LV). Ten serial B-scan images were obtained throughout each cavity by OCT (center wavelength: 1319nm). In order to evaluate adaptation defined as the cavity floor percentage showing no gap, a significant increase in the signal intensity was considered as gap at the bonded interface of the cavity floor. The same specimens were then cut into beams to measure MTBS at the cavity floor. RESULTS: Two-way ANOVA demonstrated that the interaction of adhesive systems and filling techniques was significantly affecting both adaptation and MTBS (p<0.05). There was a significant correlation between MTBS and adaptation at cavity floor (p<0.05). Cavity floor adaptation and MTBS were improved when incremental filling technique was applied, while the outcome of lining technique was variable. CONCLUSIONS: Quantitative assessment by OCT can non-destructively provide information on the performance and effectiveness of dental composites and restoration techniques. There was a moderate correlation between floor adaptation and bond strength in class-I cavities. Incremental application of composite restoration showed the best performance in terms of bond strength and internal adaptation. CLINICAL SIGNIFICANCE: Incremental application of composite restoration was the most advantageous placement technique in terms of bond strength and internal adaptation. The lack of placement pressure with flowable composites may affect their adaptation to all-in-one adhesives; therefore, the outcome of cavity lining by flowable composite was variable.

Effects of structural orientation of enamel and dentine on light attenuation and local refractive index: an optical coherence tomography study.

OBJECTIVES: To investigate optical properties of human enamel and dentine in relationship to structural orientation of enamel prisms and dentinal tubules by optical coherence tomography (OCT). METHODS: Discs of 300-400µm thickness including enamel with cross-cut or long-cut prisms and mid-coronal dentine with cross-cut, long-cut or oblique-cut tubules were obtained from 28 non-carious freshly extracted human third molars sectioned at different directions. Each disc was placed over a metal plate and B-scan images were obtained by a swept-source OCT at 1330nm centre wavelength (Dental SS-OCT Prototype 1, Panasonic Healthcare Co., Ltd, Japan). Refractive index (n) calculated from optical path length and real thickness, and OCT signal slope (OCTSS) in relation to structural orientation were statistically compared among groups (α=0.05). The discs were sectioned and observed under SEM to confirm prism or tubule orientations. RESULTS: No significant differences were found in n and OCTSS among cross-cut and long-cut enamel groups (p>0.05). However, in dentine, there was a significant difference in both variables among different regions. In cross-cut tubule orientation, the average n and OCTSS values were significantly lower compared to other two groups (long-cut and oblique-cut) (p<0.05). There was a significant difference in both variables between enamel and dentine (p<0.05). CONCLUSIONS: Unlike enamel, refractive index and OCT signal patterns in dentine vary according to structural orientation, with dentine tubules playing the role. Attenuation of OCT signal intensity was small in enamel. The findings may contribute to a better understanding of the interactions of light with the dental tissue. CLINICAL SIGNIFICANCE: Precise records of refractive indices and OCT signal patterns may be important for clinical diagnosis of caries and measurement of structural depth for operative purposes using this technology. Effects of dentine structural orientation on refractive index and scattering pattern must be considered when observing human teeth by OCT cross-sectional imaging.

The effects of aging on shear bond strength and nanoleakage expression of an etch-and-rinse adhesive on human enamel and dentin.

PURPOSE: To evaluate the effects of increasing thermocycling regimens on bond strength and nanoleakage of an etch-and-rinse adhesive. MATERIALS AND METHODS: Adper Single Bond (3M ESPE) was used for bonding resin composite to human enamel and dentin. Specimens were stored in water (37°C, 24 h) and subjected to thermocycling (5°C and 55°C) in five groups: 1. control (no thermal cycling), 2. 100, 3. 500, 4. 2000, or 5. 10,000 cycles. The microshear bond test was performed at a crosshead speed of 1 mm/min (n = 16). Nanoleakage specimens were immersed in silver nitrate and FE-SEM micrographs were digitally analyzed to calculate silver penetration in three samples. RESULTS: The mean bond strength to enamel was not significantly affected by thermocycling. However, artificial aging decreased dentin bond strength in groups 4 and 5 compared to control. Weibull analysis indicated that the characteristic strength decreased gradually with aging in both substrates. All groups exhibited some nanoleakage, and a significant increase in silver penetration compared to the control was observed in enamel and dentin groups 2 to 5. CONCLUSION: A nanoleakage increase was detected in an earlier stage of aging than when a significant drop in bond strength was observed, with the dentin bond being more susceptible to deterioration.