Dentin Discs Research Articles

Effect of Application Deviations on Dentin Sealing of a Universal Adhesive: Permeability and Nanoleakage.

The objective of this study was to evaluate the effect that deviations from the recommended protocol of a universal adhesive system, applied to dentin according to the self-etch (SE) and the etch-and-rinse (ER) techniques, has on permeability and nanoleakage. Permeability: 60 extracted non-carious human third molars (N = 60) were sectioned to obtain 0.7-mm-thick dentin disks. The specimens were randomly assigned to three subgroups and treated with a universal adhesive system (Prime&Bond Active Universal) using the SE and ER techniques: (1) following the manufacturer's instructions with 5 seconds drying (MFR DRY 5S), (2) following the MFR DRY 10S, and (3) reduced application time of the adhesive to 5 seconds (APPL 5S). Nanoleakage: 12 additional 0.7-mm-thick dentin disks were prepared, treated and divided into six groups. They were immersed in 50 wt% ammoniacal silver nitrate and processed according to conventional methods for the analysis of nanoleakage under transmission electron microscopy. The results were statistically analyzed by two-way analysis of variance and post-hoc Bonferroni's test. Significant differences in permeability reduction were observed among the treatment groups (0.001). The results obtained for APPL 5S were significantly lower than the results obtained for both the MFR DRY 5S (p = 0.003) and MFR DRY 10S (p = 0.001). The reduced application time to 5 seconds creates imperfect dentin tubule sealing, which may explain clinical reports of postoperative sensitivity and early degradation of the resin-dentin interface.

Characterization of silver diamine fluoride cytotoxicity using microfluidic tooth-on-a-chip and gingival equivalents

ObjectiveThis study aims to characterize the cytotoxicity potential of silver diamine fluoride (SDF) on dental pulp stem cells (DPSC) and gingival equivalents. MethodsDPSC cultured on 96-well plates was exposed directly to SDF (0.0001–0.01%) and cell viability (IC50) quantified. Effect of SDF on DPSC viability under flow (with dentin barrier) conditions was evaluated using a custom-designed microfluidic "tooth-on-a-chip”. Permeability of dentin discs (0.5–1.5 mm thickness) was evaluated using lucifer yellow permeation assay. Dentin discs were treated with 38% SDF (up to 3 h), and cell viability (live/dead assay) of the DPSC cultured in the inlet (unexposed) and outlet (exposed) regions of the pulp channel was evaluated. To assess the mucosal corrosion potential, gingival equivalents were treated with 38% SDF for 3 or 60 min (OECD test guideline 431) and characterized by MTT assay and histomorphometric analysis. ResultsDPSC exposed directly to SDF showed a dose-dependent reduction in cell viability (IC50: 0.001%). Inlet channels (internal control) of the tooth-on-a-chip exposed to PBS and SDF-exposed dentin discs showed> 85% DPSC viability. In contrast, the outlet channels of SDF-exposed dentin discs showed a decreased viability of< 31% and 0% (1.5 and ≤1.0 mm thick dentin disc, respectively) (p < 0.01). The gingiva equivalents treated with SDF for 3 and 60 min demonstrated decreased epithelial integrity, loss of intercellular cohesion and corneal layer detachment with significant reduction in intact epithelial thickness (p < 0.05). SignificanceSDF penetrated the dentin (≤1 mm thick) inducing significant death of the pulp cells. SDF also disrupted gingival epithelial integrity resulting in mucosal corrosion.

Experimental pastes containing niobophosphate and 45S5 bioactive glasses for treatment of dentin hypersensitivity: dentin permeability and tubule obliteration.

This study tested the ability of bioactive pastes containing niobophosphate and 45S5 glasses to reduce dentin permeability and to obliterate dentinal tubules, as a mean of reducing human dentin hypersensitivity. Experimental pastes with concentrations of 10, 20, and 30 wt% of two bioactive glasses (45S5 or niobophosphate [NbG]) were formulated. A paste without bioactive glass (placebo) and a commercial paste (Nano P, FGM) were used as controls. Forty dentin disc specimens were obtained from caries-free extracted third human molars and divided in 8 groups (n = 5). Percentage of permeability (%Lp) was assessed in a dental permeability machine considering hydraulic conductance, immediately after pastes application and at day 7, day 14, and day 21. The precipitates formed on the surface of the dentin discs (and dentinal tubules) were analyzed by SEM/EDS and micro-Raman spectra. Data of dentin permeability (%) 2-way repeated-measures (ANOVA) and Holm-Sidak post-tests (α = 0.05). Dentinal tubule obliteration was visually (and elemental) evaluated and descriptively reported. The experimental bioactive glass pastes containing NbG and 45S5, regardless of the concentration, reduced dentin permeability in comparison with pastes without bioactive glasses (P < 0.05). The formulated placebo and commercial paste did not reduce permeability over time (P < 0.05). SEM/EDS and micro-Raman analyses showed that both type of bioactive pastes (NbG or 45S5-based) presented mineral precipitates obliterating the dentinal tubules at day 21. NbG seems to offer a better initial effect than 45S5, while at 21 days there is no difference between both glasses. Experimental bioactive pastes containing NbG and 45S5 (at concentrations of 10%, 20%, or 30%) have potential to reduce dentin permeability (over time) in comparison with pastes without bioactive glasses; and this occurs on behalf of obliteration of dentinal tubules by microparticle and precipitate formation. Bioactive pastes containing NbG and 45S5 may benefit patients presenting dentin hypersensitivity, because these pastes can start acting fast after application and maintain their action up to 21 days.

Assessment of the remineralisation induced by contemporary ion-releasing materials in mineral-depleted dentine.

Evaluate the ability of currention-releasing materials to remineralise bacteria-driven artificial caries lesions. Standardised class I cavities were obtained in 60 extracted human molars. Specimens underwent a microbiological cariogenic protocol (28days) to generate artificial caries lesions and then were randomly divided into four restorative groups: adhesive + composite (negative control); glass ionomer cement (GIC); calcium silicate cement (MTA); and resin-modified calcium silicate cement (RMTA). Microhardness analysis (ΔKHN) was performed on 40 specimens (10/group, t = 30days, 45days, 60days in artificial saliva, AS). Micro-CT scans were acquired (3/group, t = 0days, 30days, and 90days in AS). Confocal microscopy was employed for interfacial ultra-morphology analysis (2/group, t = 0days and 60days in AS). Additional specimens were prepared and processed for scanning electron microscopy (SEM) and FTIR (n = 3/group + control) to analyse the ability of the tested materials to induce apatite formation on totally demineralised dentine discs (60days in AS). Statistical analyses were performed with a significance level of 5%. Adhesive + composite specimens showed the lowest ΔKHN values and the presence of gaps at the interface when assessed through micro-CT even after storage in AS. Conversely, all the tested ion-releasing materials presented an increase in ΔKHN after storage (p < 0.05), while MTA best reduced the demineralised artificial carious lesions gap at the interface. MTA and RMTA also showed apatite deposition on totally demineralised dentine surfaces (SEM and FTIR). All tested ion-releasing materials expressed mineral precipitation in demineralised dentine. Additionally, calcium silicate-based materials induced apatite precipitation and hardness recovery of artificial carious dentine lesions over time. Currention-releasing materials can induce remineralisation of carious dentine. MTA shows enhanced ability of nucleation/precipitation of hydroxyapatite compared to RMTA and GIC, which may be more appropriate to recover severe mineral-depleted dentine.

Effect of three intracanal medicaments used in pulp regeneration on the push-out bond strength of mineral trioxide aggregate and calcium-enriched mixture: An in vitro study.

Background. The bond strength of the materials used as a cervical barrier in the pulp regeneration is essential for the success of treatment. This study aimed to evaluate the effects of triple antibiotic paste (TAP), double antibiotic paste (DAP), and simvastatin as intracanal medicaments on the dislodgement resistance of mineral trioxide aggregate (MTA) and calcium-enriched mixture (CEM). Methods. A total of 160 extracted human single-rooted teeth were selected, and root canal preparation was carried out. The teeth in each group were randomly divided into four subgroups: TAP, DAP, simvastatin, and the control group (without intracanal medicament). Four weeks after placing the medicaments, it was removed by sodium hypochlorite, and MTA and CEM were placed in the coronal third of the root canals. After a week, 2-mm-thick dentin disks were prepared from the coronal third of the roots, and the push-out test was performed using a universal testing machine. The data were analyzed using two-way ANOVA and independent t-test at a significance level of 0.05. Results. Regardless of the intracanal medicament, there was no significant difference between the overall bond strength of MTA (59.3±10 MPa) and CEM (55.8±11 MPa) (P=0.6). Furthermore, there were no significant differences in bond strength between the two intracanal medicament groups and the control group (P>0.05). Conclusion. Under the limitations of the current study, DAP, simvastatin, and TAP, as intracanal medicaments, did not adversely affect the push-out bond strength of CEM and MTA.

Attachment of human periodontal ligament fibroblasts to root dentin conditioned with different endodontic irrigants: An experimental study.

Background. Periradicular surgery is the last treatment option for teeth with persistent periradicular endodontic lesions. This study aimed to assess the adhesion of fibroblasts to root dentin conditioned with ethylenediaminetetraacetic acid (EDTA), MTAD, and QMix.Methods. Twelve dentin discs were fabricated of 6 human single-rooted teeth. Fibroblasts were isolated from the periodontal ligament (PDL) of a premolar tooth. The teeth were healthy and freshly extracted from the socket. The samples were divided into four groups for surface conditioning with (I) EDTA, (II) MTAD, (III) QMix, and the control group. Fibroblasts were cultured on conditioned dentin discs at 37°C, 95% air, and 5% CO2 for 4 hours and then rinsed with PBS three times to eliminate unattached cells from the surface. The mean counts of attached cells were calculated using a Neubauer chamber. Also, the attachment of fibroblasts was evaluated using scanning electron microscopy (SEM).Results. The mean counts of fibroblasts attached to root dentin in EDTA, QMix, MTAD, and control groups were 303±46, 243±41, 213±33, and 347±38, respectively. No significant difference was noted in the number of fibroblasts attached between MTAD, EDTA, and QMix and the control group (P>0.05). Under SEM, the fibroblasts were flat and spindle-shaped, with cytoplasmic processes covering the untreated dentin surface. In the experimental groups, the cells were rounder with fewer processes. All the three groups showed weaker adhesion to dentin compared to the control (untreated dentin) group.Conclusion. Under the limitations of this study, it was concluded that treating the dentin surface with EDTA, MTAD, or QMIX might not be an effective way to improve the adhesion of human PDL fibroblasts.

Smear layer deproteinization with NaOCl and HOCl: Do application/wash-out times affect dentin bonding of one-step self-etch adhesives?

This study investigated the influence of application/wash-out times of sodium hypochlorite (NaOCl) and hypochlorous acid (HOCl) on dentin bond strength of one-step self-etch adhesives (1-SEAs). Human coronal dentin discs with a standardized smear layer were pretreated with 6% NaOCl or 100 ppm HOCl for 5 s, 15 s, or 30 s, and washed out with water for 5 s, 15 s or 30 s with or without the application of Clearfil DC Activator (CDA). No pretreatment was used as a control. The discs were bonded with a 1-SEA (Bond Force II or Clearfil Universal Bond Quick) and microtensile bond strength (µTBS) was measured after 24 h. Pretreatment with NaOCl for 15 s and 30 s significantly decreased µTBS (p<0.05), irrespective of wash-out time. The application of CDA recovered µTBS but did not outperform the control group. Conversely, pretreatment with HOCl for 15 s and 30 s followed by 30 s wash-out time significantly increased µTBS of 1-SEAs (p<0.05), regardless of CDA application.

The influence of methodology on the comparison of cytotoxicity of total-etch and self-etch adhesive systems

ObjectivesThe present study aimed to compare the in vitro cytocompatibility of two etch-and-rinse (Adper Scothbond, Optibond) and two self-etch (Clearfill SE Bond and Single Bond Universal) dental adhesives through a dentin-barrier model with human pulp fibroblasts. MethodsHuman fibroblasts were placed on a plastic device containing 500μm human dentin discs treated with each adhesive or without treatment (control). Other groups were directly exposed to media conditioned with adhesive samples according to ISO 10993-5:2009. After 24h exposure, cell viability was assessed by XTT, and released inflammatory mediators were detected with a multiparametric immunoassay. ResultsThe standardized test without barrier indicated both etch-and-rinse adhesives and self-etch as cytotoxic, promoting viabilities under 70% of the control group (p<0.05). The dentin-barrier model identified increased cell viability for self-etch adhesives, with Clearfill SE Bond identified as non-cytotoxic. The immunoassay evidenced high rates of cytokines by cells exposed to the conditioned media of Adper Scotchbond, Optibond S, and Single Bond Universal. ConclusionsThe use of a dentin-barrier in vitro model detected a better biocompatibility for self-etching adhesives and, in the case of Clearfill SE Bond, with a reversion from cytotoxic to biocompatible when compared to the indirect standardized test. Clinical significanceThe use of a dentin-barrier in vitro model was able to detect a better biocompatibility for self-etching adhesives when compared to the indirect standardized test and presents itself as a predictive in vitro method for assessing the cytotoxicity of dental restorative materials that may simulate the clinical condition more accurately.

The response of dual-species bacterial biofilm to 2% and 5% NaOCl mixed with etidronic acid: A laboratory real-time evaluation using optical coherence tomography.

AimThe addition of etidronic acid (HEDP) to sodium hypochlorite (NaOCl) could increase the antibiofilm potency of the irrigant, whilst maintaining the benefits of continuous chelation. Studies conducted so far have shown that mixing HEDP with NaOCl solutions of relatively low concentration does not compromise the antibiofilm efficacy of the irrigant. However, the working lifespan of NaOCl may decrease resulting in a reduction of its antibiofilm efficacy over time (efficiency). In this regard, continuous irrigant replenishment needs to be examined. This study investigated the response of a dual‐species biofilm when challenged with 2% and 5% NaOCl mixed with HEDP for a prolonged timespan and under steady laminar flow.MethodologyDual‐species biofilms comprised of Streptococcus oralis J22 and Actinomyces naeslundii T14V‐J1 were grown on human dentine discs in a constant depth film fermenter (CDFF) for 96 h. Biofilms were treated with 2% and 5% NaOCl, alone or mixed with HEDP. Irrigants were applied under steady laminar flow for 8 min. Biofilm response was evaluated by means of optical coherence tomography (OCT). Biofilm removal, biofilm disruption, rate of biofilm loss and disruption as well as bubble formation were assessed. One‐way anova, Wilcoxon's signed‐rank test and Kruskal–Wallis H test were performed for statistical analysis of the data. The level of significance was set at a ≤.05.ResultsIncreasing NaOCl concentration resulted in increased biofilm removal and disruption, higher rate of biofilm loss and disruption and increased bubble formation. Mixing HEDP with NaOCl caused a delay in the antibiofilm action of the latter, without compromising its antibiofilm efficacy.ConclusionsNaOCl concentration dictates the biofilm response irrespective of the presence of HEDP. The addition of HEDP resulted in a delay in the antibiofilm action of NaOCl. This delay affects the efficiency, but not the efficacy of the irrigant over time.

Influence of ceramic veneer on the transdentinal cytotoxicity, degree of conversion and bond strength of light-cured resin cements to dentin

ObjectivesTo investigate the transdentinal cytotoxicity (TC), degree of conversion (DC), and micro shear bond strength (μSBS) to dentin of light-cured resin cements (LCRCs) photoactivated directly or through a ceramic veneer( ± CV). Materials and methodsThe TC was assessed using human dentin discs adapted into artificial pulp chambers. Odontoblast-like cells were seeded on the pulp surface of the discs, then three LCRCs( ± CV) were applied on their etched and hybridized occlusal surface (n = 8/group). The adhesive systems of each LCRCs and sterile phosphate-buffered saline were used as positive and negative controls, respectively. After 24 h, the viability and morphology of cells adhered on discs were assessed. The extracts (culture medium + components of the materials diffused through the discs) were applied on the MDPC-23 to evaluate their viability, adhesion/spreading (A/S), alkaline phosphatase activity (ALP), and mineralized nodule formation (MN). LCRCs( ± CV) specimens were evaluated concerning the DC and μSBS to dentin. Data were analyzed by one-, two-, or three-way ANOVA/Dunnett, Sidak, and Games-Howell tests (α = 5%). ResultsAll LCRCs( ± CV) reduced cell viability, A/S, ALP, MN, and DC. Except for μSBS, the intensity of reduction was dependent on the LCRC used. LCRCs+CV resulted in lower DC and μSBS but did not increase the TC. SignificanceBesides the presence of CV between the light source and LCRCs reduces the degree of conversion and bond strength to dentin, these materials cause variable level of transdentinal toxicity to pulp cells. Thus, the composition and curing protocols of LCRCs should be revisited and reinforced to prevent mechanical and biological drawbacks.

Application of Sulfinate Agent in Conjunction with HOCl Smear-Layer Deproteinization Improves Dentin Bonding Durability of One-step Self-etch Adhesives.

To evaluate the effect of a sulfinate agent on the bonding durability of one-step self-etch adhesives (1-SEAs) to smear-layer-covered dentin deproteinized with hypochlorous acid (HOCl). Human coronal dentin disks with a standardized smear layer were deproteinized with 100 ppm HOCl solution for 0 s (control), 15 s or 30 s. After rinsing with water for 30 s and air drying, half of the specimens were treated with a sulfinate agent (Scotchbond Universal Dual Cure Activator; SDA) prior to the application of a 1-SEA (Bond Force II [Tokuyama Dental] or Clearfil Universal Bond Quick [Kuraray Noritake]). Microtensile bond strength (µTBS) was measured after 24 h or 10,000 thermal cycles (TC). The data were analyzed by three-way ANOVA with Tukey's post-hoc tests and t-tests at the 0.05 significance level. The 24-h µTBS of both adhesives increased statistically significantly with the HOCl pretreatment for 15 s or 30 s (p < 0.05), but it was not statistically significantly affected by the application of SDA (p > 0.05). However, after TC, the groups treated with the combination of HOCl and SDA maintained their µTBS (p > 0.05), as opposed to untreated dentin and dentin treated with either HOCl or SDA, whose µTBS decreased significantly (p < 0.05). The application of the sulfinate agent did not statistically significantly affect the immediate bond strength of 1-SEAs, and it could not prevent a significant decrease in the bond strength to untreated dentin after thermocycling. However, the sulfinate agent significantly improved the bonding durability of 1-SEAs to HOCl smear-layer deproteinized dentin.

Effect of high-fluoride dentifrice on root dentine and bacterial composition in a multispecies biofilm model

The present study evaluated the effect of high-fluoride dentifrice on dentine demineralization and bacterial composition in a multispecies biofilm model in vitro. A seven-organism bacterial consortium was grown on bovine dentine discs in a high-throughput active attachment model. The biofilms were submitted twice per day to the following dentifrices treatments: 5,000 ppm F, 1,100 ppm F, with placebo as a negative control. After 5 days of biofilm growth, dentine samples were assessed by transversal microradiography, the biofilm was collected for bacterial counts and the pH of the media was determined. Lower integrated mineral loss values were observed when 5,000 ppm F-treatment was used compared to the other treatments. Overall microbiological counts decreased with increasing F-concentration as well the pH of the media throughout the experiment. The 5,000 ppm F-treatment caused a shift in microbial composition and reduced dentine demineralization in the in-vitro experimental model.

Effect of Diode Laser versus a Combination of Sodium Trimetaphosphate with Polyacrylic Acid on Obliteration of Dentinal Tubules: An In Vitro Study

BACKGROUND: Irradiation with diode laser and biomimetic remineralization are important methods in the treatment of hypersensitivity and dentin remineralization. AIM: The aim of the study was evaluation of the effect of diode laser versus the effect of sodium trimetaphosphate (STMP) with polyacrylic acid (PAA) on obliteration of dentinal tubules (DT). MATERIALS AND METHODS: Sixty dentin discs with a thickness of 2 mm were prepared and conditioned with EDTA for 15 s. Then, all dentin discs were divided into three main groups (20 discs each) according to the treatment method; control, diode laser treated, and STMP with PAA group (biomimetic group). Each group was subdivided into four subgroups (five discs each) according to the storage time; 2 h (To), 1 month (T1), 2 months (T2), and 3 months (T3), respectively. All samples were prepared to be analyzed after each time interval using environmental scanning electron microscope (ESEM). Comparison of differences of DT obliteration percentage made on each group before and after the treatment were performed using computer-assisted digital image analysis. RESULTS: Control group showed the least DT obliteration percentage, the samples of laser group recorded statistically significant increase in DT obliteration percentage at To in comparison to the biomimetic group. Meanwhile, at T1, there was no statistically significant difference between both laser and biomimetic groups. However, statistically significant decrease was recorded in laser group at T2 and T3, respectively. CONCLUSION: Irradiation with diode laser and biomimetic remineralization using PAA with STMP are a promising methods to obliterate opened DT effectively.

Promoting mineralization at biological interfaces Ex vivo with novel amelotin-based bio-nano complexes

Interfacial failure at the resin-dentin interface is a significant disadvantage of resin-based dental restoration. In this study, we created bio-inspired bio-nano complexes using the enamel protein amelotin (AMTN) or AMTN with an engineered collagen-binding site (AMTN-Col) to coat hydroxyapatite nanoparticles (HANP). The resulting nano-bio complexes, AMTN-HANP and AMTN-Col–HANP, were evaluated for their ability to promote collagen mineralization. Our study comprises three separate phases.In phase I, developing a method for functionalizing HANP with AMTN/AMTN-Col was explored. HANP were synthesized and characterized using TEM, SAED-TEM, XRD and ATR-FTIR. The nanoparticles were functionalized with AMTN or AMTN-Col. The successful coating of the nanoparticles with the proteins was confirmed using a TEM image of immunogold-labelled samples.In phase II of the study, the mineralization potential of the synthesized bio-nano complexes was studied using model systems consisting of simulated body fluid (SBF), polymerized collagen gels, and dentin disks prepared from human extracted molars. Mineral formation in SBF was recorded with a light scattering assay using a microplate reader on 8 replicates of each sample per study time point. Statistical analysis was performed using one-way ANOVA and the Tukey test. Significance was assigned at P ​< ​0.01. The extent of mineral formation on collagen gel and remineralization of demineralized dentin was studied with SEM. Accelerated mineral formation collagen mineralization of bio-nano complexes treated samples were observed in all model systems.In phase III of the study, the clinical utilization of AMTN/AMTN-Col coated HANP in bio-integration and enhancing the bond strength of a resin-based dental restoration and the dentin interface was investigated. The bio-nano complexes were applied as a pretreatment on dentin disks prepared from human extracted molars prior to the composite resin restoration. The micro-shear bond strength test was done on 8 samples per treatment group (a total of 32 samples). Statistical analysis on shear bond strength was performed using one-way ANOVA and the Tukey test. Significance was assigned at P ​< ​0.01. Shear bond strength values indicated that pretreatment of dentin with the bio-nano complexes before adhesive application significantly improved shear bond strength.ConclusionWe have shown that AMTN based bio-nano complexes promote mineral formation on collagenous interfaces. Our findings can be the basis of new bio-inspired, bio-nano materials that may improve dental restoration longevity by enhancing the stability and integrity of the dentin-composite resin interface.

Effect of hydroxyapatite based agents on the bonding properties of universal adhesives

Objective: To study the effect of hydroxyapatite (HA) based agents on the bonding properties of universal adhesive with different application modes, and to provide evidence for the use of adhesives after desensitization treatment. Methods: Sixty impacted third molars were extracted and selected (acquired from Department of Oral and Maxillofacial Surgery, College of Stomatology, Xi'an Jiaotong University). Four third molars were used to prepare 1 mm thick dentin disks and treated with 1% citric acid to simulate sensitive tooth models. The dentin surfaces were observed by scanning electron microscope (SEM) after treating with no desensitization (control group), desensitized by HA based toothpaste Biorepair and Dontodent Sensitive respectively (desensitizing toothpaste A group and B group), or HA paste treatment (desensitizing paste group ) (n=2 per group). The remaining teeth were selected to expose the mid-coronal dentin and establish dentin sensitivity models. Then, the specimens were divided into 4 former groups and received corresponding treatment. Each group was randomly divided into 2 subgroups, and intermediately strong universal adhesive (G-Premio Bond) was applied on the desensitized dentin by either etch-and-rinse mode or self-etch mode. Resin-dentin slice specimens (n=4 per subgroup), microtensile specimens (n=20 per subgroup) and slice specimens (n=6 per subgroup) were prepared. The microstructure and nanoleakage of the adhesive interfaces were observed by scanning electron microscopy (SEM). The microtensile strength (bond strength) and fracture mode were tested and recorded. The water permeability of the adhesive interface was observed by laser scanning confocal microscopy (LSCM). Results: SEM showed that desensitizing toothpaste and desensitizing paste could partially or entirely occlude most of the dentin tubules. For the etch-and-rinse mode, the bond strength of specimens treated with toothpaste A [(40.98±4.60) MPa], toothpaste B [(40.89±4.64) MPa] and HA paste [(41.48±3.65) MPa] was significantly higher than that of the control group [(38.58±4.28) MPa] (F=3.89,P<0.05). There was no statistically significant difference in bond strength among the 4 subgroups for self-etch modes (F=0.48,P>0.05). After desensitization, the bond strength of the control group and desensitizing groups in the self-etch mode was significantly higher than that in the etch-and-rinse mode (P<0.05). The overall fracture modes were mixed failure and interfacial failure in the control group and desensitizing groups. SEM showed speckled silver-stained particles deposited along the bottom of the hybrid layer on the bond interface of etch-and-rinse mode, and there were few silver-stained particles deposited on the bond interface of self-etch mode. LSCM showed continuous linear penetration in the hybrid layer of etch-and-rinse mode subgroups and discontinuous linear penetration in the hybrid layer of self-etch mode subgroups. Conclusions: HA based desensitizers have no adverse effect on the bond strength of intermediately strong universal adhesive and show good bonding performance accompanied with the self-etch mode.

Nanomechanical and Molecular Characterization of Aging in Dentinal Collagen

Methylglyoxal (MGO) is an important molecule derived from glucose metabolism with the capacity of attaching to collagen and generating advanced glycation end products (AGEs), which accumulate in tissues over time and are associated with aging and diseases. However, the accumulation of MGO-derived AGEs in dentin and their effect on the nanomechanical properties of dentinal collagen remain unknown. Thus, the aim of the present study was to quantify MGO-based AGEs in the organic matrix of human dentin as a function of age and associate these changes with alterations in the nanomechanical and ultrastructural properties of dentinal collagen. For this, 12 healthy teeth from <26-y-old and >50-y-old patients were collected and prepared to obtain crown and root dentin discs. Following demineralization, MGO-derived AGEs were quantified with a competitive ELISA. In addition, atomic force microscopy nanoindentation was utilized to measure changes in elastic modulus in peritubular and intertubular collagen fibrils. Finally, principal component analysis was carried out to determine aging profiles for crown and root dentin. Results showed an increased presence of MGO AGEs in the organic matrix of dentin in the >50-y-old specimens as compared with the <26-y-old specimens in crown and root. Furthermore, an increase in peritubular and intertubular collagen elasticity was observed in the >50-y-old group associated with ultrastructural changes in the organic matrix as determined by atomic force microscopy analysis. Furthermore, principal component analysis loading plots suggested different “aging profiles” in crown and root dentin, which could have important therapeutic implications in restorative and adhesive dentistry approaches. Overall, these results demonstrate that the organic matrix of human dentin undergoes aging-related changes due to MGO-derived AGEs with important changes in the nanomechanical behavior of collagen that may affect diagnostic and restorative procedures in older people.

Nanoindentation Mapping and Bond Strength Study of Adhesive–Dentin Interfaces

AbstractA scanning probe microscopy (SPM) nanoindentation technique is employed in this work to relate structure of adhesive–dentin interfacial regions with in situ mechanical properties. Noncarious human third molars are sectioned to obtain superficial, middle, and deep dentin disks. Each dentin disk is divided into two parts for bonding with All‐Bond Universal in etch‐and‐rinse (E&amp;R) and self‐etch (SE) modes. Nanoindentation arrays of a small‐scale region of 40 × 40 µm2 are configured, which include the whole adhesive interface area from the restorative composite across the adhesive interface to unaffected dentin. Micro‐shear bond strength (μSBS) is measured to evaluate the mechanical capability. By virtue of high‐resolution quantitative mapping of nano‐elastic modulus and nanohardness, the continuous hierarchical substructures of adhesive–dentin interfaces are visualized, including the composite, adhesive layer, hybrid layer, and unaffected dentin. A noticeable finding is the existence of a transition zone between the hybrid layer and unaffected dentin. The μSBS result indicates that mechanical interlocking between intertubular dentin and adhesives dominates the bond strength. SPM nanoindentation mapping can correlate surface structure with mechanical properties across adhesive interfaces. Even more, the present findings demonstrate that SPM‐mapping of combined structural and mechanical property data of adhesive interfaces can potentially be used to evaluate bonding performance.

In Vitro Effect of Instrumentation Using Ultrasonication with and without Hydrogen Peroxide on the Removal of Biofilms and Spread of Viable Microorganisms in Aerosols.

To evaluate the use of hydrogen peroxide as an adjunct to ultrasonication (US) in biofilm removal and whether it can limit the spread of viable microorganisms in the aerosol. Multi-species biofilms were formed on dentin disks and titanium disks fixed on a plastic surface. After placing the specimens in a periodontal pocket model, an ultrasonic scaler was applied for 30 s, in part combined with 0.25% or 0.5% H2O2. After treatment, the remaining biofilm was analysed for bacterial counts (colony forming units [CFU]), biofilm quantity and metabolic activity. Further, the cytotoxic effect of hydrogen peroxide on periodontal ligament fibroblasts was assessed and the spread of bacteria in aerosol was quantified. Ultrasonication reduced bacterial counts in biofilm, biofilm mass and metabolic activity on both dentin and titanium disks. Adjunctive use of 0.25% and 0.5% H2O2 more effectively reduced the viable bacteria in biofilm than ultrasonication alone; this was also found on both dentin and titanium. The different concentrations of H2O2 did not lead to corresponding differences in bacterial mass and metabolic activity. The spread of bacteria through aerosols was statistically significantly reduced when adjunctive H2O2 was used. However, a certain cytotoxic effect on periodontal ligament fibroblasts by H2O2 could not be ruled out. Irrigating with H2O2 during periodontal instrumentation with an ultrasonic scaler increases the reduction of viable bacteria within biofilms. It might limit bacterial spreading via aerosols.

Visualization of initial bacterial colonization on dentin using fluorescence activating headlight for fluorescence enhanced theragnosis

ObjectiveTo evaluate accuracy of caries detection and the application-sensitivity of the new Designs for Vision's REVEAL™ utilizing a fluorescence activating headlight for excitation purpose. Materials and methodsREVEAL dental fluorescence loupes and headlight system were used. Occlusal enamel was removed, and mid-coronal dentine was exposed. Carious artificial lesion was created. Streptococcus mutans, Actinomyces naeslundii, and Streptococcus sanguis were used. The assessment was performed using two diagnostic methods: naked eye and Design for Vision Glasses with inter examiner blinding using two calibrated examiners. After 7 days, Raman measurements were made on dentin disc specimens with 785 nm wavelength. The bacterial counts in colony-forming units (CFU) were used to examine the growth kinetics of biofilms. The collagen fibril structure within the discs was performed using Transmission Electron Microscope. Scanning Electron Microscope was used to image samples at various magnifications. FISH was performed with specimens fixed in 4% paraformaldehyde in phosphate-buffered saline. Reproducibility was measured by Cohen kappa scores, values of which range from 0 for less than chance agreement to 1 for almost perfect agreement (p < 0.05). ResultsA significant kappa score of 0.706 showing significant reproducibility for the given diagnostic techniques, as all the teeth included in the study were spotted with the lesions. Most bacteria were detected using the CFU technique. The Raman bands scanned across the dentin surface at 960 cm−1 (P–O peak) are assigned to hydroxyapatite phosphate vibrations. FISH identified nearly all stained bacteria as days and time and dental hard tissue had a significant impact on the number of adherent bacteria. Scanning electron micrographs of polished cross sections of demineralized and non-demineralized specimens with perpendicular each tubule orientation (zone of demineralized dentin inset. ConclusionFluorescent enhanced theragnosis through Reveal vision glasses can ensure constant monitoring and diagnosis of caries progress . This may allow for a better clinical outcome.

A novel in vitro periodontal pocket model to evaluate the effect of root surface instrumentation on biofilm-epithelial cell interactions.

ObjectiveTo develop a novel in vitro periodontal pocket model for evaluating the effect of two different root surface instrumentation modalities on biofilm-epithelial cell interactions.Materials and methodsAn artificial periodontal pocket model was created using an impression material. Dentin discs were prepared and incubated for 3.5 days with a biofilm consisting of 12 bacterial strains. Then, the discs were inserted into the pocket model and instrumented for 10 s or 10 strokes either with ultrasonics (US) or hand instruments (HI). Subsequently, a glass slide coated with epithelial cells was placed in close vicinity to the discs. After incubation of the pocket model in a 5% CO2 atmosphere for 6 h, residual bacteria of the biofilm as well as bacteria adhering to or invaded into epithelial cells were determined using colony-forming unit (cfu) counts and real-time PCR. Further, as a parameter of the pro-inflammatory cell response, interleukin (IL)-8 expression was determined by ELISA.ResultsCompared to untreated control, HI reduced the cfu counts by 0.63 log10 (not significant) and US by 1.78 log10 (p = 0.005) with a significant difference between the treatment modalities favoring US (p = 0.048). By trend, lower detection levels of Tannerella forsythia were detected in the US group compared to HI. Concerning the interaction with epithelial cells, half of the control and the HI samples showed epithelial cells with attaching or invading bacteria, while US displayed bacteria only in two out of eight samples. In addition, US resulted in significantly lower IL-8 secretion by epithelial cells compared to the untreated control. Between HI and controls, no statistically significant difference in IL-8 secretion was found.ConclusionThis newly developed in vitro model revealed in terms of biofilm-epithelial cell interaction after root surface instrumentation that compared to hand curettes, ultrasonic instrumentation appeared to be more effective in removing bacterial biofilm and in decreasing the inflammatory response of epithelium to biofilm.Clinical relevanceUltrasonic instrumentation might be more advantageous to reduce cellular inflammatory response than hand instruments.