Chemical, structural and cytotoxicity characterisation of experimental fluoride-doped calcium phosphates as promising remineralising materials for dental applications.

OBJECTIVES: This study aimed at evaluating the cytotoxicity, chemical and structural properties of experimental fluoride-doped calcium-phosphates as potential remineralising materials for dental applications. METHODS: Experimental calcium phosphates were formulated using ß-tricalcium phosphate, monocalcium phosphate monohydrate, calcium hydroxide, and different concentrations of calcium/sodium fluoride salts [(5 wt%: VSG5F), (10 wt%: VSG10F), (20 wt%: VSG20F)]. A fluoride-free calcium phosphate (VSG) was used as control. Each tested material was immersed in simulated body fluid (SBF), (24 h, 15 and 30 days) to assess their ability to crystallise into apatite-like. Cumulative fluoride release was assayed up to 45 days. Moreover, each powder was placed into a medium containing human dental pulp stem cells (200 mg/mL) and their cytotoxicity was analysed using the 3-(4,5-dimethylthiazol-2-yl)- 2,5-diphenyltetrazolium bromide (MTT) assay (24 h, 48 h and 72 h incubation). These latter results were statistically analysed by ANOVA and Tukey's test (α = 0.05). RESULTS: All the experimental VSG-F materials produced fluoride-containing apatite-like crystals after SBF immersion. VSG20F presented prolonged release of fluoride ions into the storage media (45d). VSG, VSG10F and VSG20F showed a significant cytotoxicity at dilution of 1:1, while at 1:5, only VSG and VSG20F demonstrated a reduction in cell viability. At lower dilutions (1:10, 1:50 and 1:100) all specimens showed no significant toxicity to hDPSCs, but an increase in cell proliferation. SIGNIFICANCE: The experimental fluoride-doped calcium-phosphates are biocompatible and possess a clear ability to evoke fluoride-containing apatite-like crystallisation. Hence, they may be promising remineralising materials for dental applications.

Evaluation of Foraminal Enlargement with Thermally Treated Nickel-Titanium Systems.

AIM AND OBJECTIVE: To evaluate morphologic changes in the main foramen after enlargement with four different systems. MATERIALS AND METHODS: One hundred and twenty canals (buccal of maxillary molars and mesial of mandibular molars) with patency and apical foramen ≤200 µm were included. These apical foramina were photographed and then randomly divided into four experimental groups, according to the ProDesign Logic (PDL), ProDesign R (PDR), twisted file adaptive (TFA), and WaveOne Gold (WOG) systems, all of which were composed of three sequential instruments (Glidepath, #25 and #35). Root canals were prepared 1 mm beyond the main foramen (n = 30). The specimens were photographed again to capture intermediate (#25) and final enlargement (#35) images. These were analyzed using an image software that allowed comparison of the enlarged area and shape of the main foramen. RESULTS: In the Intragroup comparison, the Friedman's test showed a statistical difference between each phase of foramen enlargement (p <0.05). In the intergroup comparison, the percentage of foramen enlargement showed statistically significant difference between Groups PDR and TFA only in the analysis from #25 to #35, with 42.88 and 67.34% of foramen area widening, respectively. Relative to the final foramen shape, 81.7% were observed to be circular, 17.5% oval, and 0.8% flattened. CONCLUSION: Our results showed that foramen enlargement allowed an increase in area, maintained the circular shape of the foramen in the majority of cases, irrespective of the system used, and was a feasible option for use during preparation of the root canal system.

Degradation of Adhesive-Dentin Interfaces Created Using Different Bonding Strategies after Five-year Simulated Pulpal Pressure.

PURPOSE: To compare after five-year simulated pulpal pressure (SPP) the degradation of adhesive-dentin interfaces created using two simplified adhesives applied with different bonding strategies. MATERIALS AND METHODS: A two-step self-etch (CSE: Clearfil SE Bond) adhesive was used as a control multistep adhesive. The tested experimental materials were two simplified adhesives, a one-step self-etch (CS3: Clearfil S3 Bond) and a self-priming etch-and-rinse adhesive (SB2: Adper Single-Bond 2). Half of the bonded specimens were submitted to microtensile bond strength (µTBS) testing after 24 h. The other half submitted to SPP for five years before µTBS testing. Nonfractured sticks were evaluated using transmission electron microscopy (TEM). Scanning electron microscopy (SEM) was used to evaluate silver-nitrate nanoleakage within the interface. Data were statistically analyzed by two-way ANOVA and Tukey's test (p < 0.05). RESULTS: Prolonged SPP induced bond-strength reduction for both SB2 and CS3. All bonding approaches showed increased nanoleakage after aging. The two simplified adhesives showed severe degradation at the resin-dentin interface. TEM revealed that the main degradation patterns for the etch-and-rinse adhesive SB2 was collagen breakdown, while polymer hydrolysis along with filler debonding was mainly observed in CS3. CONCLUSIONS: Simplified adhesives applied in etch-and-rinse mode are mainly characterized by hydrolysis and collagen degradation. In self-etch mode, simplified adhesives may principally show hydrolysis of the polymeric matrix and/or at the interface of fillers and coupling agent. The use of multistep self-etching adhesives may guarantee greater dentin bond durability compared to simplified adhesives.

Influence of nanogel additive hydrophilicity on dental adhesive mechanical performance and dentin bonding.

OBJECTIVE: To assess the influence of hydrophilicity of reactive nanogels on the mechanical performance of dental adhesives and microtensile bond strength (µTBS) to dentin after 24h or 3 months of aging. METHODS: A series of three nanogels were synthesized: NG1-IBMA/UDMA; NG2-HEMA/BisGMA; NG3-HEMA/TE-EGDMA. The nanogels were dispersed in solvent, HEMA or BisGMA/HEMA. The degree of conversion (DC) of the materials was measured and the flexural modulus of these polymers was evaluated in dry or wet conditions. For µTBS analysis, a model adhesive was used without nanogel (control) or with the incorporation of nanogels. µTBS was evaluated after storage in distilled water for 24h or 3 months. The analysis of the fracture was performed after µTBS testing. Data were analyzed using ANOVA and Tukey's test (α=0.05). RESULTS: Water significantly increased the modulus of NG1 and NG2 dispersed in solvent, while significantly decreased the stiffness of NG3. All polymers dispersed in HEMA and BisGMA/HEMA had significantly lower modulus when stored in water. NG2 showed the highest DC in solvent and BisGMA/HEMA. In HEMA, NG1 and NG3 produced the highest DC. After three months, NG2 showed the best µTBS. The µTBS of NG2-containing adhesive resin significantly increased after 3 months, while storage had no effect in the control group, NG1 and NG3. SIGNIFICANCE: The more hydrophobic IBMA/UDMA nanogel showed higher bulk material mechanical property results, but the best dentin bond strength values, and notably strength values that improved upon storage, were obtained with the amphiphilic nanogel based on BisGMA/HEMA.

Bonding performance of experimental bioactive/biomimetic self-etch adhesives doped with calcium-phosphate fillers and biomimetic analogs of phosphoproteins.

OBJECTIVES: This study examined the bonding performance and dentin remineralization potential of an experimental adhesive containing calcium-phosphate (Ca/P) micro-fillers, and self-etching primers doped with phosphoprotein biomimetic analogs (polyacrylic acid-(PAA) and/or sodium trimetaphosphate-(TMP)). METHODS: Experimental self-etching primers doped with biomimetic analogs (PAA and/or TMP), and an adhesive containing Ca(2+), PO4(-3)-releasing micro-fillers (Ca/P) were formulated. Sound human dentin specimens were bonded and cut into sticks after aging (24h or 6 months) under simulated pulpal pressure (20cm H2O), and tested for microtensile bond strength (µTBS). Results were analyzed using two-way ANOVA and Tukey's test (p<0.05). Interfacial silver nanoleakage was assessed using SEM. Remineralization of EDTA-demineralized dentin was assessed through FTIR and TEM ultrastructural analysis. RESULTS: Application of the Ca/P-doped adhesive with or without dentin pre-treatments with the primer containing both biomimetic analogs (PAA and TMP) promoted stable µTBS over 6 months. Conversely, µTBS of the control primer and filler-free adhesive significantly decreased after 6 months. Nanoleakage decreased within the resin-dentin interfaces created using the Ca/P-doped adhesives. EDTA-demineralized dentin specimens treated the Ca/P-doped adhesive and the primer containing PAA and TMP showed phosphate uptake (FTIR analysis), as well as deposition of needle-like crystallites at intrafibrillar level (TEM analysis). SIGNIFICANCE: The use of Ca/P-doped self-etching adhesives applied in combination with analogs of phosphoproteins provides durable resin-dentin bonds. This approach may represent a suitable bonding strategy for remineralization of intrafibrillar dentin collagen within the resin-dentin interface.

Advances in Dental Materials through Nanotechnology: Facts, Perspectives and Toxicological Aspects.

Nanotechnology is currently driving the dental materials industry to substantial growth, thus reflecting on improvements in materials available for oral prevention and treatment. The present review discusses new developments in nanotechnology applied to dentistry, focusing on the use of nanomaterials for improving the quality of oral care, the perspectives of research in this arena, and discussions on safety concerns regarding the use of dental nanomaterials. Details are provided on the cutting-edge properties (morphological, antibacterial, mechanical, fluorescence, antitumoral, and remineralization and regeneration potential) of polymeric, metallic and inorganic nano-based materials, as well as their use as nanocluster fillers, in nanocomposites, mouthwashes, medicines, and biomimetic dental materials. Nanotoxicological aspects, clinical applications, and perspectives for these nanomaterials are also discussed.

Effect of antioxidants on the dentin interface bond stability of adhesives exposed to hydrolytic degradation.

PURPOSE: This study assessed the effect of antioxidants vitamin C (Vit. C), vitamin E (Vit. E) and quercetin (Querc) on the dentin bonding performance, degree of conversion, and rate of polymerization of three commercial adhesive systems (Adper Single Bond 2 [SB], Clearfil SE Bond [CSE], Adper Easy Bond [EB]). MATERIALS AND METHODS: Human premolars were restored using antioxidant-doped adhesives. The samples were stored for 24 h in distilled water or 6 months under simulated pulpal pressure. Teeth were cut into sticks and the microtensile bond strength (µTBS) to dentin was tested in a universal testing machine. Qualitative nanoleakage analysis was performed from a central stick of each restored tooth. Degree of conversion and rate of polymerization of adhesive systems were evaluated in triplicate using real-time FT-IR. RESULTS: Although the inclusion of the antioxidants negatively affected the µTBS over 24 h, the antioxidant-doped adhesives maintained (SB-Vit. C, SB-Vit. E, CSE-Vit. C, EB-Querc) or increased (SB-Querc, CSE-Vit. E, CSE-Querc, EB-Vit. E, and EB-Vit. C) their µTBS during 6 months of storage. Only the µTBS of Adper Single Bond 2 dropped significantly after 6 months among the control groups. Slight changes in the nanoleakage pattern after aging were observed in all groups, except for the EB-control group, which showed a noteworthy increase in nanoleakage after 6 months, and for EB-Vit. C, which presented a remarkable decrease. A lower degree of conversion was obtained with all antioxidants in SB and EB, except for the EB-Vit. E group. Similar degrees of conversion were attained in control and experimental groups for CSE. The rate of polymerization was reduced in antioxidant-doped adhesives. CONCLUSION: The performance of antioxidants changed according to the adhesive system to which they were added, and antioxidant-doped adhesives appear to have a positive effect on the adhesive interface durability, since their bond strength obtained after 24 h was maintained or increased over time.

Methacrylate bonding to zirconia by in situ silica nanoparticle surface deposition.

OBJECTIVE: This study introduces an innovative method to enhance adhesion of methacrylate-based cements to yttria-stabilized zirconia (Y-TZP) by means of a silica-nanoparticle deposition process. METHODS: Two alkoxide organic precursors, tetraethyl-orthosilicate (TEOS) and zirconium tert-butoxide (ZTB) were diluted in hexane at different concentrations in order to obtain several experimental materials to enhance deposition of a SiO(x) reactive layer to Y-TZP. This deposition was attained via sintering alkoxide precursors directly on pre-sintered zirconia (infiltration method­INF) or application on the surface of fully sintered zirconia (coating method­COA). Untreated specimens and a commercial tribochemical silica coating were also tested as controls and all the treated Y-TZP specimens were analyzed using SEM-EDX. Specimens were bonded using silane, adhesive and dual-cure luting cement and submitted to shear bond strength test after different water storage periods (24 h, 3-, 6- and 12-months). RESULTS: SEM-EDX revealed Y-TZP surface covered by silica nanoclusters. The morphology of silica-covered Y-TZP surfaces was influenced by sintering method, employed to deposit nanoclusters. High bond strength (MPa) was observed when using COA method; highest TEOS percentage achieved the greatest bond strengths to Y-TZP surface (36.7±6.3 at 24 h). However, bonds stability was dependent on ZTB presence (32.9±9.7 at 3 months; 32.3±7.1 at 6 months). Regarding INF method, the highest and more stable resin-zirconia bond strength was attained when using experimental solutions containing TEOS and no ZTB. Both sintering methods tested in this study were able to achieve a bonding performance similar to that of classic tribochemical strategies. SIGNIFICANCE: This study demonstrates that it is possible to achieve a reliable and long-lasting bonding between yttria-stabilized zirconia ceramic and methacrylate-based cements when using this novel, simple, and cost-effective bonding approach.

Impact of hydrophilicity and length of spacer chains on the bonding of functional monomers.

OBJECTIVES: 10-Methacryloyloxy-decyl-dihydrogen-phosphate (10-MDP) is currently considered as one of the most effective functional monomers for dental bonding, this in part thanks to its long and relatively hydrophobic spacer chain, adequately separating the polymerizable from the phosphate functionalities. This study compared functional monomers with different spacer chains' length and hydrophilicity to 10-MDP on their dentin and enamel bonding performance. METHODS: Atomic absorption spectroscopy (AAS) was used to characterize the chemical interaction. Micro-tensile bond strength (µTBS) and fractographic analyses were performed after 24h and one year. Confocal micro-permeability and SEM nanoleakage assessments were also undertaken. The tested functional monomers were 2-MEP (2-carbon spacer), 10-MDP (10-carbon), 12-MDDP (12-carbon), MTEP (high hydrophilic polyether spacer chain) and CAP-P (intermediate hydrophilic ester spacer). RESULTS: AAS revealed clear differences (p<0.05) in monomer-calcium salt formation in this order: 12-MDDP=10-MDP>CAP-P>MTEP>2-MEP. The highest initial dentin µTBS was obtained using 10-MDP or 12-MDDP. After 1-year aging, a significant drop (p<0.05) in µTBS was observed for the adhesives with MTEP (enamel and dentin), 2-MEP (enamel) and CAP-P (enamel). MTEP presented the highest micro-permeability, while 2-MEP, CAP-P and MTEP showed increased nanoleakage after aging. SIGNIFICANCE: These outcomes showed that more hydrophilic and shorter spacer chains may compromise the chemical interaction with calcium and the dentin/enamel bonding performance.

The role of spacer carbon chain in acidic functional monomers on the physicochemical properties of self-etch dental adhesives.

OBJECTIVES: To evaluate the effects of acidic functional monomers with different hydrophilicity and spacer carbon chain length on the degree of conversion (DC), wettability (contact angle), water sorption (WS) and ultimate tensile strength (UTS) of experimental one-step self-etch adhesives (1-SEAs). METHODS: A series of standard resin blends was prepared with each formulation containing 15mol% of each acidic monomer. The structural variations of the acidic monomers were MEP (spacer chain with 2 carbons), MDP (10-carbons), MDDP (12-carbons), MTEP (more hydrophilic polyether spacer) and CAP-P (intermediate hydrophilicity ester spacer). Dumbbell-shaped and disc specimens were prepared and tested for UTS and WS, respectively. DC was assessed by FTIR, while the wettability of each 1-SEA was evaluated on glass slides and flat dentine surfaces. Results were analysed with one-way ANOVA and Tukey's test (p<0.05). RESULTS: The outcomes showed lower UTS for CAP-P, control blend and MEP than MTEP, MDDP and MDP (p<0.05). The degree of conversion was statistically similar for all resins (p=0.122). On dentine, the wettability was higher (lower contact angle) with the most hydrophilic monomer MTEP. Higher WS was attained using MTEP. Different lengths of the spacer chains did not result in different wettability and WS (p>0.05). CONCLUSION: At similar molar percentage, different acidic functional monomers induced similar degree of conversion and different UTS when included in a 1-SEA. However, the inclusion of highly hydrophilic monomer may increase the wettability on dentine and the WS.

Chemical interaction of 10-MDP (methacryloyloxi-decyl-dihydrogen-phosphate) in zinc-doped self-etch adhesives.

OBJECTIVES: Zinc-doped dentine adhesives have been recently advocated to interfere with metallo-proteinases-mediated collagen degradation. Nevertheless, there is a little information about the effects of zinc ions on the chemical interaction of self-etch functional monomers to dentine. The aim of this study was to assess if the inclusion of zinc into the primers of self-etch adhesives containing MDP (10-methacryloyloxi-decyl-dihydrogen-phosphate) may interfere with their chemical interaction to calcium/dentine. METHODS: Caries-free human molars were bonded using two commercial self-etching adhesives [Clearfil SE bond (CSE) and S3 bond (S3)] doped with zinc nitrate and submitted to microtensile bond strength (µTBS) and interface nanoleakage evaluation. Moreover, MDP was synthesised to evaluate the chemical interaction with calcium/dentine through atomic absorption spectroscopy (AAS) and SEM-EDX in the presence or absence of zinc ions. RESULTS: AAS showed increasing formation of MDP-zinc rather than MDP-calcium salts (p=0.002) in the presence of zinc. SEM-EDX confirmed the formation of zinc-rich phosphate deposits that were probably responsible for the significant reduction in µTBS and increased interfacial nanoleakage attained with zinc-doped CSE and S3. CONCLUSION: These outcomes demonstrated that the excessive presence of zinc ions may jeopardise the bonding performance of MDP-containing self-etch adhesives.

Evaluation of the micro-mechanical strength of resin bonded-dentin interfaces submitted to short-term degradation strategies.

The aim of this study was to evaluate the microtensile bond strength (µTBS) and confocal micropermeability of resin bonded-dentin specimens created using two representative two-step/self-etch adhesives submitted to short-term period degradation strategies such as simulated pulpal pressure, thermo- or mechanical-cycling challenges. Clearfil SE Bond (CSE) and Silorane adhesive (SIL) were bonded to flat deep dentin from seventy extracted human molars and light-cured for 10 s. Composite build-ups were constructed using with Filtek Z350 XT and Filtek P90 respectively. The specimens of each adhesive group were subjected to three different accelerated aging methods: (1) thermo-cycling challenge (5000 cycles); (2) mechanical-cycling load (200,000 cycles); (3) experiment and (4) conventional method for simulated pulpal pressure (20 cm H2O). Control resin-bonded specimens were stored in distilled water for 24 h. µTBS and confocal microscopy (CLSM) micropermeability evaluation were performed and the results were analyzed using Two-way ANOVA and Tukey's tests (α=0.05). The CLSM evaluation revealed micro-cracks within the Silorane-bonded dentin subsequent to mechanical-cycling load, whereas, the simulated pulpal pressure induced evident micropermeability in both bonding agents. Mechanical loading provides discernible bonding degradation in a short-term period in resin-bonded dentin created using two-step/self-etch adhesives. However, simulated pulpal pressure may reduce the sealing ability of self-etch adhesives causing greater water uptake within the resin-dentin interface.

Hydrolytic degradation of the resin-dentine interface induced by the simulated pulpal pressure, direct and indirect water ageing.

OBJECTIVES: The aim of this study was to compare the hydrolytic effects induced by simulated pulpal pressure, direct or indirect water exposure within the resin-dentine interfaces created with three "simplified" resin bonding systems (RBSs). METHODS: A two-step/self-etching (CSE: Clearfil SE Bond), one-step/self-etching (S3: Clearfil S3) and etch-and-rinse/self-priming (SB: Single-bond 2) adhesives were applied onto dentine and submitted to three different prolonged (6 or 12 months) ageing strategies: (i) Simulated Pulpal Pressure (SPP); (ii) Indirect Water Exposure (IWE: intact bonded-teeth); (iii) Direct Water Exposure (DWE: resin-dentine sticks). Control and aged specimens were submitted to microtensile bond strength (µTBS) and nanoleakage evaluation. Water sorption (WS) survey was also performed on resin disks. Results were analysed with two-way ANOVA and Tukey's test (p < 0.05). RESULTS: The µTBS of CS3 and SB dropped significantly (p < 0.05) after 6 months of SPP and DWE. CSE showed a significant µTBS reduction only after 12 months of DWE (p = 0.038). IWE promoted no statistical change in µTBS (p > 0.05) and no evident change in nanoleakage. Conversely, SPP induced a clear formation of "water-trees" in CS3 and SB. WS outcomes were CS3 > SB = CSE. CONCLUSION: The hydrolytic degradation of resin-dentine interfaces depend upon the type of the in vitro ageing strategy employed in the experimental design. Direct water exposure remains the quickest method to age the resin-dentine bonds. However, the use of SPP may better simulate the in vivo scenario. However, the application of a separate hydrophobic solvent-free adhesive layer may reduce the hydrolytic degradation and increase the longevity of resin-dentine interfaces created with simplified adhesives.