Toxicity and cytokine release from human dental pulp stem cells after exposure to universal dental adhesives cured by single peak and polywave LEDs.

OBJECTIVES: to assess the impact of universal adhesives, cured with single-peak and polywave LEDs, on the metabolic activity and cytokine release of human dental pulp stem cells (hDPSCs). In addition, analyze the degree of conversion (DC) of the adhesives cured with the different LEDs. METHODS: Discs (5 mm diameter, 1 mm thick) were prepared using three universal adhesives: Single Bond Universal (SBU, 3 M ESPE), Optibond Universal (OBU, Kerr), and Zipbond Universal (ZBU, SDI). These discs were cured for 40 s using a single-peak (DeepCure, 3 M ESPE) or a polywave light-emmiting diode (LED) curing unit (Valo Grand, Ultradent). After 24 h, the specimens were placed in 24-well culture plates, each containing 1 mL of culture medium for 24 h. hDPSCs (1.8 ×104) were seeded in 96-well plates and allowed to grow for 24 h. Subsequently, the cells were exposed to the extracts (culture medium containing eluates from the adhesive discs) for an additional 24 h. Cells not exposed to the extracts were used as a control group. The mitochondrial metabolism was assessed using the MTT assay and the cytokine release evaluated through MAGPIX. The degree of conversion of the adhesives was analyzed using FTIR (n = 5). The results were analyzed by ANOVA two-way and Tukey's test. RESULTS: OBU and ZBU eluates caused a statistically significant reduction in mitochondrial metabolism, regardless of the LED used, indicating their cytotoxicity. In contrast, SBU did not significantly affect the MTT results, resembling the control group. A higher release of cytokines IL-1, IL-6, IL-10, and TNF-α were found in association to ZBU. SBU, on the other hand, increased the release of IL-8. OBU did not influenced the cytokine release. SBU presented the higher DC, while OBU and ZBU had similar DC, lower than SBU. SIGNIFICANCE: In conclusion, universal adhesives exhibit toxicity towards hDPSCs, but the extent of toxicity varies depending on the adhesive material. ZBU was associated with increased cytokine release, particularly pro-inflammatory mediators, from hDPSCs. The different LEDs did not influenced the cytotoxicity of the evaluated adhesives.

Interaction of different concentrations of 10-MDP and GPDM on the zirconia bonding.

OBJECTIVES: To analyze the influence of different concentrations of 10-MDP and GPDM used, combined or not, on the bonding to zirconia. METHODS: Specimens of zirconia and a resin-composite (7 mm length, 1 mm width, and 1 mm thick) were obtained. The experimental groups were obtained according to the functional monomer (10-MDP and GPDM) and the concentrations (3 %, 5 %, and 8 %). For the groups with combined 10-MDP and GPDM, the agents were used with a proportion of 50 %/50 % wt until reaching the 3 %, 5 % and 8 % concentrations. All monomers were diluted in ethanol to obtain the primers. Two control groups were established: ethanol (negative control) and a commercial reference Monobond N (positive control). The zirconia surface treatment was performed with the primer application followed by the bonding to a resin-composite sample with a light-curing resin cement. Twenty-four hours after the adhesive procedure, a microtensile test was performed and the failure pattern of each sample was analysed with a stereoscopic magnifying glass. Data were analysed by a two-way ANOVA and Dunnet test. RESULTS: All experimental primers presented a higher bond strength than the negative control (ethanol). Excepting the 8 % GPDM primer, all groups presented statistically similar bond strength compared to the positive control, with a predominance of adhesive failure. SIGNIFICANCE: 10-MDP, GPDM, and the combination of both for the concentrations tested promote an effective chemical bonding to zirconia. However, using 10-MDP and GPDM in the same primer has no synergistic effect.

Co-initiators of polymerization can modulate the inflammatory cytokine release without major cytotoxic effects in human dental pulp cells.

To evaluate the cytotoxicity of co-initiators of polymerization and its influence on cytokine release from human dental pulp cells (hDPCs). Cells were isolated from the dental pulp of sound human third molars. The co-initiators dimethylaminoethyl amine benzoate-(EDAB), 2-(dimethylamino)ethyl methacrylate (DMAEMA); 2-Ethylhexyl 4-(dimethylamino)benzoate (EHA) and bis(4-methyl phenyl)iodonium hexafluorophosphate (BPI) were diluted in dimethylsulfoxide (DMSO) at different concentrations. In this way, experimental groups and one control (without treatment) were obtained. hDPCs (10 × 104 cell per well) were seeded on 96 well plates and incubated at 37°C and 5% CO2 for 48 h. After this, the cells were exposed to different concentrations of co-initiators cited for 24 h. After this time, the culture medium was removed, and the mitochondrial metabolism was evaluated by MTT assay, cell death by flow cytometry, and cytokine released (IL-1ß, IL6, IL-8, IL-10, and TNF-α) was analyzed by MAGPIX assay. The data were analyzed by ANOVA one-way and Tukey's test. EHA, DMAEMA, and EDAB did not reduce the mitochondrial metabolism. BPI presented high toxicity with remarkable reduction (80%) after exposure to 1 mM. The cell death of all test groups was similar to control. After 24 h treatment, the IL-8 was up-regulated by all compounds, while IL-6 was upregulated after exposure to EHA and downregulated after DMAEMA stimulation. BPI, EHA, EDAB, and DMAEMA can trigger an initial inflammatory response, upregulating the IL-8 secretion in hDPCs in a compound-concentration-dependent manner; however, this was not accompanied by major cytotoxic effects at cell death or mitochondrial-metabolism levels.

RoBDEMAT: A risk of bias tool and guideline to support reporting of pre-clinical dental materials research and assessment of systematic reviews.

OBJECTIVES: To develop a risk of bias tool for pre-clinical dental materials research studies that aims to support reporting of future investigations and improve assessment in systematic reviews. METHODS: A four-stage process following EQUATOR network recommendations was followed, which included project launch, literature review, Delphi process and the tool finalization. With the support of the European Federation of Conservative Dentistry (EFCD) and the Dental Materials Group of the International Association for Dental Research (DMG-IADR), a total of 26 expert stakeholders were included in the development and Delphi vote of the initial proposal. The proposal was built using data gathered from the literature review stage. During this stage, recent systematic reviews featuring dental materials research, and risk of bias tools found in the literature were comprehensively scanned for bias sources. The experts thus reached a consensus for the items, domains and judgement related to the tool, allowing a detailed guide for each item and corresponding signalling questions. RESULTS: The tool features nine items in total, spread between 4 domains, pertaining to the following types of bias: bias related to planning and allocation (D1), specimen preparation (D2), outcome assessment (D3) and data treatment and outcome reporting (D4). RoBDEMAT, as presented, features signalling questions and a guide that can be used for RoB judgement. Its use as a checklist is preferred over a final summary score. CONCLUSION: RoBDEMAT is the first risk of bias tool for pre-clinical dental materials research, supported and developed by a broad group of expert stakeholders in the field, validating its future use. CLINICAL SIGNIFICANCE: This new tool will contribute the study field by improving the scientific quality and rigour of dental materials research studies and their systematic reviews. Such studies are the foundation and support of future clinical research and evidence-based decisions.

Influence of different concentrations of an iodonium salt on properties of amine-free resins.

OBJECTIVES: to evaluate the effect of different concentrations of an iodonium salt on the degree of conversion, polymerisation rate, flexural strength and modulus of an amine-free photopolymerizable resin system. METHODS: Two series of UDMA/TEGDMA resin mixtures (1:1 mass ratio) containing 0, 0.5, 1.0 or 2.0 mol% of bis(4-methyl phenyl)iodonium hexafluorophosphate(BPI): and 1 mol% camphorquinone (CQ) were evaluated. One of the series contained 2 mol% of the amine (2-(dimethylamino)ethyl methacrylate-DMAEMA), while the other series had only CQ and BPI as photoinitiators. Polymerisation kinetics and degree of conversion (DC) were evaluated by Fourier transform near-infrared spectroscopy (n = 3). Flexural strength (FS) and modulus (E) were evaluated by a three-point bending method (n = 12). Data were analysed by two-way ANOVA and Tukey's test. RESULTS: The system containing CQ + DMAEMA + BPI provided the highest DC and FS. Amine-free resins had similar E regardless of the presence of BPI. The resins containing CQ+DMAEMA with 1 and 2 mol% of BPI presented higher E compared with others. SIGNIFICANCE: Amine-containing resins reached statistically higher DC, rate of polymerisation and FS than amine-free systems. The use of higher BPI concentrations did not improve the properties of amine-free resins.

Improvement on properties of experimental resin cements containing an iodonium salt cured under challenging polymerization conditions.

OBJECTIVE: The use of resin cements in clinical practice entails photopolymerization through prosthetic devices, which precludes light penetration. The objective of this study was to modify experimental resin cements (ERCs) with diphenyliodonium hexafluorophosphate (DPI) in an attempt to improve chemical and mechanical properties of materials cured with reduced irradiance and final radiant exposure. METHODS: A co-monomer base containing a 1:1 mass ratio of 2.2-bis[4-(2-hydroxy-3-methacryloxypropoxy)phenyl]propane (bis-GMA) and triethyleneglycol dimethacrylate (TEGDMA) was prepared, with 1mol% of camphorquinone and 2mol% of ethyl 4-(dimethylamino)benzoate as initiator system. The resin was divided into 4 fractions according to the DPI concentrations (0, 0.5, 1 and 2mol%). The challenging polymerization condition was simulated performing the light activation (12, 23 and 46s) through a ceramic block (3mm thick). The irradiance was assessed with a calibrated spectrometer (1320mW/cm2), resulting in three levels of radiant exposure (0.58, 1.1 and 2.2J/cm2). The polymerization kinetics was evaluated in real-time using a spectrometer (Near-IR). Water sorption and solubility was analyzed and the cohesive strength of resins obtained through the microtensile test. Polymerization stress was assessed by Bioman method. RESULTS: Resins containing DPI had higher degree of conversion and rate of polymerization than the control (without DPI). The use of DPI reduced water sorption and solubility, and led to higher cohesive strength compared to resins without the iodonium salt. However, the stress of polymerization was higher for experimental resins with DPI. SIGNIFICANCE: Even under remarkably reduced irradiance, cements containing a ternary initiating system with an iodonium salt can present an optimal degree of conversion and chemical/mechanical properties.

Physicochemical properties of dental resins formulated with amine-free photoinitiation systems.

OBJECTIVE: To assess the mechanical properties of two different dimethacrylate resin blends containing the photosensitizer camphorquinone (CQ) alone or in combination with one or more synergists including an amine and/or an iodonium. METHODS: Two co-monomer resin blends were formulated using Bis-GMA/TEGDMA and UDMA/TEGDMA, both at 1:1 mass ratio. Each resin blend was divided into four groups, comprising the following four photoinitiation systems: (1) CQ + 2-(dimethylamino)ethyl methacrylate (DMAEMA); (2) CQ + DMAEMA + bis(4-methyl phenyl)iodonium hexafluorophosphate (BPI); (3) CQ; and (4) CQ + BPI. Materials were evaluated for polymerisation kinetics, water sorption, solubility, flexural strength and modulus. RESULTS: BisGMA/TEGDMA with CQ showed minimal and insignificant degree of conversion and was not tested for water sorption/solubility and mechanical properties. The ternary system (i.e., CQ + DMAEMA + BPI), promoted the highest degree of conversion for each monomer blend. The resins containing amine had higher mechanical properties than the amine free. However, the UDMA amine free resins exhibited greater flexural strength and modulus than the corresponding amine free BisGMA resins. BisGMA/TEGDMA containing CQ + DMAEMA or CQ + BPI had significantly higher water sorption and solubility than the other groups. SIGNIFICANCE: Resins containing amine presented better properties than the amine-free systems. The addition of iodonium salt (BPI) improved the degree of conversion of the resins, even without an amine co-initiator. The amine-free initiator system (CQ + BPI) was more effective when used with UDMA versus BisGMA based-resins respectively.

Positive Synergistic Effect of Adhesives and Resin Cements Containing an Iodonium Salt on Bonding of Fiber Posts.

PURPOSE: To evaluate the push-out bond strength (PBS) of glass-fiber posts (GFP) in different root canal zones (cervical, middle, and apical), bonded with experimental simplified adhesives (ESAs) containing different initiator systems (camphorquinone [CQ] or phenylpropanodione [PPD]) with or without diphenyl iodonium hexafluorophosphate (DPI), in combination with a DPI-containing composite cement. MATERIALS AND METHODS: ESA blends were prepared with bisphenol glycidyl methacrylate (bis-GMA), triethylene glycol dimethacrylate (TEG-DMA), 1,3-glycerol dimethacrylate (GDMA), 2-hydroxyethyl methacrylate (HEMA), and ethanol, then divided into 12 experimental groups (n = 10) according to the initiator systems (CQ, PPD, or CQ + PPD) and the presence or absence of DPI. The roots of 120 extracted bovine incisors were prepared with #5 Largo drills and the GFP were cemented with each ESA and experimental composite cements containing 0.05 mol% of DPI. The push-out bond strength (PBS) test was performed after 24 h of storage. Failure patterns were analyzed under a stereomicroscope. Data were analyzed with split-plot two-way ANOVA and Tukey's test (α = 0.05). RESULTS: PBS was significantly higher for DPI-containing ESAs in all regions evaluated, with the group containing 0.5 CQ + 1 PPD + 0.5 DPI exhibiting the highest PBS. There was no statistically signficant difference among groups without DPI. Most failures were classified as adhesive at the cement-dentin interface. CONCLUSIONS: The combination of an adhesive and a composite cement containing DPI salt can improve GFP bonding to root dentin, even in the apical region.

Transmission of violet and blue light and current light units through glass-reinforced ceramics with different thicknesses.

Purpose To evaluate the effect of glass-reinforced ceramics (leucite and lithium disilicate) with different thicknesses (1, 2, and 3 mm) on the wavelength and irradiance spectrum of blue and violet lights. In addition, the effect of the ceramics on four current light-curing units (LCUs) was evaluated: a halogen lamp, a single peak LED, and two multi-peak LEDs.Methods Ceramic discs of different thicknesses (1, 2, and 3 mm) were obtained from computer-aided design and computer-aided manufacturing (CAD-CAM) blocks. The irradiance, radiant exposure, and emission spectrum of the four LCUs were analyzed using a spectrometer-based instrument. To evaluate the violet and blue lights, a specific device that provides a narrow emission spectrum was used.Results The ceramics reduced the irradiance of all the tested LCUs. However, the wavelength of the transmitted light was only altered slightly. The effect of leucite and lithium disilicate varied according to the type of LCU and thickness of the ceramic disc evaluated .Conclusions From the results, it could be concluded that the thickness of the leucite and lithium disilicate ceramic significantly reduced the irradiance of the light emitted by the LCUs, with minimal changes on the wavelength spectrum of the lights. The effects of the ceramic on irradiance and transmitted wavelengths of the blue and violet lights was slightly different.

Viscosity and thermal kinetics of 10 preheated restorative resin composites and effect of ultrasound energy on film thickness.

OBJECTIVE: This study investigated viscosity and thermal kinetics of 10 selected preheated restorative resin composites and the effect of ultrasound energy on film thickness. METHODS: A range of different resin composites was tested: Charisma Diamond, IPS Empress Direct, Enamel Plus HRi, Essentia, Estelite Omega, Filtek Z100, Filtek Z350 XT, Gradia, TPH Spectrum and VisCalor. A flowable resin composite (Opallis Flow) and two resin cements (RelyX Veneer, Variolink Esthetic LC) also were tested. Viscosity (Pa s) was measured at 37 °C and 69 °C (preheating temperature) using a rheometer. Film thickness (µm) was measured before and after application of ultrasound energy. Temperature loss within resin composite following preheating (°C/s) was monitored. Data were statistically analyzed (α = 0.05). RESULTS: Viscosity at 69 °C was lower than at 37 °C for all materials except the flowable resin composite. Preheating reduced viscosity between 47% and 92% for the restorative resin composites, which were generally more viscous than the flowable materials. Film thickness varied largely among materials. All preheated resin composites had films thicker than 50 µm without ultrasound energy. Application of ultrasound reduced film thickness between 21% and 49%. Linear and nonlinear regressions did not identify any relationship between filler loading, viscosity, and/or film thickness. All materials showed quick temperature reduction following preheating, showing maximum temperature loss rates after approximately 10 s. SIGNIFICANCE: Distinct restorative resin composites react differently to preheating, affecting viscosity and film thickness. The overall performance of the preheating technique depends on proper material selection and use of ultrasound energy for reducing film thickness.

Solvent and HEMA Increase Adhesive Toxicity and Cytokine Release from Dental Pulp Cells.

The aim of the present study was to evaluate the effect of the hydroxyethyl-methacrylate (HEMA) concentration and solvent content of dental adhesives on cell viability and cytokine (IL-1b, IL-6, IL-10, TNF-α) release by human dental pulp cells (HDPCs). HDPCs were obtained from fresh extracted human third molars. Experimental adhesives were prepared containing different concentrations of HEMA (0%, 10%, and 20%) with and without solvent (ethanol 10%). Cylindrical specimens were immersed on culture medium during 24 h to obtain the extracts. The cells were incubated with extracts (culture medium + components leached from the adhesives) of different adhesives, and cell viability and cytokine release were evaluated after 6 and 24 h of exposure. Adhesives containing HEMA promoted high cell viability reduction after 6 h of exposure; but after 24 h, the results were similar to the ones found among control group cells. These effects on cell viability were prominently increased with the addition of solvent. Although IL-1b release was not affected by exposure to eluates, other cytokines (IL-10, IL-6, TNF-α) were modulated by the different experiment conditions, directly influenced by the HEMA concentration and presence of solvent. Higher HEMA concentrations, combined with the presence of solvent, can promote significant reduction on HDPC viability, increasing the release of anti- and pro-inflammatory mediators.

Effects of Laser Irradiation on Pulp Cells Exposed to Bleaching Agents.

The aim of this study was to evaluate the effect of low-level laser therapy (LLLT) on odontoblast-like cells exposed to a bleaching agent. Mouse dental papilla cell-23 cells were seeded in wells of 24-well plates. Eight groups were established according to the exposure to the bleaching agent and LLLT (0, 4, 10 and 15 J cm(-2) ). Enamel-dentin disks were adapted to artificial pulp chambers, which were individually placed in wells containing Dulbecco's modified Eagle's medium (DMEM). A bleaching agent (35% hydrogen peroxide [BA35%HP]) was applied on enamel (15 min) to obtain the extracts (DMEM + BA35%HP components diffused through enamel/dentin disks). The extracts were applied (1 h) to the cells, and then subjected to LLLT. Cell viability (Methyl tetrazolium assay), alkaline phosphatase (ALP) activity, as well as gene expression of ALP, fibronectin (FN) and type I collagen, were evaluated. The bleaching procedures reduced the cell viability, ALP activity and gene expression of dentin proteins. Laser irradiation did not modulate the cell response; except for FN, as LLLT decreased the gene expression of this protein by the cells exposed to the BA35%HP. It can be concluded that BA35%HP decreased the activities of odontoblasts that were not recovered by the irradiation of the damaged cells with low-level laser parameters tested.

Effect of increased dwell times for solvent evaporation on the bond strength and degree of conversion of an ethanol-based adhesive system.

This study evaluated the influence of the prolonged setting time of an ethanol-based adhesive system on the dentin bond strength and degree of conversion. Labial and lingual surfaces of fifteen human third molars were flattened until the dentin was exposed and randomly allocated to 3 groups (n = 10), according to the dwell time between the application of two consecutive layers of the adhesive system (Adper Single Bond Plus, 3M ESPE) and light activation: G1--control (no extra dwell time); G2 and G3--dwell time of 30 seconds and 60 seconds, respectively. After light curing, two cylinders (1.4 x 1 mm) of composite resin (Filtek Flow, 3M ESPE) were bonded to each surface and submitted to micro-shear testing, 24 hours after light curing. A similar adhesive procedure was used for the degree of conversion evaluation using Fourier transform infrared spectroscopy (FTIR). Significant differences between bond strength values (p = 0.0003) and degrees of conversion (p = 0.0004) were detected. The bond strength of G3 (60-second dwell time) was statistically higher than that of other groups. G1 (control) and G2 (30-second dwell time) presented similar results. Values of degree of conversion indicated that both the 30-second and 60-second dwell times resulted in similar and greater percentages of conversion. The use of a longer dwell time (60 seconds) might provide better solvent volatilization and monomer infiltration; bringing benefits to dentin bonding using simplified etch & rinse adhesive systems.

Degree of conversion of etch-and-rinse and self-etch adhesives light-cured using QTH or LED.

In the current study, the degree of conversion (DC) of bonding agents photoactivated using QTH or LED light-curing units (LCUs) was evaluated by Fourier Transform infrared spectroscopy with an attenuated total reflectance (ATR) device. Four LCUs were evaluated: one QTH (Optilux 501; Demetron Kerr) and three LEDs: Radii Cal (SDI), Elipar FreeLight 2 (3M ESPE) and Bluephase (Ivoclar Vivadent). Two etch-and-rinse (Scotchbond Multi-Purpose-SBMP and Single Bond 2-SB2) and two self-etch adhesives (Clearfil SE Bond-CSE, and Clearfil S3 Bond-CS3) were tested. For SBMP and CSE, the primer was not used. The irradiance and spectral emission of the LCUs were obtained with a radiometer and spectrometer. The materials were placed onto the ATR cell as thin films, the solvent was evaporated (when necessary) and photoactivation was carried out for 20 seconds. The DC (%) was evaluated after five minutes (n = 5). The data were statistically analyzed (p < 0.05). The irradiance for Optilux, Radii, FreeLight 2 and Bluephase was 760, 600, 1000 and 1100 mW.cm(-2), respectively. The wavelength of emission for Optilux was between 375 and 520 nm (peak at 496 nm), while for Radii, it was between 420 and 520 nm (peak at 467 nm). Freelight 2 presented an emission spectrum between 415 and 520 nm, and for Bluephase, it was between 410 and 530 nm, both having a peak at 454 nm. SB2 generally showed higher DC compared with the other bonding agents. When cured using the QTH unit, the DC results were SB2 = CS3 > CSE > SBMP; for all LEDs, the DC results showed SB2 > CSE > SBMP > CS3. For SB2, the highest DC was observed when the material was cured with Radii, while there were no significant differences among the other LCUs. CSE and CS3 showed higher DC when cured using the QTH unit, but similar results were observed among the LEDs. For SBMP, no significant differences among the LCUs were detected. In conclusion, the combination bonding agent vs curing unit had a significant effect on DC, mainly for the self-etch adhesives.

Influence of retentive areas associated to onlay preparations on the dimensional stability of silicone impression materials.

This study evaluated the effect of retentive areas on onlay preparations on the dimensional alterations in condensation and addition silicone materials. A standard model with an onlay preparation was made. Each impression material was used through the double or simultaneous impression technique (n=25), resulting in a hundred impressions of the same model. Impressions were poured with type IV dental stone. Digital images were taken with a light microscope and the distances between the reference points created on the plaster dies were compared with the ones on the standard model. In the occlusal, mesial-medium and mesial-cervical segments, the double impression (DI) with condensation silicone presented similar values compared to the standard model. The values of the addition silicone with DI were similar to the standard model only in the mesial-occlusal segment. In the other segments (distal-cervical, distal-medium and distal-occlusal), all groups were statistically different from the control. It could be concluded that addition and condensation silicone impressions provided plaster dies with significant dimensional alterations in most of the evaluated areas when compared to the standard model. The retentive areas related to the onlay preparation influenced the dimensional stability of the addition and condensation silicone impressions.