Degradation products of resin-based materials detected in saliva in vivo.

OBJECTIVES: Dental composites remain under scrutiny regarding their (long-term) safety. In spite of numerous studies on the release of monomers both in vitro and in vivo, only limited quantitative data exist on the in vivo leaching of degradation products from monomers and additives. The aim of this observational study was for the first time to quantitatively and qualitatively monitor the release of parent compounds and their degradation products in saliva from patients undergoing multiple restorations. MATERIALS AND METHODS: Five patients in need of multiple large composite restorations (minimally 5 up to 28 restorations) due to wear (attrition, abrasion, and erosion) were included in the study, and they received adhesive restorative treatment according to the standard procedures in the university clinic for Restorative Dentistry. Saliva was collected at different time points, starting before the restoration up until 24 h after the treatment with composite restorations. Saliva extracts were analyzed by liquid chromatography-mass spectrometry. RESULTS: Leaching of monomers and degradation products was highest within 30 min after the placement of the restorations. The highest median concentrations of monomers were recorded for UDMA, BisEMA-3, and TEGDMA; yet, besides BisEMA-3 and TEGDMA, no monomers could be detected after 24 h. Mono- and demethacrylated degradation products remained present up to 24 h and concentrations were generally higher than those of monomers. In patients with multiple restorations, degradation products were still present in the sample taken before the next operation, several weeks after the previous operation. CONCLUSIONS: Exposure to residual monomers and degradation products occurs in the first hours after restoration. Monomers are present in saliva shortly after restoration, but degradation products can be detected weeks after the restoration confirming a long-term release. CLINICAL SIGNIFICANCE: Future research should focus more on the release of degradation products from monomers and additives from resin-based materials given their prolonged presence in saliva after restoration.

Agglomeration of titanium dioxide nanoparticles increases toxicological responses in vitro and in vivo.

BACKGROUND: The terms agglomerates and aggregates are frequently used in the regulatory definition(s) of nanomaterials (NMs) and hence attract attention in view of their potential influence on health effects. However, the influence of nanoparticle (NP) agglomeration and aggregation on toxicity is poorly understood although it is strongly believed that smaller the size of the NPs greater the toxicity. A toxicologically relevant definition of NMs is therefore not yet available, which affects not only the risk assessment process but also hinders the regulation of nano-products. In this study, we assessed the influence of NP agglomeration on their toxicity/biological responses in vitro and in vivo. RESULTS: We tested two TiO2 NPs with different primary sizes (17 and 117 nm) and prepared ad-hoc suspensions composed of small or large agglomerates with similar dispersion medium composition. For in vitro testing, human bronchial epithelial (HBE), colon epithelial (Caco2) and monocytic (THP-1) cell lines were exposed to these suspensions for 24 h and endpoints such as cytotoxicity, total glutathione, epithelial barrier integrity, inflammatory mediators and DNA damage were measured. Large agglomerates of 17 nm TiO2 induced stronger responses than small agglomerates for glutathione depletion, IL-8 and IL-1ß increase, and DNA damage in THP-1, while no effect of agglomeration was observed with 117 nm TiO2. In vivo, C57BL/6JRj mice were exposed via oropharyngeal aspiration or oral gavage to TiO2 suspensions and, after 3 days, biological parameters including cytotoxicity, inflammatory cell recruitment, DNA damage and biopersistence were measured. Mainly, we observed that large agglomerates of 117 nm TiO2 induced higher pulmonary responses in aspirated mice and blood DNA damage in gavaged mice compared to small agglomerates. CONCLUSION: Agglomeration of TiO2 NPs influences their toxicity/biological responses and, large agglomerates do not appear less active than small agglomerates. This study provides a deeper insight on the toxicological relevance of NP agglomerates and contributes to the establishment of a toxicologically relevant definition for NMs.

Filtration efficiency of surgical and FFP3 masks against composite dust.

The aim of this study was to investigate the protection efficiency of two types of face masks against composite dust and to characterize the particles that penetrated through the masks. Composite dust was created by grinding a commercial nano-filled composite in a plexiglass box without using water cooling or high vacuum evacuation, in order to obtain a worst-case exposure. Dust particles were collected using a personal inhalable aerosol sampler (IOM) fixed inside a custom-made phantom head. Surgical and filtering facepiece (FFP3) masks were tested, and the situation without a mask served as control. The IOM sampler contained a cassette with two filters to collect large inhalable (4-100 µm) and respirable dust particles (<4 µm). The amount of particles was determined gravimetrically by weighing filters before and after composite grinding, and further characterized by electron microscopy. Particle collection for both inhalable and respirable dust was the highest when no mask was used, and the lowest with the use of a FFP3 mask. Different sizes and shapes of particles were observed, with the largest particles (>1 µm) being seen when no mask was applied, whereas only nanoparticles could be detected when either type of face mask was applied. Even though FFP3 masks showed a higher filtration efficacy than surgical masks of the inhalable dust fraction, penetration of a small respirable particle fraction was inevitable for both masks.

Quick bonding using a universal adhesive.

OBJECTIVES: This study aimed to measure the microtensile bond strength (µTBS) of a new universal adhesive (UA) bonded to dentin following a quick bonding mode. MATERIALS AND METHODS: The experimental UA 'SKB-100,' now commercialized as Clearfil Universal Bond Quick ('C-UBq'; Kuraray Noritake), was investigated, with Scotchbond Universal ('SBU'; 3M Oral Care) and the two-step self-etch (SE) adhesive Clearfil SE Bond 2 ('C-SE2'; Kuraray Noritake) serving as references. The adhesives were employed separately in etch&rinse (E&R) and SE modes on each tooth half following a split-tooth design and following their respective instructions for use, except for C-UBq that besides the manufacturer's instructed quick bonding mode was also applied and left untouched for 20 s prior to light curing ('C-UBq_20s') and for C-SE2 that was also applied in the E&R mode. µTBS of half of the specimens was measured upon 1 week (1w), with the other half tested after 6-month (6m) storage in distilled water at 37 °C. Data were statistically analyzed using linear mixed-effects modeling and three-way ANOVA (p < 0.05). The adhesive-dentin interfaces were ultrastructurally characterized by TEM. RESULTS: C-UBq revealed a significantly lower µTBS than C-SE2. The highest µTBS was recorded for C-UBq_20s_1w and C-SE2_1w, both when applied in E&R mode. However, µTBS of C-UBq_20s significantly decreased upon aging in both modes. C-SE2 presented the significantly highest 'aged' µTBS. CONCLUSION: C-UBq applied in the quick bonding mode did not underperform to the UAs applied for 20 s. Superior bonding effectiveness was achieved by C-SE2 in SE and E&R bonding modes. CLINICAL RELEVANCE: Extended application time of C-UBq improved solely its immediate but not aged bonding effectiveness.

The effect of water spray on the release of composite nano-dust.

OBJECTIVE: To evaluate the collection efficiency of water spray on the release of airborne composite particles during grinding of composite materials. MATERIALS AND METHODS: Composite sticks (L:35 mm × W:5.4 mm × H:1.6 mm) of seven commercial dental composites were ground with a rough diamond bur (grain size 100 µm, speed 200,000 rpm). All experiments were performed in an enclosed 1-m3 chamber with low particulate background (< 1,000 #/cm3), and airborne particles were evaluated based on their electrical mobility. The number size distribution was determined by scanning mobility particle sizer (SMPS). Particles were collected by an electrostatic precipitator (ESP), and were ultramorphologically and chemically analyzed by a transmission electron microscope equipped with energy-dispersive X-ray spectroscopy (TEM-EDS). RESULTS: SMPS measurements confirmed that both dry and wet grinding generated high concentrations of nanoparticles particles with the highest concentration recorded during the last minute of grinding (1.80 × 106 - 3.29 × 106#/cm3), after which a gradual decline in particle concentration took place. Nevertheless, grinding with water spray resulted in a significant reduction of the number of released particles (5.6 × 105 - 1.37 × 106#/cm3). The smallest particle diameter was recorded during the last minute of grinding followed by a continuous growth for every next measurement. TEM of composite dust revealed a high concentration of particles varying in both size and shape. CONCLUSIONS: Regardless of whether the water cooling spray system was used during bur manipulation of composite materials, predominately nanoparticles were released. However, the particle concentrations were significantly decreased with water spray. CLINICAL RELEVANCE: Since water spray might not be sufficient in nanoparticle collection, special care should be taken to prevent inhalation of composite dust.

Secondary caries: prevalence, characteristics, and approach.

OBJECTIVES: The objectives of this cross-sectional survey were to determine the prevalence of secondary caries (SC) in general population, to identify patient- and material-related factors which may affect the prevalence, and to describe some clinical characteristics of SC lesions. MATERIALS AND METHODS: A total of 4036 restorations in 450 patients, who visited the university dental clinic for a regular (half) yearly checkup, were examined clinically (and radiographically) for the presence of SC. Clinical characteristics of the detected SC lesions (size, activity, and location) and the planned treatment were recorded. In addition, patients' caries-risk status was assessed according to the modified "cariogram" model. RESULTS: In total, 146 restorations were diagnosed with SC, which gives an overall prevalence of 3.6%. Restorative material, restoration class, patient's caries risk, and smoking habits were shown to be important factors, as SC prevalence was significantly higher with composites, class II restorations, high-caries-risk patients, and smokers. Restorations' gingival margins were most frequently affected by SC. The largest number of restorations with SC (72%) was scheduled for the replacement. CONCLUSIONS: Prevalence of SC was higher with composite than with amalgam restorations, irrespective of the patient's caries-risk status. Gingival margins of class II, including MOD restorations, seem to be the place of less resistance to SC development. Management of SC seems to place a considerable burden on the health care workforce and expenditure. CLINICAL RELEVANCE: Secondary caries (SC) is considered to be the main cause of dental restoration failure and one of the biggest clinical challenges related to dental composites. Nevertheless, its prevalence in daily practice is still not clear, which impedes an accurate estimation of its impact on health care costs.

Nine-year Clinical Performance of a HEMA-free One-step Self-etch Adhesive in Noncarious Cervical Lesions.

PURPOSE: One-step self-etch adhesives are among the most popular adhesives to directly bond composite to tooth tissue, mainly because of their perceived ease-of-use. The aim of this randomized controlled trial was to evaluate the clinical performance of a HEMA-free 1-step self-etch adhesive (1SEa), as compared with a 3-step etch-and-rinse adhesive (3E&Ra). MATERIALS AND METHODS: 267 non-carious cervical lesions in 52 patients were restored with a microhybrid composite Gradia Direct (GC), bonded in random order either with the HEMA-free 1SEa G-Bond (GB; GC) or the considered gold-standard 3E&Ra Optibond FL (OFL; Kerr). The restorations were evaluated after 6 months, 1, 2, 3, 5 and 9 years of clinical service for retention, marginal adaptation, marginal discoloration, caries occurrence and tooth sensitivity. Statistical analysis was done using a logistic regression model with generalized estimating equations (2-way GEE model). RESULTS: The patient recall rate at 9 years was 82.6%. The overall clinical success rate was 80.3% and 79.5% for GB and OFL, respectively. In total, 47 restorations (23 GB, 24 OFL) failed because of retention loss, severe marginal defects and/or discoloration and/or caries. A similar retention rate of 89.7% was recorded for both adhesives GB and OFL. Slight marginal defects and superficial marginal discoloration, though both still clinically acceptable, were significantly more observed at the incisal enamel side of GB (92.4% and 52.1%, respectively) than OFL (59.6%; 30.8%) restorations (p < 0.05). CONCLUSION: After 9 years of clinical service, class-V restorations bonded with the HEMA-free 1SEa performed clinically equally successful as those bonded with the 3E&Ra. However, more marginal deterioration, though still clinically acceptable, was recorded at the incisal enamel side for the 1SEa.

Epigenetic effects of carbon nanotubes in human monocytic cells.

Carbon nanotubes (CNTs) are fibrous carbon-based nanomaterials with a potential to cause carcinogenesis in humans. Alterations in DNA methylation on cytosine-phosphate-guanidine (CpG) sites are potential markers of exposure-induced carcinogenesis. This study examined cytotoxicity, genotoxicity and DNA methylation alterations on human monocytic cells (THP-1) after incubation with single-walled CNTs (SWCNTs) and multi-walled CNTs (MWCNTs). Higher cytotoxicity and genotoxicity were observed after incubation with SWCNTs than incubation with MWCNTs. At the selected concentrations (25 and 100 µg/ml), DNA methylation alterations were studied. Liquid chromatography-mass spectrometry (LC-MS/MS) was used to assess global DNA methylation, and Illumina 450K microarrays were used to assess methylation of single CpG sites. Next, we assessed gene promoter-specific methylation levels. We observed no global methylation or hydroxymethylation alterations, but on gene-specific level, distinct clustering of CNT-treated samples were noted. Collectively, CNTs induced gene promoter-specific altered methylation and those 1127 different genes were identified to be hypomethylated. Differentially methylated genes were involved in several signalling cascade pathways, vascular endothelial growth factor and platelet activation pathways. Moreover, possible contribution of the epigenetic alterations to monocyte differentiation and mixed M1/M2 macrophage polarisation were discussed.

Simultaneous analysis of bisphenol A based compounds and other monomers leaching from resin-based dental materials by UHPLC-MS/MS.

Resin-based dental materials have raised debates concerning their safety and biocompatibility, resulting in a growing necessity of profound knowledge on the quantity of released compounds into the oral cavity. In this context, the aim of this study was to develop a comprehensive and reliable procedure based on liquid chromatography with mass spectrometry for the simultaneous analysis of various leached compounds (including bisphenol A based compounds) in samples from in vitro experiments. Different experiments were performed to determine the optimal analytical parameters, comprising mass spectrometry parameters, chromatographic separation conditions, and sample preparation. Four internal standards were used as follows: deuterated diethyl phthalate and bisphenol A (commercially available), and deuterated analogues of triethylene glycol dimethacrylate and urethane dimethacrylate (custom-made). The optimized method was validated for linearity of the calibration curves and the associated correlation coefficient, lower limit of quantification, higher limit of quantification, and intra- and interassay accuracy and precision. Additionally, the developed liquid chromatography with tandem mass spectrometry method was applied to the analysis of leaching compounds from four resin-based dental materials. The results indicated that this method is suitable for the analysis of different target compounds leaching from dental materials. This method might serve as a valuable basis for quick and accurate quantification of leached compounds from resin-based dental materials in biological samples.

No evidence for the growth-stimulating effect of monomers on cariogenic Streptococci.

BACKGROUND: In spite of contradicting results, the high susceptibility of composites for secondary caries is still often associated with the bacterial growth-stimulating effect of released methacrylate monomers. However, most studies that showed this effect were performed with techniques having inherent limitations (spectrophotometry). OBJECTIVES: Therefore, our objective was to determine the effect of four methacrylate monomers (2-Hydroxyethyl methacrylate (HEMA), triethylene glycol dimethacrylate (TEGDMA), ethylene glycol dimethacrylate (EGDMA), diethylene glycol dimethacrylate (DEGDMA)) on the growth of two caries-associated bacteria, Streptococcus mutans and sobrinus, and one non-cariogenic species, Streptococcus sanguinis, using TaqMan quantitative polymerase chain reaction (qPCR) to quantify bacterial DNA. MATERIALS AND METHODS: Cultures were exposed to monomer solutions selected after spectrophotometric growth measurements. At baseline and predetermined time intervals, bacterial DNA was extracted and quantified with TaqMan qPCR. Biofilms grown in the presence of monomers were analyzed with scanning electron microscopy (SEM). RESULTS: Spectrophotometry indeed showed increased growth rates of all three strains with 5 mM TEGDMA, EGDMA, and DEGDMA and increased total biomass of S. sanguinis with 5 mM TEGDMA. However, qPCR failed to show any growth-stimulating effect of these monomers on S. mutans and S. sobrinus. In contrast, some monomers exhibited a growth-inhibiting effect on S. sanguinis. SEM revealed extracellular matter in S. sobrinus and S. sanguinis biofilms, which might be attributed to polymer formation. CONCLUSIONS: Techniques which quantify bacterial DNA are more appropriate to evaluate bacterial growth in the presence of monomers than spectrophotometry. CLINICAL RELEVANCE: Even though methacrylate monomers did not affect the growth of cariogenic species, growth inhibition of S. sanguinis, a non-cariogenic antagonistic species, may lead to ecological shifts towards higher cariogenicity.

Modified tricalcium silicate cement formulations with added zirconium oxide.

OBJECTIVES: This study aims to investigate the effect of modifying tricalcium silicate (TCS) cements on three key properties by adding ZrO2. MATERIALS AND METHODS: TCS powders were prepared by adding ZrO2 at six different concentrations. The powders were mixed with 1 M CaCl2 solution at a 3:1 weight ratio. Biodentine (contains 5 wt.% ZrO2) served as control. To evaluate the potential effect on mechanical properties, the mini-fracture toughness (mini-FT) was measured. Regarding bioactivity, Ca release was assessed using ICP-AES. The component distribution within the cement matrix was evaluated by Feg-SEM/EPMA. Cytotoxicity was assessed using an XTT assay. RESULTS: Adding ZrO2 to TCS did not alter the mini-FT (p = 0.52), which remained in range of that of Biodentine (p = 0.31). Ca release from TSC cements was slightly lower than that from Biodentine at 1 day (p > 0.05). After 1 week, Ca release from TCS 30 and TCS 50 increased to a level that was significantly higher than that from Biodentine (p < 0.05). After 1 month, Ca release all decreased (p < 0.05), yet TCS 0 and TCS 50 released comparable amounts of Ca as at 1 day (p > 0.05). EPMA revealed a more even distribution of ZrO2 within the TCS cements. Particles with an un-reacted core were surrounded by a hydration zone. The 24-, 48-, and 72-h extracts of TCS 50 were the least cytotoxic. CONCLUSIONS: ZrO2 can be added to TCS without affecting the mini-FT; Ca release was reduced initially, to reach a prolonged release thereafter; adding ZrO2 made TCS cements more biocompatible. CLINICAL RELEVANCE: TCS 50 is a promising cement formulation to serve as a biocompatible hydraulic calcium silicate cement.

Effect of Bulk-filling on the Bonding Efficacy in Occlusal Class I Cavities.

PURPOSE: To evaluate the effect of bulk filling Class I posterior cavities on bonding to cavity-bottom dentin. MATERIALS AND METHODS: Two flowable "base" bulk-fill composites (Filtek Bulk Fill Flowable, SDR), one paste-like "full-body" bulk-fill composite (Tetric EvoCeram Bulk Fill) and one conventional paste-like composite (Filtek Z100) were bonded (G-ænial Bond) to either a flat surface (3.5 x 3.5 x 4 mm; C-factor: 0.18) or a Class I cavity (3.5 x 3.5 x 4 mm; C-factor: 5.8). After 1-week water storage, the restorations were sectioned to obtain 4 rectangular microspecimens that were subjected to microtensile bond strength (µTBS) testing. RESULTS: No significant differences in µTBS were recorded between all composites when bonded onto a flat surface (p > 0.05). When bonded into a Class I cavity, the µTBS of all composites except SDR significantly decreased (p < 0.001). CONCLUSION: Both the configuration factor and the type of bulk-fill composite were found to have a great impact on bonding to cavity-bottom dentin.

Correlative micro-Raman/EPMA analysis of the hydraulic calcium silicate cement interface with dentin.

OBJECTIVES: This study aims to characterize the chemical interplay of hydraulic calcium silicate cements at dentin. MATERIALS AND METHODS: Class I cavities were prepared in non-carious human third molars and filled with Biodentine (Septodont) or ProRoot MTA (Dentsply). After 1-day, 1-week, and 1-month Dulbecco's phosphate-buffered saline (DPBS) storage, the specimens were cross-sectioned perpendicular to the cement-dentin interface. The interfaces were evaluated using micro-Raman (µRaman) spectroscopy and at a higher spatial resolution using field emission gun electron probe microanalysis (Feg-SEM/EPMA). RESULTS: µRaman spectroscopy revealed the formation of a transition zone at the interface of both Biodentine (Septodont) and ProRoot MTA (Dentsply) with dentin, having an average thickness of, respectively, 7.5 ± 4.2 and 6.2 ± 5.4 µm, which however was not statistically different. No difference in interfacial ultrastructure and chemistry was found using µRaman spectroscopy between 1 day, 1 week, and 1 month DPBS-stored specimens. The observation of a transition zone at the cement-dentin interfaces contrasts with the EPMA data that revealed a sharper transition from cement to dentin. Again, no difference in interfacial ultrastructure and chemistry was found for different storage periods, with the exception of one 1 month DPBS-stored specimen prepared using Biodentine (Septodont). More specifically, EPMA revealed a gap of about 10-µm wide in the latter specimen that was filled up with newly formed calcium phosphate depositions. CONCLUSIONS: Up to 1 month, the interaction of hydraulic calcium silicate cements investigated did not reveal ultrastructural or chemical changes at unaffected dentin with the exception of a calcium phosphate gap-filling property. CLINICAL RELEVANCE: Hydraulic calcium silicate cements were found to fill gaps by calcium phosphate deposition, however, without conducting chemical changes to the adjacent dentin.

Five-year clinical performance of a HEMA-free one-step self-etch adhesive in noncarious cervical lesions.

OBJECTIVES: The objective of this randomized controlled trial was to evaluate the clinical performance of a 2-hydroxyethyl methacrylate (HEMA)-free one-step adhesive. MATERIALS AND METHODS: Two hundred sixty-seven cervical lesions in 52 patients were restored with the composite Gradia Direct (GC), bonded with either the one-step self-etch adhesive G-Bond (GC) or the three-step etch-and-rinse adhesive Optibond FL (Kerr) in a random order. The restorations were evaluated for retention, marginal integrity, marginal discoloration, and caries occurrence after 5 years. Specific statistics were used to account for the clustered data (multiple restorations per patient). RESULTS: The clinical success rate for G-Bond (87.4 %) was not significantly different from that of Optibond FL (90.9 %). Both adhesives showed progressive marginal deterioration, but G-Bond exhibited more small enamel defects and marginal discoloration, and unlike previous recalls, several restorations failed because of deep microleakage. Large sclerotic lesions were a significant risk factor for retention loss with G-Bond. Irrespective of the adhesive, almost all restorations with retention loss were located in the lower jaw. CONCLUSION: After 5 years of clinical service, restorations bonded with the HEMA-free one-step adhesive did not need repair or replacement more often than those with the three-step etch-and-rinse adhesive, and both adhesives had a high retention rate (>90 %). There were indications that G-Bond did not (self-)etch enough in some clinical situations, as G-Bond exhibited more incisal defects and marginal discolorations, and sclerotic lesions were at higher risk of retention loss. CLINICAL RELEVANCE: The clinical performance of the HEMA-free one-step adhesive was clinically acceptable after 5 years.

Elution from direct composites for provisional restorations.

Purpose To assess elution from direct composite materials for provisional restorations and compare them with elution from direct restorative composites for permanent restorations.Methods Two dual-cure (Integrity Multi-Cure and Tempsmart DC) and two self-curing composites (Protemp 4 and Structur 3) were used, with Essentia serving as a reference. Cylindrical specimens (n=20) were cured according to the manufacturer's instructions; the dual-cure materials were prepared in both self- and dual-curing modes. Elution experiments were performed using water and absolute ethanol. The samples were incubated at 37 °C for either 24 h or four weeks; the extraction solvents were refreshed weekly. The eluted BisEMA (-3 / -6 / -10), BisGMA, CQ, UDMA, and TEGDMA were quantified using UHPLC-MS/MS.Results Monomer elution was detected in all provisional composites at 24 h and four weeks, but the amounts released did not exceed those released by the reference composite. When prepared in self-curing mode, Integrity Multi-Cure exhibited significantly higher elution of BisEMA-3, -6, and -10 in ethanol both after 24 h and cumulatively after four weeks. Self-cured Tempsmart DC released significantly more CQ, TEGDMA, and UDMA in both water and ethanol after immersion for 24 h and four weeks, along with significantly more BisGMA in ethanol both after 24 h and four weeks comparison to dual-cured Tempsmart DC (two-way ANOVA, post-hoc Tukey, P < 0.05).Conclusions Provisional composite materials did not elute higher amounts of monomers than a restorative composite. Dual-cured materials, prepared in the self-curing mode, show a trend towards higher monomer elution.

Level of BPA contamination in resin composites determines BPA release.

OBJECTIVES: Resin composites may release bisphenol A (BPA) due to impurities present in the monomers. However, there is a lack of knowledge regarding the leaching characteristics of BPA from resin composites. Therefore, experimental resin composites were prepared with known amounts of BPA. The objective of this study was (1) to determine which amount of BPA initially present in the material leaches out in the short term and, (2) how this release is influenced by the resin composition. METHODS: BPA (0, 0.001, 0.01, or 0.1 wt%) was added to experimental resin composites containing 60 mol% BisGMA, BisEMA(3), or UDMA, respectively, as base monomer and 40 mol% TEGDMA as diluent monomer. Polymerized samples (n = 5) were immersed at 37 °C for 7 days in 1 mL of water, which was collected and refreshed daily. BPA release was quantified with UPLC-MS/MS after derivatization with pyridine-3-sulfonyl chloride. RESULTS: Between 0.47 to 0.67 mol% of the originally added BPA eluted from the resin composites after 7 days. Similar elution trends were observed irrespective of the base monomer. Two-way ANOVA showed a significant effect of the base monomer on BPA release, but the differences were small and not consistent. SIGNIFICANCE: The released amount of BPA was directly proportional to the quantity of BPA present in the resin composite as an impurity. BPA release was mainly diffusion-based, while polymer composition seemed to play a minor role. Our results underscore the importance for manufacturers only to use monomers of the highest purity in dental resin composites to avoid unnecessary BPA exposure in patients.

Hydrolytic stability of three-step etch-and-rinse adhesives in occlusal class-I cavities.

OBJECTIVES: A dental adhesive without small and hydrophilic monomers such as 2-hydroxyethyl methacrylate (HEMA) and triethylene glycol dimethacrylate (TEGDMA) would be beneficial in order to avoid contact allergies. However, these monomers are important to increase infiltration and polymerization of the adhesive. Therefore, the purpose of this study was to evaluate the bonding effectiveness and bond durability of a more hydrophobic and biocompatible adhesive as compared to a conventional three-step etch-and-rinse adhesive. METHODS: Sixteen non-carious human third molars were used to determine the micro-tensile bond strength testing (µTBS) and interfacial ultrastructure by transmission electron microscopy (TEM) of the more hydrophobic cmf adhesive system (Saremco) adhesive as compared to the control OptiBond FL (Kerr). RESULTS: The more hydrophobic and biocompatible three-step etch-and-rinse adhesive was able to produce a reasonable short-time bonding effectiveness. In the long term, the collagen fibrils in the hybrid layer were not effectively protected and were prone to hydrolytic degradation. As a result, long-term bonding effectiveness of this novel adhesive was very low. CONCLUSIONS: Application of a more hydrophobic adhesive without altering the application procedure considerably results in a reduced durability of the created bond CLINICAL RELEVANCE: Omitting small and hydrophilic components from the adhesive formulation may impair the durability of your composite restoration.

Fourteen-year Clinical Performance of a HEMA-free One-step Self-etch Adhesive in Non-carious Cervical Lesions.

PURPOSE: This randomized controlled trial aimed to evaluate the 14-year clinical performance of a HEMA-free 1-step self-etch adhesive (1SEa) compared with that of a 3-step etch-and-rinse adhesive (3E&Ra). MATERIALS AND METHODS: 267 non-carious cervical lesions in 52 patients were restored with the microhybrid composite Gradia Direct (GC), bonded in random order either with the HEMA-free 1SEa G-Bond (GB; GC) or the 3E&Ra Optibond FL (OFL; Kerr), which is considered the gold-standard E&Ra (control). The restorations were followed over 14 years for retention, marginal adaptation and discoloration, and caries occurrence. Statistical analysis involved a logistic regression model with generalized estimating equations (2-way GEE model). RESULTS: The patient recall rate at 14 years was 63%. In total, 79 restorations (39 GB, 40 OFL) failed because of retention loss (GB: 19.4%, OFL: 19.6%), severe marginal defects, discoloration and/or caries (GB: 21.7%; OFL: 22.5%). The overall clinical success rate was 58.9% and 57.9% for GB and OFL, respectively. The number of restorations with an unacceptable marginal defect (GB: 14.5%; OFL: 19.2%) and deep marginal discoloration (GB: 18.2%; OFL: 13.2%) increased during the last 5 years. No significant difference in overall clinical performance was recorded between the two adhesives (p > 0.05). Changes in the medical health of some patients and recurrence of abrasion/erosion/abfraction increased the failure rate and retention rate. CONCLUSION: After 14 years, restorations bonded with the HEMA-free 1SEa performed as well as those bonded with the 3E&Ra gold standard. Unacceptable marginal deterioration was the main reason for failure, followed by loss of retention.

Contamination of nanoparticles by endotoxin: evaluation of different test methods.

BACKGROUND: Nanomaterials can be contaminated with endotoxin (lipopolysaccharides, LPS) during production or handling. In this study, we searched for a convenient in vitro method to evaluate endotoxin contamination in nanoparticle samples. We assessed the reliability of the commonly used limulus amebocyte lysate (LAL) assay and an alternative method based on toll-like receptor (TLR) 4 reporter cells when applied with particles (TiO(2), Ag, CaCO(3) and SiO(2)), or after extraction of the endotoxin as described in the ISO norm 29701. RESULTS: Our results indicate that the gel clot LAL assay is easily disturbed in the presence of nanoparticles; and that the endotoxin extraction protocol is not suitable at high particle concentrations. The chromogenic-based LAL endotoxin detection systems (chromogenic LAL assay and Endosafe-PTS), and the TLR4 reporter cells were not significantly perturbed. CONCLUSION: We demonstrated that nanoparticles can interfere with endotoxin detection systems indicating that a convenient test method must be chosen before assessing endotoxin contamination in nanoparticle samples.

Nano-titanium dioxide modulates the dermal sensitization potency of DNCB.

We determined the ability of a model nanoparticle (NP) (titanium dioxide, TiO(2)) to modulate sensitization induced by a known potent dermal sensitizer (dinitrochlorobenzene) using a variant of the local lymph node assay called lymph node proliferation assay.BALB/c mice received sub-cutaneous injections of vehicle (2.5 mM sodium citrate), TiO(2) NPs (0.004, 0.04 or 0.4 mg/ml) or pigment particles (0.04 mg/ml) both stabilized in sodium citrate buffer at the base of each ear (2x50µl), before receiving dermal applications (on both ears) of 2,4-Dinitrochlorobenzene (DNCB) (2x25µl of 0.1%) or its vehicle (acetone olive oil - AOO (4:1)) on days 0, 1 and 2. On day 5, the stimulation index (SI) was calculated as a ratio of (3)HTdR incorporation in lymphocytes from DNBC-treated mice and AOO-treated controls. In a second experiment the EC(3)-value for DNCB (0 to 0.1%) was assessed in the absence or presence of 0.04 mg/ml TiO(2). In a third experiment, the lymphocyte subpopulations and the cytokine secretion profile were analyzed after TiO(2) (0.04 mg/ml) and DNCB (0.1%) treatment. Injection of NPs in AOO-treated control mice did not have any effect on lymph node (LN) proliferation. DNCB sensitization resulted in LN proliferation, which was further increased by injection of TiO(2) NPs before DNCB sensitization. The EC(3) of DNCB, with prior injection of vehicle control was 0.041%, while injection with TiO(2) decreased the EC(3) of DNCB to 0.015%. TiO(2) NPs pre-treatment did not alter the lymphocyte subpopulations, but significantly increased the level of IL-4 and decreased IL-10 production in DNCB treated animals.In conclusion, our study demonstrates that administration of nano-TiO(2) increases the dermal sensitization potency of DNCB, by augmenting a Th(2) response, showing the immunomodulatory abilities of NPs.