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.

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.

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.

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.

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.

A 13-year clinical evaluation of two three-step etch-and-rinse adhesives in non-carious class-V lesions.

This 13-year randomized clinical trial compared the clinical effectiveness of two three-step etch-and-rinse adhesives in combination with a hybrid, stiffer composite versus a micro-filled, more flexible composite. The influence of composite stiffness on the clinical performance of one of the adhesives was assessed as well. One hundred and forty-two non-carious cervical lesions were restored with composites with contrasting stiffness. Seventy-one patients randomly received two cervical restorations placed following two out of three adhesive procedures: (1) the three-step etch-and-rinse adhesive Permaquick applied with the stiff micro-hybrid composite Amelogen Hybrid (PMQ-H, Ultradent), (2) Permaquick applied with the more flexible micro-filled Amelogen Microfill (PMQ-M, Ultradent), or (3) the "gold-standard" three-step etch-and-rinse adhesive Optibond FL applied with the micro-hybrid composite Prodigy (OFL-P, Kerr). The restorations were evaluated after 6 months, 1, 2, 3, 5, 7, and 13 years of clinical service regarding their retention, marginal integrity and discoloration, caries occurrence, preservation of tooth vitality, and post-operative sensitivity. Retention loss, severe marginal defects, and/or discoloration that needed intervention (repair or replacement) and the occurrence of caries were considered as clinical failures. The recall rate at 13 years was 77%. Bond degradation after 13 years was mainly characterized by a further increase in the presence of small but clinically acceptable marginal defects and superficial marginal discoloration. Twelve percent of the OFL-P restorations were clinically unacceptable. In the PMQ group, 22% of the PMQ-M restorations and 26% of the PMQ-H restorations needed repair or replacement. Regarding the clinical failure rate, Optibond FL scored significantly better than Permaquick (McNemar; p = 0.015). No statistically significant differences were found between the micro-filled and the hybrid composite for each of the parameters evaluated (McNemar, p > 0.05). After 13 years of clinical functioning, the clinical effectiveness of the three adhesive/composite combinations remained highly acceptable.

Clinical effectiveness of a one-step self-etch adhesive in non-carious cervical lesions at 2 years.

A 2-year randomized, controlled prospective study evaluated the clinical effectiveness of a one-step self-etch adhesive and a "gold-standard" three-step etch-and-rinse adhesive in non-carious Class-V lesions. The null hypothesis tested was that the one-step self-etch adhesive does perform clinically equally well as the three-step etch-and-rinse adhesive. A total of 161 lesions in 26 patients were restored with Clearfil AP-X (Kuraray). The restorations were bonded either with the "all-in-one" adhesive Clearfil S3 Bond (Kuraray) or with the three-step etch-and-rinse adhesive Optibond FL (Kerr). The restorations were evaluated at baseline and after 6 months, 1 and 2 years, regarding their retention, marginal adapation, marginal discoloration, caries occurrence, preservation of tooth vitality and post-operative sensivity. Retention loss, severe marginal defects and/or discoloration that needed intervention (repair or replacement) and the occurrence of caries were considered as clinical failures. The recall rate at 2 years was 93.8%. Only one Clearfil S3 Bond restoration was lost at the 2-year recall. All other restorations were clinically acceptable. The number of restorations with defect-free margins decreased severely during the 2-year study period (to 6.7% and 25.3% for Clearfil S3 Bond and Optibond FL, respectively). The Clearfil S3 Bond restorations presented significantly more small marginal defects at the enamel side than the Optibond FL restorations (Clearfil S3 Bond: 93.3%; Optibond FL: 73.3%; p = 0.000). Superficial marginal discoloration increased in both groups (to 53.3% and 36% for Clearfil S3 Bond and Optibond FL, respectively) and was also more pronounced in the Clearfil S3 Bond group (p = 0.007). After 2 years, the simplified one-step self-etch adhesive Clearfil S3 Bond and the three-step etch-and-rinse adhesive Optibond FL were clinically equally successful, even though both adhesives were characterized by progressive degradation in marginal integrity. Clearfil S3 Bond exhibited more small enamel marginal defects and superficial marginal discolorations.

Identification of chemicals leaching from dental resin-based materials after in vitro chemical and salivary degradation.

OBJECTIVES: Only little is known about degradation of methacrylate monomers. Therefore, using in vitro chemical and saliva degradation this study aimed to identify the degradation products of organic compounds present in resin-based dental materials. METHODS: Ten dental monomers and nine polymerized dental resin-based materials were immersed for 24 h in chemical media (0.1 M HCl, 0.1 M NaOH) and human pooled saliva in order to identify leached monomers and degradation products from chemical and saliva degradation. Samples were analyzed using liquid chromatography coupled to high-resolution mass spectrometry to identify previously unknown degradation products. RESULTS: During in vitro chemical degradation, uncured monomers were rapidly hydrolyzed into mono- and demethacrylated degradation products. During chemical degradation in alkaline conditions of polymerized materials, considered the worst-case scenario, only degradation products could be detected. In acidic conditions, monomers and their degradation products were detected. In addition, different additives such as EDMAB, DMPA and HMBP were present in acidic degradation samples. Degradation in human pooled saliva for 24 h to mimic the in vivo situation, resulted in the identification of both monomers and their degradation products. CLINICAL SIGNIFICANCE: Using state-of-the-art high-resolution mass spectrometry previously unknown degradation products of commonly used monomers were identified for the first time. Results show that patients may be exposed to monomers and their degradation products in the first 24 h after restorative procedures. The results provide a base for further research on the degradation of resin-based dental composites in order to assess their safety using elution and toxicity studies.

Bisphenol A release from short-term degraded resin-based dental materials.

OBJECTIVES: There is still much debate about the release of bisphenol A (BPA) from resin-based dental materials. Therefore, this study aimed to quantify BPA present as an impurity and to evaluate whether their degradation by salivary, bacterial, and chemical challenges could increase its release. METHODS: BPA was determined in three different amounts (300, 400, and 500 µg) of eight unpolymerized resin-based materials (four composites, one fissure sealant, two adhesives and one root canal sealer). Next, polymerized samples (n = 5) of each material were immersed in 1 mL of whole human pooled saliva collected from adults, Streptococcus mutans (2 × 107 CFU/mL), and acidic (0.1 M HCl), alkaline (0.1 M NaOH), and control media, respectively. The amount of BPA was quantified using an UPLC-MS/MS method including derivatization of BPA by pyridine-3-sulfonyl chloride. RESULTS: Only the composites contained trace amounts of BPA above the limit of quantification (ranging from 301±32 pg PBA/mg to 1534±62 pg BPA/mg), most likely as impurity from the synthesis of the monomers. The amounts of BPA released from polymerized materials upon salivary and bacterial degradation were too low for accurate quantification, but in water, quantifiable amounts of BPA were released from all materials. In alkaline media, the BPA release from two composites was significantly decreased, while the release from one adhesive was significantly increased, compared to water. CONCLUSIONS: BPA already present in unpolymerized resin-based materials may account for the release of BPA after polymerization. There was no clear indication that short-term material degradation leads to increased release of BPA.

Three-year clinical performance of a HEMA-free one-step self-etch adhesive in non-carious cervical lesions.

Despite the fact that one-step adhesives are currently used routinely in clinical practice, long-term studies on their clinical performance are scarce. The objective of this randomized controlled clinical trial was to test the hypothesis that a 2-hydroxyethyl methacrylate (HEMA)-free one-step self-etch adhesive performs worse than a conventional multistep etch-and-rinse adhesive. Two-hundred and seventy-six non-carious cervical lesions in 52 patients were restored with a micro-hybrid composite (Gradia Direct; GC). These restorations were bonded in random order either with the HEMA-free one-step adhesive G-Bond (GC) or with the 'gold-standard' (control) three-step adhesive Optibond FL (Kerr). The restorations were evaluated after 6, 12, 24, and 36 months of clinical service regarding retention, marginal adaptation, microleakage, caries occurrence, and sensitivity. After a medium-long period of 3 yr, similar success in clinical performance was observed for the simplified all-in-one adhesive and the conventional three-step adhesive. However, the one-step adhesive exhibited significantly more incisal marginal defects and discolorations. Whereas marginal degradation appeared to arrest for the multistep etch-and-rinse adhesive after 12 months, the enamel margins of the restorations bonded with the one-step self-etch adhesive continued to deteriorate. These incisal marginal defects were, however, small and could easily be removed by polishing. For both adhesives, large and sclerosed lesions appeared to be at higher risk of retention loss.

Optimizing glass-ceramic bonding incorporating new silane technology in an experimental universal adhesive formulation.

OBJECTIVE: Incorporating silane-coupling agent into universal adhesives (UAs) to simplify adhesive luting of glass-ceramic restorations appeared ineffective due to silane's instability in an acidic aqueous solution. This study aimed to evaluate new silane technology added to an experimental UA to be bonded to glass ceramics without separate prior silanization. METHODS: Combined silane technology, consisting of 3-(aminopropyl)triethoxysilane (APTES) and γ-methacryloxypropyltriethoxysilane (γMPTES), was incorporated into an experimental UA formulation, being referred to as ADH-XTE (3M Oral Care). Immediate and aged shear bond strength (SBS) of ADH-XTE onto as-milled ('AM'), tribochemical silica-coated ('TSC'), HF-etched ('HF'), and mirror-polished ('MP') glass-ceramic CAD/CAM blocks (IPS e.max CAD) with/without separate silanization was measured (n = 10/group). The control adhesives included Scotchbond Universal ('SBU') and Scotchbond 1 XT ('SB1-XT'). The glass-ceramic surface topography and the fractography of the SBS-debonded specimens were observed by SEM. RESULTS: Without separate prior silanization, the experimental UA ADH-XTE, containing combined APTES/γMPTES silane technology, significantly outperformed the glass-ceramic bonding efficiency of its silane-containing SBU precursor, while it performed equally effective as SBU applied with prior silanization. Upon aging, significant reduction in SBS was recorded when ADH-XTE was bonded to TSC glass-ceramic surfaces (p < 0.05), while not to HF ones. Notably, the lowest SBS was obtained when the UAs were bonded to AM and MP glass-ceramic surfaces, in particular when applied without separate prior silanization (p < 0.05). SIGNIFICANCE: The glass-ceramic bonding capacity of the new combined APTES/γMPTES silane-containing UA ADH-XTE surpassed that of its SBU precursor. HF etching remains needed to durably bond to glass-ceramics.

Assessing the estrogenic activity of chemicals present in resin based dental composites and in leachates of commercially available composites using the ERα-CALUX bioassay.

OBJECTIVE: The biocompatibility of resin based dental composites has not yet been fully characterized even though certain monomers used in these composites are synthesized from Bisphenol A (BPA), a well-known estrogenic endocrine disruptor. As a result, they show structural relationship to BPA and can contain it as an impurity. Therefore, the estrogenic activity of 9 monomers, 2 photoinitiators, one photostabilizer and leachates of 4 commercially available composites was determined. METHODS: The ERα-CALUX bioassay was used to determine both agonistic and antagonistic estrogenic activities of the pure compounds (BPA, BisDMA, BisGMA, BisEMA(3), BisEMA(6), BisEMA(10), TEGDMA, TCD-DI-HEA, BADGE, UDMA, HMBP, DMPA, CQ) and the leachates of cured composite disks. The leachates of 4 commercially available composites (Solitaire 2, Ceram.x Spectra ST, G-ænial Posterior and Filtek Supreme XTE) in water and 0.1 M NaOH (pH = 13, 'worst-case scenario') were tested for estrogenic activity (pooled leachates from 10 cured composite disks). RESULTS: Agonistic estrogenic activity was found for the monomer BisDMA, the photostabilizer HMBP and photoinitiator DMPA. All leachates from the 4 tested composites showed significant agonistic estrogenic activity higher than the DMSO control, and the highest activity (potency and efficacy) was found for Solitaire 2, followed by Ceram.x Spectra ST. Furthermore, antagonistic estrogenic activity was found in the leachates from G-ænial Posterior. SIGNIFICANCE: These results show that significant estrogenic activity was found in all leachates of the cured composite disks, and that this estrogenicity is most likely due to a mixture effect of multiple estrogenic compounds (including BPA, HMBP and DMPA). This indicates that further research into the endocrine activity of all the compounds that are present in these composites (even at low quantities) and their possible mixture effect is warranted to guarantee their safe use.