Leaching components and initial biocompatibility of novel bioactive restorative materials.

OBJECTIVES: Bioactive restorative materials were developed on the premise that direct restorations should not only serve the purpose of reconstructing dental hard tissue defects but also exhibit biological features that prevent secondary caries development, without having adverse effects on the host cells. This study focuses on assessing the in vitro biocompatibility of two novel bioactive restorative materials. METHODS: Specimens of the bioactive restorative materials, Cention Forte (CF) and ACTIVA BioACTIVE RESTORATIVE (AB), a glass ionomer cement/glass hybrid (EQUIA Forte HT, EF) and an established nanohybrid composite (Venus Diamond, VD) were produced and finished. The specimens were eluted in water and methanol and the resulting eluates were analyzed via gas chromatography-mass spectrometry. hGF-1 cells were exposed to eluates prepared in cell culture medium. Cellular ATP levels, oxidized glutathione concentration, caspase-3/7 activity and the inflammatory response (IL-6 and PGE2 levels) were determined. Microscopic images were taken to examine the cell morphology. RESULTS: Methyl methacrylate and 2-Hydroxyethyl methacrylate were the main monomers detected in CF and AB eluates. All materials inhibited cell proliferation and led to significantly reduced ATP-levels. The cells exhibited a healthy morphology in the presence of CF and AB. Cells exposed to VD showed increased oxidized glutathione levels. Only EF led to enhanced caspase-3/7 activity. CF and AB caused IL-6 levels to increase, while EF and AB led to enhanced PGE2 levels. SIGNIFICANCE: CF and AB are promising materials from a biological point of view and seem to have improved bioactive properties compared to glass ionomer cements.

Disrupted biomineralization in zebra mussels after exposure to bisphenol-A: Potential implications for molar-incisor hypomineralization.

OBJECTIVE: The aetiology of molar-incisor hypomineralization (MIH) is currently unclear. A major hurdle in MIH research is the lack of adequate model systems. The study investigated the feasibility of zebra mussel (Dreissena polymorpha) as a novel model to screen potential MIH-related factors. METHODS: In four experiments with overall 46 groups (n = 7 mussels/group), six groups per experiment were incubated with 100 mg/l calcein (mineralization marker) solution for 96 h to evaluate the dynamics of shell biomineralization, another six groups with tap water only (controls). Then zebra mussels with and without calcein pre-incubation were exposed to cadmium sulfate hydrate (3CdSO4•8H2O) (positive control; 0, 0.01, 0.1, 1, 10 and 100 mg/l), possible aetiological factors of MIH including bisphenol-A (BPA; 0, 0.02, 0.2, 2, 20 and 200 mg/l) and erythromycin (0, 0.1, 1, 10, 100 and 1000 mg/l) as mineralization "disruptors", and doxycycline (0, 0.1, 1, 10, 100 and 1000 mg/l) for 96 h, respectively. After two weeks, the mussels were sacrificed and shells were embedded in methylmethacrylate for fluorescence intensity analysis. RESULTS: Mortality rate was 100% after 20, 200 mg/l BPA and 10, 100 mg/l 3CdSO4•8H2O exposure. Thereby, the median lethal concentration (96 h-LC50) of BPA was 6.3 mg/l (95% CI, 1.3-34.4 mg/l), and that of cadmium was 3.1 mg/l (95% CI, 0.7-10.5 mg/l). Notably, calcein fluorescence in shells significantly decreased (p < 0.05) after 2 mg/l BPA and 1 mg/l 3CdSO4•8H2O exposure. SIGNIFICANCE: These findings suggest that BPA may disrupt biomineralization. Biomineralization in zebra mussels seems to be an effective model for investigating potential MIH-related factors.

Intranuclear cell uptake and toxicity of titanium dioxide and zirconia particles as well as bacterial adhesion on dental titanium- and zirconia-implants.

OBJECTIVE: Previous studies have shown that particles can be released from dental titanium (Ti)- and zirconia (ZrO2)-implants. Titanium dioxide (TiO2)- and ZrO2-particles were compared regarding their toxicity and intranuclear cell uptake as well as the adhesion of various anaerobic bacteria on Ti- and ZrO2-implants. METHODS: Cyto- and genotoxicity of TiO2-microparticles (TiO2-MPs) and TiO2-nanoparticles (TiO2-NPs) in periodontal ligament (PDL)-hTERT cells were determined with XTT test and DNA damage with comet assay. Particle sizes of TiO2- and ZrO2-particles were measured with scanning electron microscope. Intranuclear uptake in PDL-hTERT cells was determined with laser scanning confocal microscopy. Adhesions of relevant anaerobic mouth bacteria Porphyromonas gingivalis, Prevotella intermedia and Aggregatibacter actinomycetemcomitans on Ti- and ZrO2-implants were investigated by cultivation and counting bacterial colonies. RESULTS: Particle size measurements revealed that 99% of the TiO2-NPs had a size below 100 nm and 88% of the TiO2-MPs sizes were between 50 and 200 nm. Following EC50 values were found for particles (mg/l): 92 (TiO2-MPs) and 15 (TiO2-NPs). A significant increase in olive tail moment (OTM) was found for TiO2-NPs at a concentration of 1/10 EC50. TiO2- and ZrO2-NPs had a higher intranuclear cell uptake efficiency, compared to corresponding TiO2- and ZrO2-MPs. All investigated particles could be detected in cell nucleus. Adhesion of all investigated bacterial species was significantly higher on Ti-implants, compared to ZrO2-implants. CONCLUSION: Ti usually develops an oxide layer (TiO2). Particles released from Ti-implants should be TiO2-particles or Ti-particles coated with a TiO2-layer. Toxicity of released Ti-particles depends on their oxidation state and on their size (NP or MP). Particularly, NPs were more cyto- and genotoxic compared to the corresponding MPs. TiO2- and ZrO2-NPs showed a significant increase in the intranuclear cell uptake ratio at higher exposure concentration, compared to lower concentrations and consequently might lead to a higher potential of DNA damage. Adhesion of bacteria to ZrO2-implants is reduced, compared to Ti-implants. Therefore, ZrO2-implants might contribute to reduced biological complications (e.g. periimplantitis).

Salivary and gingival CXCL8 correlation with periodontal status, periodontal pathogens, and smoking.

OBJECTIVES: Neutrophil granulocytes have been proposed to play a major role in the mediation of periodontitis-associated tissue destruction. Their recruitment and activation are regulated by the chemokine CXCL8. This study aimed to delineate the dependency of CXCL8 expression in gingival crevicular fluid (GCF) and saliva on periodontal status, bacterial infection, and smoking, in patients with periodontitis. METHODS: The study cohort comprised 279 subjects with untreated periodontitis. Probing pocket depth (PPD), gingival recession, bleeding on probing (BOP), plaque index, and bone loss were evaluated. CXCL8 was determined in saliva and GCF using flow cytometry. RESULTS: Considering the entire study sample, CXCL8 levels were correlated with the mean PPD (ρ = 0.131; p = 0.029), severity of periodontitis (ρ = 0.121; p = 0.043), BOP (ρ = 0.204; p = 0.001), and smoking (ρ = -0.219; p < 0.0001) in GCF; and, in whole saliva, with mean PPD (ρ = 0.154; p = 0.010) severity of periodontitis (ρ = 0.140; p = 0.020), gender (ρ = 0.178; p = 0.003), and smoking (ρ = -0.156; p = 0.010). Subgroup analysis among non-smokers revealed significantly higher amounts of CXCL8 in GCF (p = 0.012) and saliva (p = 0.026) comparing subjects with mean PPD ≤3mm and >3mm. CONCLUSION: The current study revealed a strong dependency of CXCL8 expression in GCF on the severity and activity of periodontal disease. Smoking causes a significant reduction in CXCL8 expression in saliva and GCF.

Elution behavior of a 3D-printed, milled and conventional resin-based occlusal splint material.

OBJECTIVE: The elution of unpolymerized (co-)monomers and additives from methacrylic resin-based materials like polymethyl methacrylate (PMMA) can cause adverse side effects, such as mutagenicity, teratogenicity, genotoxicity, cytotoxicity and estrogenic activity. The aim of this study was to quantify the release and the cytotoxicity of residual (co-)monomers and additives from PMMA-based splint materials under consideration of real splint sizes. Three different materials used for additive (3D printing), subtractive (milling) and conventional (powder and liquid) manufacturing were examined. METHODS: The splint materials SHERAprint-ortho plus (additive), SHERAeco-disc PM20 (subtractive) and SHERAORTHOMER (conventional) were analysed. 16 (n = 4) sample discs of each material (6 mm diameter and 2 mm height) were polished on the circular and one cross-section area and then eluted in both distilled water and methanol. The discs were incubated at 37 °C for 24 h or 72 h and subsequently analysed by gas chromatography/mass spectrometry (GC/MS) for specifying and quantifying released compounds. XTT-based cell viability assays with human gingival fibroblasts (HGFs) were performed for Tetrahydrofurfuryl methacrylate (THFMA), 1,4-Butylene glycol dimethacrylate (BDDMA) and Tripropylenglycol diacrylate (TPGDA). In order to project the disc size to actual splint sizes in a worst-case scenario, lower and upper jaw occlusal splints were designed and volumes and surfaces were measured. RESULTS: For SHERAeco-disc PM20 and for SHERAORTHOMER no elution was determined in water. SHERAprint-ortho plus eluted the highest THFMA concentration of 7.47 µmol/l ±2,77 µmol/l after 72 h in water. Six (co-)monomers and five additives were detected in the methanol eluates of all three materials tested. The XTT-based cell viability assays resulted in a EC50 of 3006 ± 408 µmol/l for THFMA, 2569.5 ± 308 µmol/l for BDDMA and 596.7 ± 88 µmol/l for TPGDA. SIGNIFICANCE: With the solvent methanol, released components from the investigated splint materials exceeded cytotoxic concentrations in HGFs calculated for a worst-case scenario in splint size. In the water eluates only the methacrylate THFMA could be determined from SHERAprint-ortho plus in concentrations below cytotoxic levels in HGFs.

Monomer release from surgical guide resins manufactured with different 3D printing devices.

OBJECTIVES: The aim of this study was to assess the post curing monomer release of resins applicable for 3D printing of surgical implant guides in dependency of printing technique and storing media using high performance liquidchromatography. MATERIAL AND METHODS: Specimens of Nextdent SG, Freeprint Splint, Fotodent Guide, 3Delta Guide, and V-print SG (n = 4) were additively manufactured with the corresponding DLP/SLA printing devices (Rapidshape D20II, Form2, Solflex350). Postprocessing was done according to the manufacturer's specifications. Subsequently, samples were eluted in methanol and water for 3 days and analyzed with gas chromatography/mass spectrometry (GC/MS). RESULTS: A total of twelve different substances released from the tested resin materials. The highest eluted concentration for MMA in methanol was 20.27 ± 8.60 µg/mL followed by 12.66 ± 3.38 µg/mL of HPMA. HEMA was found at concentration of 11.17 ± 2.43 µg/mL in methanol and 1.15 ± 0.11 µg/mL in water. TPGDA and TEGDMA reached maximum concentration in methanol of 4.29 ± 0.54 µg/mL and 5.07 ± 0.93 µg/mL and in water of 0.79 ± 0.19 µg/mL and 0.36 ± 0.14 µg/mL, respectively. Significant difference was found for the material Nextdent SG manufactured on SLA and DLP printing device for THFMA (p = 0.041), TEGDMA (p = 0.026), TPGDA (p = 0.05) and EGDMA (p = 0.06). The amount of monomers released into water did not reach the detection threshold for V-print SG. SIGNIFICANCE: The study revealed significant influence of the printing technique and resin material on the elution of monomers. The elution in methanol and water was significantly different. While the relative amount of eluted monomers from 3D printed guides is comparable to conventional direct composites and below toxic relevant concentrations, the absolute amount of monomer can rise in a clinic situation due to the size of the guides.

Actovegin® reduces PMA-induced inflammation on human cells.

PURPOSE: The effect of Actovegin® was investigated on PMA- and LPS-induced human peripheral blood mononuclear cells (PBMCs). METHODS: PBMCs (1 × 106 cells/ml) from five blood donors (2 f, 3 m; 45-55 years) were grown in medium and exposed to Actovegin® in the presence or absence of PMA or LPS. Supernatants were collected to assess the concentration of cytokines (TNF-α, IL-1beta, IL-6 and IL-10). The reactive oxygen species (ROS) were assessed by a ROS-GloTM H2O2 assay. RESULTS: Stimulation of cells by PMA or LPS (without Actovegin®) significantly increased the secretion of IL-1beta, IL-6, IL-10 and TNF-α from PBMCs, compared to controls. Pre-treatment of cells with Actovegin® (1, 5, 25, 125 µg/ml) plus PMA significantly decreased the secretion of IL-1beta from PBMCs, compared to controls (PMA without Actovegin®). In contrast, addition of Actovegin® (1, 5, 25, 125 and 250 µg/ml) plus LPS did not alter the IL-1beta production, compared to controls (LPS without Actovegin®). TNF-α, IL-6 and IL-10 do not contribute to the reduction of inflammatory reactions with Actovegin®. CONCLUSIONS: Actovegin® can reduce the PMA-induced IL-1beta release and the ROS production from PBMCs. These findings may help to explain the clinically known positive effects of Actovegin® on athletic injuries with inflammatory responses (e.g., muscle injuries, tendinopathies).

Titanium and zirconium release from titanium- and zirconia implants in mini pig maxillae and their toxicity in vitro.

OBJECTIVE: Titanium (Ti)- and Zirconia (ZrO2)-implants in mini pig maxillae were compared with respect to Ti/zirconium (Zr) release into the surrounding bone tissues, the resulting short term tissue responses and the potential toxicity. METHODS: Ti/Zr release from Ti- and ZrO2-implants in mini pig maxillae was determined with inductively coupled plasma optical emission spectrometry (ICP-OES) and inductively coupled plasma mass spectrometry (ICP-MS). The spatial distribution of Ti and Zr in maxilla tissues near the implant surface was assessed with laser ablation-inductively coupled plasma-mass spectrometry (LA-ICP-MS). A histological analysis was performed to investigate the tissue responses after 12 weeks of implantation. The cytotoxicity and DNA damage of Ti particles and ZrO2 particles were studied with XTT and Comet assay. RESULTS: The mean Ti content in the bone adjacent to Ti-implants was 1.67 mg/kg-bone weight. The highest Ti content detected was 2.17 mg/kg-bone weight. The mean Zr content in the bone adjected to ZrO2-implants was 0.59 mg/kg-bone weight. The highest Zr content was 0.75 mg/kg-bone weight. The spatial distribution of the Ti and Zr in bone showed mainly a higher intensity of Ti and Zr close to the screw thread outer tip rather. Histological analysis indicated that near both implant-types signs of bone marrow fibrosis were present. EC50 of commercially available ZrO2-nanoparticles (NPs, <100 nm) and ZrO2-microparticles (MPs, <5 µm) was 13.96 mg/ml and 80.99 mg/ml, respectively. ZrO2-NPs and ZrO2-MPs can induce DNA damage at 70 µg/ml and 810 µg/ml, respectively. SIGNIFICANCE: After 12-weeks of implantation, increased concentrations of Ti and Zr can be detected in bone/tissues near Ti- and ZrO2-implants in mini pig maxillae. Ti content released from Ti-implants is two times higher than the Zr content released from ZrO2-implants. ZrO2-NPs showed lower cytotoxicity and DNA damage compared to results reported for Ti-NPs in human cells.

The implanted glucose monitoring system Eversense: An alternative for diabetes patients with isobornyl acrylate allergy.

BACKGROUND: Some patients with diabetes develop skin reactions when using systems for continuous glucose monitoring (CGM) or insulin pumps. Regular usage and long wearing periods lead not only to skin irritation, but also to allergic contact dermatitis. It has been shown that allergens such as isobornyl acrylate (IBOA) are present in the plastic housing and also in the adhesives of medical devices used for diabetes treatment. OBJECTIVES: To evaluate the IBOA content of all parts of a newly introduced, implanted CGM system (Eversense) to check whether this can be an alternative for IBOA-sensitized patients. METHODS: The IBOA content of the implanted sensor itself (n = 3), the transmitter (n = 3), and two different types of adhesive (white adhesive [n = 4] and clear adhesive [n = 4]) was measured by gas chromatography/mass spectrometry. RESULTS: No IBOA was found in any part of this CGM system. CONCLUSIONS: Patients with an IBOA allergy may be able to use this implanted CGM system.

Evaluation of Isobornyl Acrylate Content in Medical Devices for Diabetes Treatment.

Background: Along with increased usage of continuous glucose monitors, flash glucose monitors, and patch pumps by patients with diabetes, the frequency of skin reactions has also increased. Skin irritation and itching can be annoying to users. However, more serious contact allergies to one or more components of the adhesives or plastic material of the housing of the devices can become lifelong. Redness and itchiness are so strong that patients can no longer use a particular system. In August 2017, a major culprit allergen, isobornyl acrylate (IBOA), was identified for these more serious reactions. Objectives: Our objective was to evaluate IBOA content in different medical products. Methods: The plastic material used for the housing of the Freestyle Libre (n = 3), Dexcom G6 (n = 3), and Enlite (n = 4) was analyzed for IBOA content by gas chromatography-mass spectrometry. Adhesives of the different systems were also analyzed. Results: IBOA was found in the housings of Freestyle Libre and Enlite sensor, but not in the Dexcom G6. Conclusions: Patients with an IBOA allergy should consider switching to a medical product without IBOA. Furthermore, removal of IBOA from devices that contact the skin is encouraged.

Wear of dental materials: Clinical significance and laboratory wear simulation methods -A review.

This review focusses on tribological aspects of teeth during function, the clinical significance of wear, wear of natural teeth and restorative materials and laboratory methods to simulate wear of restorative materials. Ceramic, metal alloy and amalgam show low material wear, whereas resin-based materials demonstrate substantial wear in the long term. The clinical wear shows a high variability with the patient factor accounts for about 50% of the variability. Wear as such seldomly compromises the function of the stomatognath system or individual teeth and is in most cases an esthetic problem. Particles that are ingested due to attrition and abrasion wear may pose a health risk to the patient, especially those from composite resin materials. However, systematic clinical studies on that issue are not available. For laboratory research many wear simulation devices and methods have been developed but only few are validated and have a moderate correlation with clinical wear.

Antioxidants as a novel dental resin-composite component: Effect on elution and degree of conversion.

OBJECTIVE: Ascorbic acid (Asc) and N-acetylcysteine (NAC) were reported to reduce genotoxicity induced by dental (co)monomers and their epoxy metabolites. The aim of the present study was to investigate Asc or NAC as novel components in light-curable methacrylate based dental composites regarding their effects on degree of conversion (DC) and elution of composite components. Additionally, the release of Asc or NAC was determined. METHODS: Asc or NAC (1, 0.1, 0.01 or 0 wt%) was experimentally incorporated into the composites Venus®, Grandio® and FiltekTM Supreme XTE and polymerized according to the instruction of manufacturers. The samples were elussted in methanol and water. For each composite-antioxidant mixture and elution medium four samples (n = 4) were prepared. The eluates were analyzed by gas chromatography/mass spectrometry (GC/MS), high-performance liquid chromatography/ultraviolett/diode array detection (HPLC/UV/DAD) and high-performance liquid chromatography/fluorescence detection (HPLC/FLD). DC of composite-antioxidant mixtures was measured in real-time with Fourier transform infrared spectroscopy (FTIR). RESULTS: The highest concentrations of eluted Asc were 313.98 µM (Venus®-1 wt% Asc; 1 day; methanol) and 245.34 µM (FiltekTM Supreme XTE-1 wt% Asc; 5 min; water). The highest concentrations of eluted NAC were 42.99 µM (1 day; Filtek™ Supreme XTE-1 wt% NAC; 1 day; methanol) and 108.11 µM (Filtek™ Supreme XTE-1 wt% NAC; 7 day; water). Triethylene glycol dimethacrylate (TEGDMA) elution was significantly increased in Venus®-1 wt% Asc and Grandio®-1 wt% Asc (1 day and 7 day methanol/water), compared to control. No significant difference was found for TEGDMA elution in Filtek™ Supreme XTE-1 wt% Asc/NAC. DC was significantly decreased compared to control (= composite without antioxidant) in Grandio® and Filtek™ Supreme XTE after 1, 0.1 and 0.01 wt% Asc incorporation and in Venus® after 1 and 0.1 wt% Asc incorporation. For composite-NAC mixtures, only DC of Grandio®-1 wt% NAC was significantly reduced. SIGNIFICANCE: Incorporation of NAC (1 wt%), as a novel composite component, into Filtek™ Supreme XTE, had no effect on DC and composite component elution, and supplies sufficient amount of antioxidant which may reduce toxicity. Therefore, it represents a beneficial mixture.

The Dexcom glucose monitoring system-An isobornyl acrylate-free alternative for diabetic patients.

BACKGROUND: Glucose monitoring systems, for example, Freestyle Libre (Abott) and Dexcom (Nintamed), are increasingly being used instead of conventional blood sugar measurement. However, many patients have experienced adverse skin reactions such as severe allergic contact dermatitis (ACD). Finally, in August 2017, the culprit allergen in Freestyle Libre, isobornyl acrylate (IBOA), was identified. OBJECTIVES: After patients have developed ACD, it is recommended that they no longer use their glucose monitoring systems. Thus, it is important to find an alternative IBOA-free device. PATIENTS AND METHODS: Five patients presented with ACD caused by Freestyle Libre. Each was patch tested with allergens from the baseline series and from a plastics and glues series, and additionally with IBOA 0.1% pet. Gas chromatography-mass spectrometry (GC/MS) of the Freestyle Libre sensor and the Dexcom sensor was performed. The Dexcom sensor remained on the skin of all patients for at least 2 days. RESULTS: All patients were sensitized to IBOA. GC/MS showed the presence of IBOA in the Freestyle Libre sensor, whereas the Dexcom sensor was IBOA-free. None of the patients had skin reactions to the Dexcom sensor. CONCLUSIONS: Patients with Freestyle Libre and IBOA allergy may use the Dexcom sensor as an alternative for glucose monitoring.

Effect of eluates from zirconia-modified glass ionomer cements on DNA double-stranded breaks in human gingival fibroblast cells.

OBJECTIVE: To formulate novel glass ionomer cements (GICs) containing zirconia (nanoparticles (NPs) and micro-particles (MPs)) and investigate the genotoxic effect of their eluates on DNA double-strand breaks of human gingival fibroblasts (HGFs) in vitro using a γ-H2AX fluorescent assay. METHODS: GIC (control, C), 10%ZrO2NPsGIC (T1) and 10%ZrO2MPsGIC (T2) were prepared per the manufacturer's instructions (hand-mixed, P/L=3.4:1w/w%). Dulbecco's modified Eagle's medium (DMEM) was used as the culture medium for HGFs and for eluate preparation. Eluates were collected from all specimens (n=5/g, 5×2mm) after 24h and used for XTT to obtain the EC50 using Graph Pad Prism4. A γ-H2AX immunofluorescence assay was performed to detect DSBs in HGFs. The mean foci per cells and percentage of free foci cells were statistically compared (one-way ANOVA with Tamhane's post hoc and Chi-square respectively) (p<0.05). RESULTS: (1) EC50 ranged from 31 to 36%. 5% and 20% eluate concentrations were selected for the genotoxicity test. (2) Cells exposed to eluates from T1 had lower mean foci per cell than cells in T2 and C eluates (p<0.05). Only cells in T1 at 5% had lower mean foci cell than medium (p<0.05). (3) T1 and C at both concentration showed a higher, but not significant, percentage of free foci cells than negative control (medium). At 20% eluate concentration T2 had a lower percentage of free foci cells than C (p<0.05). SIGNIFICANCE: Nano-zirconia GIC and micro-zirconia GIC were formulated. GIC and both zirconia modified GICs had no genotoxic effect on HGFs in vitro. Further studies related to physical properties should be performed to determine the future clinical applications for these novel nanomaterials.

Scientific update on nanoparticles in dentistry.

Nanoparticles having a size from 1 to 100 nm are present in nature and are successfully used in many products of daily life. In dental materials, nanoparticles are typically embedded but they may also exist as by-products from milling processes. Possible adverse effects of nanoparticles have gained increased interest, with the lungs being the main target organ. Exposure to nanoparticles in the dental laboratory is addressed by legal regulations. In dental practice, nanoparticles are mainly produced by intra-oral grinding/polishing and removal of materials, by wear of restorations or release from dental implants. Based on worst-case mass-based calculations, the additional risk as a result of exposure to nanoparticles is considered to be low. However, more research is needed, especially on vulnerable groups (patients with asthma or chronic obstructive pulmonary disease). An assessment of risks for the environment is not possible because of lack of data. Exposure-reduction measures mainly include avoidance of abrasive processes (for example, by proper sculpturing), cooling by the use of water spray and sufficient ventilation of treatment areas.

Induction of DNA double-strand breaks in human gingival fibroblasts by eluates from titanium dioxide modified glass ionomer cements.

OBJECTIVES: (1) To investigate the genotoxicity of a glass ionomer cement (GIC) and GIC incorporated with titanium dioxide nanopoarticle (TiO2NPs) and with microparticle (TiO2MPs) on DNA double-strand breaks of human gingival fibroblast cells (HGFs). (2) To compare the genotoxic differences between GIC and two modified cements. METHODS: TiO2NPsGIC and TiO2MPsGIC were prepared by adding 10% w/w of TiO2NPs and TiO2MPs to the GIC powder and hand-mixed followed the manufacturer instruction. Dulbecco's Minimum Essential Medium (DMEM) was used as a culture medium for HGFs and eluate preparation. Eluates from all groups were collected for XTT cell viability assay to obtain EC50 values. γ-H2AX immunofluorescence assay was performed to detect DNA double-strand breaks (DSBs) of HGFs. RESULTS: EC50 values were from 38% to 60% and eluate concentrations at 20% and 5% were selected for γ-H2AX immunofluorescence assay. At both concentrations, HGFs exposed to eluates from all cements groups had fewer mean foci per cell and higher percentage of free foci cells than H2O2 (p<0.05). At 20% concentration, cells exposed to eluates from both TiO2NPsGIC and TiO2MPsGIC groups had fewer mean foci per cell and higher percentage of free foci cell than GIC and culture medium (p<0.05). SIGNIFICANCE: Neither GIC nor 10% TiO2-modified GICs had a genotoxic effect on HGFs. Both TiO2NPsGIC and TiO2MPsGIC demonstrated less genotoxic effect than GIC. When comparing between the two modified cements, there was no genotoxic difference between the modified cements from different particle sizes (nanoparticle and micro-particle) of TiO2.

Bisphenol A release from an orthodontic resin composite: A GC/MS and LC/MS study.

OBJECTIVES: First, to analyse the in vitro release of BPA and Bis-GMA from an orthodontic resin composite (Transbond XT, 3M Unitek), stored in various conditions, by gas chromatography/mass spectrometry (GC/MS) and liquid chromatography/mass spectrometry (LC/MS); then to extrapolate the data to the clinical situation. Secondly, to explore the thermal stability of Bis-GMA. METHODS: Cylinders of resin composite were prepared and stored according to 3 different protocols: (1) they were light-cured 20s, then placed in artificial saliva; (2) they were light-cured 2s, then placed in acetonitrile; (3) they were light-cured 2s, then placed in methanol. For each group, BPA and Bis-GMA release were determined with GC/MS and/or LC/MS at least after one week. Besides, 120 brackets (10 of each type) were bonded over metal teeth, then debonded, and the weight and the surface of resin composite residues were measured. BPA and Bis-GMA release of adhesive residues were extrapolated from the data obtained with the cylinders. Besides, BPA release from a heated Bis-GMA solution was measured. RESULTS: With GC/MC, BPA was detected in all samples. With LC/MS, BPA was detected only from samples immersed in MeOH; Bis-GMA was detected, in varying amount according to the extraction media and the light-curing time. BPA was found after heating of the Bis-GMA solution. SIGNIFICANCE: Contamination risk and the heat applied in GC/MS may overestimate the BPA release from resin composite. Based on the LC/MS results, the risk of BPA release after orthodontic bonding would be more than 42000 times lower than the TDI for a 30-kg child.