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(1) Background: There is an ongoing discussion on the elution efficacy of antibiotic-impregnated cements. Our experiments were intended to clarify if there are differences in the antibiotic elution of HPLC compared with inhibition zone testing using eluates or PMMA discs. (2) Materials and Methods: Two cement brands with different concentrations of the active ingredient were tested in antimicrobial Kirby-Bauer (disc diffusion) assays. Cement platelets were directly applied on the agar plates and their zone of inhibition was measured. In parallel, the platelets were incubated in phosphate buffered saline (PBS) and at distinct points of time transferred into new buffer. At these time points, 50 µL of the bone cement eluates was used for zone of inhibition testing. Standard gentamicin sulfate solutions served as a control in the same test setup. To verify the microbiological investigations, the antibiotic content of the eluates was also measured via high-performance liquid chromatography (HPLC). (3) Results: The experiments with cement eluates showed better differentiable results than the direct application of the cement discs. The results were also comparable to investigations with HPLC and gentamicin sulfate standard solutions. (4) Conclusions: The results of elution rates are influenced by the test system and the period of observation chosen. The microbial test systems reflect the results of HPLC to the same degree and give evidence of the efficacy of the antibiotics. The HPLC tests on eluates were more suitable in representing differences in release characteristics.

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Background: This article presents a detailed analysis of a dataset consisting of 27 elements found in soils, soil eluates, and vegetables from private gardens in a region with a long history of coal mining and burning. With coal being one of the world's most significant energy sources, and previous studies highlighting elevated element levels in vegetables from this region, the objective of this study was to identify the factors that impact soil geochemistry and metal(loid) uptake in plants. Methods: Total major and trace element concentrations were analyzed in soils, soil eluates and vegetables by high resolution inductively coupled plasma mass spectrometry. The vegetable samples included six species: fennel, garlic, lettuce, parsley, onion, and radicchio. Each plant was divided into roots, stems, leaves, and/or bulbs and analyzed separately. In addition, the soil pollution status, bioavailable fractions and transfer factors from soil and soil eluates to different plant parts were determined. Results: The comprehensive dataset revealed that, apart from the substrate enriched with various elements (Al, As, Co, Cr, Mo, Ni, Pb, Sb, Sn, Ti, U, V, and Zn), other anthropogenic factors such as the legacy of coal mining and combustion activities, associated industries in the area, transport, and agricultural practices, also influence the elevated element concentrations (Cd, Cu, Fe, Mn, and Se) in locally grown vegetables. The transfer factors based on element concentrations in aqueous soil eluates and element bioavailable fractions confirmed to be an effective tool for evaluating metal uptake in plants, emphazising to some extent the effects of plant species and revealing unique patterns for each pollution source within its environmental context (e.g., Cd, Mo, S, and Se in this case). The study highlights the crucial importance of utilizing comprehensive datasets that encompass a multitude of factors when interpreting the impacts of element uptake in edible plants.

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OBJECTIVES: Various dental resin materials are available for the fabrication of temporary restorations using modern additive printing methods. Albeit these materials are placed for several months in intimate contact with dental hard and soft tissues, including the gingival crevice, there exists only insufficient evidence on the biocompatibility of these materials. This in vitro study aimed to delineate the biocompatibility of 3D printable materials on periodontal ligament cells (PDL-hTERTs). METHODS: Samples of four dental resin materials for additive fabrication of temporary restorations using 3D printing (MFH, Nextdent; GC Temp, GC; Freeprint temp, Detax; 3Delta temp, Deltamed), one material for subtractive fabrication (Grandio disc, Voco) and one conventional temporary material (Luxatemp, DMG) were prepared with a standardized size according to the manufacturer's instructions. Human PDL-hTERTs were exposed to resin specimens or eluates of the material for 1, 2, 3, 6 and 9 days. For determination of cell viability, XTT assays were performed. In addition, the expression of the proinflammatory cytokines interleukin 6 and 8 (IL-6 and 8) was assessed in the supernatants with ELISA. Cell viability and the expression of IL-6 and 8 in presence of the resin material or their eluates was compared with untreated controls. Immunofluorescence staining for IL-6 and IL-8, as well as scanning electron microscopy of the discs after culturing, were performed. Differences between groups were analyzed with Student´s t-test for unpaired samples. RESULTS: Compared to untreated control samples, the exposure against the resin specimen induced strong reduction of cell viability in case of the conventional material Luxatemp (p < 0.001) and the additive material 3Delta temp (p < 0.001) irrespective of the observation period. On the contrary, the presence of eluates of the various materials induced only minor changes in cell viability. Considering IL-6 (day 2: p = 0.001; day 6 and 9: p < 0.001) and IL-8 (day 1: p = 0.001; day 2, 3, 6, 9: p < 0.001) their expression was strongly reduced in presence of the eluate of Luxatemp. Except for IL-6 at day 1 and 6 also the material 3Delta temp caused significant reduction of both proinflammatory mediators at any time point. SIGNIFICANCE: The conventional material Luxatemp and the additive material 3Delta temp appear to severely affect cell viability when in direct contact with PDL-hTERTs. The other tested materials of this new category of additive materials and the subtractive material Grandio seem to induce only minor changes in direct contact with these cells. Therefore, they could serve as a viable alternative in the fabrication of temporary restorations.

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BACKGROUND: Safety issues for dental restorative composites are critical to material selection, but, limited information is available to dental practitioners. This study aimed to compare the chemical and biological characteristics of three nanohybrid dental composites by assessing filler particle analysis, monomer degree of conversion (DC), the composition of eluates, and cytotoxicity and reactive oxygen species (ROS) production in fibroblasts. METHODS: Three nanohybrid composites (TN, Tetric N-Ceram; CX, Ceram X Sphere Tec One; and DN, DenFil NX) were used. The size distribution and morphology of the filler particles were analysed using scanning electron microscopy (n = 5). The DC was measured via micro-Raman spectroscopy (n = 5). For the component analysis, methanol eluates from the light-polymerised composites were evaluated by gas chromatography/mass spectrometry (n = 3). The eluates were prepared from the polymerised composites after 24 h in a cell culture medium. A live/dead assay (n = 9) and Water-Soluble Tetrazolium-1 assay (n = 9) were performed and compared with negative and positive controls. The ROS in composites were compared with NC. Statistical significance in differences was assessed using a t-test and ANOVA (α = 0.05). RESULTS: Morphological variations in different-sized fillers were observed in the composites. The DC values were not significantly different among the composites. The amounts of 2-hydroxyethyl methacrylate (HEMA) were higher in TN than DN (p = 0.0022) and triethylene glycol dimethacrylate (TEGDMA) in CX was higher than in others (p < 0.0001). The lowest cell viability was shown in CX (p < 0.0001) and the highest ROS formation was detected in TN (p < 0.0001). CONCLUSIONS: Three nanohybrid dental composites exhibited various compositions of filler sizes and resin components, resulting in different levels of cytotoxicity and ROS production. Chemical compositions of dental composites can be considered with their biological impact on safety issues in the intraoral use of dental restorative composites. CX with the highest TEGDMA showed the highest cytotoxicity induced by ROS accumulation. DN with lower TEGDMA and HEMA presented the highest cell viability.

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OBJECTIVE: This study evaluated the cytotoxic and estrogenic effects of dust and eluates released into simulated wastewater after grinding of dental resin-based materials. METHODS: Four materials were used: ceram.x® universal, Filtek™ Supreme XTE, Lava™ Ultimate and Core-X™ flow. From each composite material, samples (5 × 2 mm, n = 50) were prepared according to the manufacturers' instructions. Lava™ Ultimate was used as blocks. All samples were ground to dust with a diamond bur (106 µm) and suspended in distilled water at 60 mg/mL. After storage for 72 h, the suspensions were separated into a soluble (eluate) and a particulate (dust) fraction. Eluates and dusts were evaluated for inhibition of Vibrio fischeri bioluminescence and cytotoxicity on human A549 lung cells (WST-1-Assay). The estrogenic activity was assessed by YES-Assay using Saccharomyces cerevisiae. Additionally, dental monomers (BisGMA, BisEMA, UDMA, TEGDMA, HEMA) and Bisphenol A were investigated. RESULTS: All eluates showed inhibition of V. fischeri bioluminescence at concentrations above 1.1 mg/mL (p < 0.05). The activity of the eluates of ceram.x® universal and Filtek™ Supreme XTE was significantly higher than Lava™ Ultimate and Core-X™ flow (p < 0.05). In the WST-1-Assay, all materials induced cytotoxic effects at concentrations of 0.1 mg/mL (p < 0.05), while no significant differences were detected among them. The tested materials revealed no estrogenic activity. All dental monomers and Bisphenol A showed concentration dependent cytotoxic effects (p < 0.05), whereas only Bisphenol A induced an estrogenic effect (p < 0.01). SIGNIFICANCE: Dust and eluates of resin-based dental materials released into wastewater exert bactericidal and cytotoxic effects in vitro. However, they reveal no estrogenic effect.

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This study evaluated the cytotoxicity of methacrylate-based resins containing dimethyl sulfoxide (DMSO). DMSO was incorporated into hydrophobic (R2) and hydrophilic (R5) resins at weight concentrations of 0, 0.01, 0.1, 1, 5, or 10 w/w %. Resin discs (n = 10/group) were prepared. Human gingival fibroblasts (HGF-1) were exposed to resin eluates for 24 h. Furthermore, dentin barrier test was performed using 3-D cultures of odontoblast-like cells (SV40 transfected pulp derived cells) with dentin slices of 400 µm thickness (n = 8). After acid etching of dentin, DMSO-modified resins were applied into the cavity part of the device and light-cured for 20 s. Cell viability (%) was assessed by MTT and analyzed spectrometrically. Data were analyzed by ANOVA and Tukey test (α = 0.05). Resin eluates showed statistically significantly lower % cell viability for all neat and DMSO-modified resins than seen for the negative control. Moreover, DMSO-R5 eluates resulted in significantly lower % cell viability than DMSO-R2 emulates. The dentin barrier test showed that DMSO-R2 did not result in significantly lower % cell viability, whereas incorporation of 1-10 w/w % DMSO into R5 resulted in significantly lower % of cell viability. Incorporating DMSO into hydrophilic self-etching resins may increase cytotoxicity. The biocompatibility is not influenced by the addition of DMSO into hydrophobic resin.

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A great number of different types of materials have been used in dentistry as intermediate restoratives. Among them, new resin-based bases have been released in the dental market. The present study focuses on the identification of the organic eluates released from such materials and the study of their surface microstructure in combination with their corresponding elemental composition. For this purpose, the following materials were used:ACTIVA™BioACTIVE-BASE/LINER™, Ketac™Bond Glass Ionomer, SDR™ and Vitrebond™Light Cure Glass Ionomer Liner/Base. Methanolic leachates derived from polymerized materials were analyzed by means of gas chromatography-mass spectrometry (GC-MS). Scanning electron microscopy(SEM) was used for the surface monitoring of suitably prepared specimens. The GC-MS analysis revealed the elution of twenty different substances from the three resin-based materials, while none was eluted from the glass ionomer base. The SEM analysis for Vitrebond™ presented small pits, the one for Ketac™Bond presented elongated cracks, while no voids were present for ACTIVA™BioACTIVE-BASE/LINER™ and SDR™. Moreover, the resin matrix of some dental materials may inhibit elements' accumulation on the surface layers. Particularly, the detected organic eluents may be related to potential toxic effects.

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A European round robin test according to ISO 5725-2 was conceptually prepared, realised, and evaluated. The aim was to determine the inter-laboratory variability of the overall process for the ecotoxicological characterization of construction products in eluates and bioassays. To this end, two construction products BAM-G1 (granulate) and HSR-2 (roof sealing sheet), both made of EPDM polymers (rubber), were selected. The granular construction product was eluted in a one stage batch test, the planar product in the Dynamic Surface Leaching test (DSLT). A total of 17 laboratories from 5 countries participated in the round robin test: Germany (12), Austria (2), Belgium (1), Czech Republic (1) and France (1). A test battery of four standardised ecotoxicity tests with algae, daphnia, luminescent bacteria and zebrafish eggs was used. As toxicity measures, EC50 and LID values were calculated. All tests, except the fish egg test, were basically able to demonstrate toxic effects and the level of toxicity. The reproducibility of test results depended on the test specimens and the test organisms. Generally, the variability of the EC50 or LID values increased with the overall level of toxicity. For the very toxic BAM-G1 eluate a relative high variability of CV = 73%-110% was observed for EC50 in all biotests, while for the less toxic HSR-2 eluate the reproducibility of EC50 varied with sensitivity: it was very good (CV = 9.3%) for the daphnia test with the lowest sensitivity, followed by the algae test (CV = 36.4%). The luminescent bacteria test, being the most sensitive bioassay for HSR-2 Eluate, showed the highest variability (CV = 74.8%). When considering the complex overall process the reproducibility of bioassays with eluates from construction products was acceptable.

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This study aimed to assess the potential impact on soil porewater, surface and groundwater from the beneficial application of organic wastes to soil, using their eluates and acute bioassays with aquatic organisms and plants: luminescence inhibition of Vibrio fischeri (15 and 30 min), Daphnia magna immobilization (48 h), Thamnocephalus platyurus survival (24 h), and seed germination of Lolium perenne (7 d) and Lactuca sativa (5 d). Some organic wastes' eluates promoted high toxic responses, but that toxicity could not be predicted by their chemical characterization, which is compulsory by regulatory documents. In fact, when organisms were exposed to the water-extractable chemical compounds of the organic wastes, the toxic responses were more connected to the degree of stabilization of the organic wastes, or to the treatment used to achieve that stabilization, than to their contaminant load. That is why the environmental risk assessment of the use of organic wastes as soil amendments should integrate bioassays with eluates, in order to correctly evaluate the effects of the most bioavailable fraction of all the chemical compounds, which can be difficult to predict from the characterization required in regulatory documents. According to our results, some rapid and standardized acute bioassays can be suggested to integrate a Tier 1 ecotoxicological evaluation of organic wastes with potential to be land applied, namely luminescence inhibition of V. fischeri, D. magna immobilization, and the germination of L. perenne and L. sativa.

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AIM: To investigate the cytotoxicity and genotoxicity of a new silicate-based BioRoot RCS® sealer in comparison with contemporary sealers. METHODOLOGY: A periodontal ligament cell line using lentiviral gene transfer of human telomerase reverse transcriptase (hTERT) was used and exposed to subtoxic concentrations of 24-h eluates from two epoxy resin-based (AH Plus Jet® and Acroseal® ), four various methacrylate-based endodontic sealers (EndoREZ® , RealSeal® , RealSeal SE® and Hybrid Root SEAL® ) and three silicate-based sealers (BioRoot RCS® , iRootSP® and MTA Fillapex® ). The XTT-based cell viability assay was used for cytotoxicity screening of materials. The γ-H2AX assay was used for genotoxicity screening. In the γ-H2AX immunofluorescence assay, PDL-hTERT cells were exposed to eluates of the substances for 6 h and DNA double-strand breaks (DSB) were detected microscopically. Induced foci represented DSBs, which can induce ATM-dependent phosphorylation of the histone H2AX. The statistical significance of the differences between the experimental groups was compared using the Student's t-test (P < 0.05). RESULTS: The cytotoxicity of the 24-h eluates could be ranked in the following order: Hybrid Root SEAL® >RealSeal® >Acroseal® >RealSeal SE® ≥ AH Plus Jet® > EndoREZ® >MTA Fillapex® > iRoot SP® >BioRoot RCS® . In negative controls (cells which received medium only) 4.08 ± 0.53 DSB foci (mean ± SEM) whilst in positive controls 10.76 ± 4.05 DSB foci/cell were found. BioRoot RCS® and RealSeal SE® exhibited significant differences in foci formation at 1/3 EC50 compared with their 1/10 EC50 concentration (P < 0.05). Both concentrations (1/10 and 1/3 of EC50) of AH Plus Jet® , Acroseal® , RealSeal® and MTA Fillapex® sealers were not significantly different when compared with the medium control (P < 0.05). CONCLUSIONS: New BioRoot RCS® was not toxic whilst Hybrid Root SEAL® demonstrated more toxicity and DNA double-strand breaks when compared with other resin- and silicate-based root canal sealers.

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BACKGROUND: Dried blood spots (DBS) are a useful specimen collection tool in situations where venous access is problematic, however, detection of biomarkers from this specimen type is subject to variation depending on storage conditions and storage time. OBJECTIVES: The objective of this study is to assess the detection of HBsAg, anti-HBc and anti-HCV from DBS after storage. STUDY DESIGN: DBS specimens were stored at -70°C, -20°C, 4°C, 22 to 28°C and 37°C either with or without desiccant. Eluates were also prepared from DBS specimens and stored at -20°C and -70°C. DBS cards and eluates were tested for HBsAg, anti-HBc and anti-HCV at baseline on day 0 and thereafter at intervals of 14, 70 and 200 days. RESULTS: Loss of detection of both HBsAg and anti-HBc was evident by the first time point (14 days) in all storage conditions except for the samples (DBS and eluates) stored at -20°C or -70°C. Both HBsAg and anti-HBc stored under these conditions showed minimal variation up to the final time point (200 days) of storage. The detection of anti-HCV did not differ between the 22 to 28°C, 4°C, -20°C and -70°C DBS nor the -20°C or the -70°C stored eluates over the 200 day time period. CONCLUSION: We suggest that extended storage of DBS intended for downstream testing is best carried out by freezing either the DBS, or eluate, at -20°C or -70°C as soon as possible following collection for optimal detection of HBsAg, anti-HBc and anti-HCV.

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BACKGROUND: The purpose of this study was to evaluate the release of chlorhexidine as an antifungal drug from doped self-cured poly (methyl methacrylate) (PMMA) acrylic resin and the effect of the drug released on the growth of Candida albicans. METHODS: Release of chlorhexidine was evaluated using liquid chromatography, and the effect of the drug on the growth of C. albicans was investigated microbiologically using a "well" technique on Saboraud culture medium inoculated with a resistant strain of C. albicans. RESULTS: Chlorhexidine leached steadily out of the acrylic resin into distilled water at mouth temperature, and the sustained drug release continued throughout the 28-day test period. The drug released also demonstrated antifungal activity against the resistant strain of C. albicans. CONCLUSION: The findings of this study support the use of chlorhexidine-impregnated self-cured PMMA chair-side resin as a new dosage form for the treatment of denture-induced stomatitis.

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OBJECTIVES: To monitor the release of the antifungal drugs Fluconazole, Chlorhexidine and a combination of the two from an auto-polymerized poly (methyl methacrylate) (PMMA) denture base resin; and to investigate the effect of the released drugs upon the growth of Candida albicans. METHODS: A high performance liquid chromatography-Ultra violet (HPLC-UV) method was used in the analysis of the released drugs into distilled water from PMMA discs doped with the antifungal drugs Fluconazole (10%), Chlorhexidine (10%) and a combination of the two drugs (5% each). The antifungal efficacy of the released drugs was monitored, microbiologically, employing "well" technique on a Saborauds culture medium inoculated with a resistant strain of Candida albicans. RESULTS: It was shown that Fluconazole, Chlorhexidine and the combination of the two drugs can be successfully incorporated with PMMA. It was found that the drugs leach steadily out of the PMMA resin into distilled water at mouth temperature and that sustained drug release continued throughout the 28 days test period. It was also shown that the released drugs demonstrated an antifungal activity against the resistant Candida albicans and this was most remarkable in the combined drugs samples. CONCLUSIONS: The findings of this investigation have a clinical value in terms of their significant contribution to the treatment of fungal infections of the oral cavity. The sustained release of anti-fungal drugs from the PMMA resin clearly constitutes a new dosage form of these drugs via the poly (methyl methacrylate) delivery system.