Cytotoxicity effects and differentiation potential of ormocer-based and nanohybrid composite resins on human dental pulp stem cells.

OBJECTIVE: to compare conventional nanohybrid (Ceram.x Spectra) and ormocer-based (Admira fusion) dental composite resins effects on human dental pulp stem cells (hDPSCs) in terms of cytotoxicity, self-renewal, migration and osteogenic differentiation. METHODS: hDPSCs were cultured in presence of different dilutions (undiluted, form 1:2 to 1:100) of CeramX (CX) and Admira fusion (AD) eluates and viability assay in standard or osteogenic conditions were performed. Samples and eluates were prepared according to ISO 10993-12. In addition, apoptosis, self-renewal and migration activity evaluations were carried out. Osteogenic differentiation potential was tested by Alkaline Phosphatase Activity, alizarin red staining and gene expression of specific markers (ALP, RUNX2, OCN, OPN and COL1α1). Statistical analysis was performed by means of a One-way analysis of variance (One-way ANOVA) followed by a Tukey's test for multiple comparison; results were presented as mean ± standard error of mean (SEM). RESULTS: Admira Fusion demonstrated to be highly biocompatible and showed positive effects on hDPSCs proliferation and differentiation; on the contrary, conventional nanohybrid composite showed to be more cytotoxic and without any notable effect on stem cells differentiation. Moreover, the obtained results were further corroborated by a significant upregulation of osteogenic differentiation markers obtained in presence of ormocer-based composite resin eluate. Specifically, in AD 1:50 group expression levels of ALP, Runx2, Col1α1 were double than control (ALP, p = 0.045; Runx2, p = 0.003; Col1α1, p = 0.001) and CX 1:50 (ALP, p = 0.006; RUNX2, p = 0.029; Col1α1, p = 0.005). Moreover, in the same group, OPN and OCN resulted about 5 times more expressed as compared to control (OPN, p = 0.009; OCN, p = 0.0005) and CX 1:50 (OPN, p = 0.012; OCN, p = 0.0006). SIGNIFICANCE: The less cytotoxicity obtained by AD than conventional nanohybrid composite may be attributed to a reduced monomers release in the oral environment, supporting the hypothesis of limited adverse effect and enhanced healing potential, mainly when the material is positioned in close contact with pulp tissue.

The influence of copper-doped mesoporous bioactive nanospheres on the temperature rise during polymerization, polymer cross-linking density, monomer release and embryotoxicity of dental composites.

OBJECTIVES: Composites with copper-doped mesoporous bioactive nanospheres (Cu-MBGN) were developed to prevent secondary caries by imparting antimicrobial and ion-releasing/remineralizing properties. METHODS: Seven experimental composites containing 1, 5 or 10 wt% Cu-MBGN, the corresponding inert controls (silica) and bioactive controls (bioactive glass 45S5) were prepared. The temperature rise during light curing, cross-linking density by ethanol softening test, monomer elution and their potential adverse effects on the early development of zebrafish Danio rerio was investigated. RESULTS: Materials combining Cu-MBGN and silica showed the highest resistance to ethanol softening, as did the bioactive controls. Cu-MBGN composites showed significant temperature rise and reached maximum temperature in the shortest time. Bisphenol A was not detected, while bis-GMA was found only in the control materials and TEGDMA in the eluates of all materials. There was no increase in zebrafish mortality and abnormality rates during exposure to the eluates of any of the materials. CONCLUSIONS: The composite with 5 wt% Cu-MBGN combined with nanosilica fillers showed the lowest ethanol softening, indicating the polymer's highest durability and cross-linking density. Despite the TEGDMA released from all tested materials, no embryotoxic effect was observed.

Biocompatibility and acid resistance of preformed crowns in children: an in vitro study.

PURPOSE: To investigate the in vitro biocompatibility of human gingival fibroblasts with preformed paediatric crowns and resistance to acid exposure at levels that simulate the oral environment. METHODS: This laboratory study investigated primary HGFs viability, metabolic activity, cytotoxicity, and apoptotic events on preformed metal crown discs, composite resin-coated wells, and monolithic zirconia fragments at 24, 48, and 72 h using the ApoTox-Glo Triplex assay. The PPCs were also immersed in 0.1% lactic acid, 0.2% phosphoric acid, or 10% citric acid for 7 days at 37 °C to reproduce conditions associated with dietary intake or gastric reflux. Samples were then subject to inductively coupled plasma optical emission spectrometry to quantitate the release of ions. RESULTS: The viability of HGFs on stainless steel and CR significantly declined at 48 and 72 h, representing potential cytotoxicity (p < 0.05). Cytotoxicity of HGFs was also higher for stainless steel and ZR compared to control (p < 0.05). PMCs and ZR crowns gave minimal ion release. Meanwhile, significant quantities of metallic ions, including copper (Cu), iron (Fe), nickel (Ni), and zinc (Zn), were present in eluates from veneered-preformed metal crowns. CONCLUSION: As PPCs can be exposed to highly acidic environments for many years, thus the release of metallic ions from V-PMCs should form the further investigation in future studies.

Biocompatibility of bulk-fill resins in vitro.

OBJECTIVES: This study aims to evaluate the cytotoxicity and genotoxicity of three different extracts obtained from Filtek™ One Bulk Fill, Tetric Evoceram® Bulk Fill and Coltene Fill-Up! resins. MATERIALS AND METHODS: The cytotoxicity was determined on 3T3 fibroblast cells using the MTT and crystal violet assays. The genotoxicity was determined using a cytokinesis-block micronucleus assay. RESULTS: The cytotoxicity of the resin extracts on 3T3 mouse fibroblasts was found to be dose-dependent with both the MTT and crystal violet assays. Extracts concentrated above 1% were cytotoxic according to the MTT assay. The Filtek™ One Bulk Fill, Tetric Evoceram® Bulk Fill, and Coltene Fill-Up! resins reached the LD50 at concentrations of 60%, 50%, and 20%, respectively, and showed genotoxicity rates that were 2-5 times, 3-8 times, and 4-15 times higher than the negative control, respectively. CONCLUSIONS: Coltene Fill-Up! resin extracts were the most cytotoxic and genotoxic, followed by Tetric Evoceram® Bulk Fill and Filtek™ One Bulk Fill. CLINICAL RELEVANCE: The analyzed bulk-fill resins showed differences in in vitro biocompatibility, and the Filtek™ One Bulk Fill was found to be the safest for clinical use.

Comparison of Cytotoxicity and Genotoxicity in Three Types of Indirect Restorative Materials on Human Periodontal Stem Cells.

PURPOSE: This study aimed to compare the cell toxicity and biological characteristics of Ketac GIC (glass-ionomer cement), Nexus RMGIC (resin-modified glass-ionomer cement), and RelyX RC (resin cement) in human periodontal stem cells (PDSCs). MATERIALS AND METHODS: To compare the effects of Ketac GIC, Nexus RMGIC, and RelyX RC on PDSCs, the cements were diluted from 1:2 to 1:8. PDSCs were then treated with the serially diluted cements with or without N-acetyl-cysteine (NAC), and cell survival was measured using water-soluble tetrazolium salt (WST-1) assay. Intracellular reactive oxygen species (ROS) was measured using 2',7'-dichlorofluorescin diacetate (DCFDA), and western blot analysis was performed to observe phosphorylation and activation of extracellular signal-regulated kinase (ERK) by Nexus RMGIC or RelyX RC. RESULTS: Cell death and proliferation were dose-dependently reduced following Nexus RMGIC or RelyX RC treatment. In addition, Nexus RMGIC or RelyX RC showed an increase intracellular ROS generation compared to Ketac GIC. Pretreatment with NAC confirmed the suppression of cell toxicity and ROS generation induced by Nexus RMGIC or RelyX RC. Nexus RMGIC or RelyX RC activates ERK phosphorylation, not p38 phosphorylation, in PDSCs. CONCLUSION: This study showed that the treatment with Nexus RMGIC or RelyX generates intracellular ROS and cell death through the ERK signaling pathway in PDSCs. In contrast, these effects were not observed with Ketac GIC, indicating that resin-based materials may have cytotoxic and genotoxic effects on PDSCs.

Effect of thickness on the degree of conversion, monomer elution, depth of cure and cytotoxicity of bulk-fill composites.

PURPOSE: Today, bulk-fill composites are used as a single layer with a thickness of up to 4-5 mm. However, is proper polymerization achieved with this increased thickness? METHODS: This study was designed to investigate the effect of thickness on the degree of conversion (DC) (n = 6), the elution of monomers (n = 6), depth of cure (DoC) (n = 10) and cytotoxicity (n = 6) of the bulk-fill composites SDR Flow Plus (SDR), SonicFill2 SingleFill (SF) and ACTIVA Bioactive Restorative (ACT) in comparison to the conventional G-aenial Posterior (GC). Two-way analysis of variance (ANOVA) was used to assess the interaction between materials and surfaces, and one-way ANOVA and Tukey tests were used to compare the degree of conversion, monomer elution and cytotoxicity values (P < 0.05). RESULTS: The highest DC was found at the top surface of SDR, while the lowest DC was found at SF. The V2 mm/V0 mm DoC ratios of the composites except ACTs were appropriate according to the threshold. None of the composites were cytotoxic on day 1. CONCLUSION: In bulk-fill composites, DC decreased and monomer elution increased with increasing depth. The V4 mm/V0 mm ratios of all bulk-fill groups were not appropriate. Additionally, only ACTs had a cell viability of <70% on day 7.

Cytotoxicity and reactive oxygen species production induced by different co-monomer eluted from nanohybrid dental composites.

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.

Effects of nanohybrid flowable resin-based composites on fibroblast viability using different light-curing units.

PURPOSE: To investigate the in vitro cytotoxic effects of Bis-GMA-containing and Bis-GMA-free flowable resin-based composites (RBCs) on primary human gingival fibroblast cells (hGFc) using direct and indirect curing methods and three different light-curing units (LCUs). MATERIALS AND METHODS: Cells were isolated and cultured in vitro in 24-well plates. The plates were divided into treatment (cells with RBC), control (cells only), and blank (media only) groups. In the treatment groups, two types of nanohybrid flowable RBCs were used: Bis-GMA-free and Bis-GMA groups. Each treatment group was subdivided according to the curing method, i.e., direct curing (RBC was injected into the wells and cured directly on the attached cells) and indirect curing (the samples were pre-cured outside of the well plate and then added to the well plate with cells). To vary the LCU, the subgroups were further divided into three groups: multiple-emission peak light-emitting diode, single-emission peak light-emitting diode, and quartz-tungsten-halogen units. Curing was conducted for 20 seconds. The hGFc cytotoxicity was evaluated via 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) assay after 24, 48, and 72 hours of culturing. RESULTS: The MTT assay results showed that both RBCs were significantly cytotoxic toward hGFc compared to the control group (p < 0.0001). The Bis-GMA group was significantly more cytotoxic to the cells compared to the Bis-GMA-free group. In addition, the curing method and time interval affected cell viability regardless of the LCU used. CONCLUSION: The Bis-GMA flowable RBC and direct curing method had the highest cytotoxic effects on hGFc regardless of the LCU used. Careful selection of flowable RBCs and proper curing techniques are required to decrease the cytotoxic effects on hGFc and improve the clinical handling of oral tissues.

Influence of the self-adhering strategy on microhardness, sorption, solubility, color stability, and cytotoxicity compared to bulk-fill and conventional resin composites.

OBJECTIVES: To analyze and compare, in vitro, the microhardness, sorption, solubility, color stability, and cytotoxicity of three types of resin composites: self-adhesive (SARC) (Dyad Flow (DF)/Kerr), bulk-fill (Filtek Bulk Fill Flow (FBF)/3 M ESPE), and conventional (Filtek Z350XT Flow (Z350)/3 M ESPE). MATERIALS AND METHODS: Thirty cylindrical specimens were prepared using a split metal mold (15 mm × 1 mm), divided into 3 groups (n = 10) according to the material used. Vickers hardness (VH) was calculated from three indentations (300gf/15 s) per specimen. The sorption and solubility were measured according to the ISO 4049:2009 specification after storing in distilled water for 7 days. The color of each resin composite was measured using a portable digital spectrophotometer according to the CIELAB system. After a 7-day immersion in coffee, the color variation (∆E) was calculated. Following the ISO 10993:2012, the cytotoxicity in Vero cells was evaluated through the MTT assay. The results were analyzed using the Kruskal-Wallis test to compare the studied groups. The Wilcoxon test was used to compare the assessments in each studied group. For cytotoxicity analysis, the data were compared by the ANOVA test (α = 0.05). RESULTS: DF showed the lowest VH (28.67), highest sorption (0.543 µg/mm3) and solubility (1.700 µg/mm3), and higher ∆E after 7 days of coffee immersion (p = 0.008). The resin composites studied were considered non-cytotoxic. CONCLUSIONS: The SARC presented inferior mechanical and physical-chemical properties than bulk-fill and conventional resin composites, with comparable cytotoxicity against Vero cells. CLINICAL RELEVANCE: The simplification of the clinical protocol of SARC can minimize the number of possible failures during the restorative technique. However, considering their inferior physical and mechanical properties, their coverage with materials of higher mechanical properties and physical-chemical stability should be considered.

Characterization of universal chromatic resin-based composites in terms of cell toxicity and viscoelastic behavior.

OBJECTIVES: A current trend to simplify dental restorative procedures is toward using universal chromatic light-cured resin-based composites (RBCs) designed to adapt esthetically to various clinical situations. This study offers a comparative characterization of the mechanical and cytotoxic behavior of such materials that use different techniques to adjust their optical properties (e.g., structural color instead of pigment addition), have different filler systems but are based on a comparable organic matrix. METHODS: The structural appearance of the filler systems was assessed by scanning electron microscopy. Various quasi-static and viscoelastic parameters were evaluated at clinically relevant frequencies (0.5-5 Hz) using an instrumented indentation test with a Dynamic Mechanical Analysis (DMA) module. Cytotoxicity on human gingival fibroblasts (HGF-1), when exposed to eluates from tested RBCs specimens (up to one month), was assessed using a WST-1 colorimetric proliferation assay. Multifactor analysis of variance was applied to compare the parameters of interest (Martens, Vickers, and indentation hardness; elastic and total indentation work; creep, indentation depth; storage, loss, and indentation moduli; loss factor; cell viability) between analyzed RBCs, loading frequencies, and eluate age. RESULTS: Structural particularities of the filler systems are directly reflected in the mechanical behavior of the analyzed materials. Changes in the filler system, necessary to achieve structural color, generally resulted in lower mechanical properties but a better ability to absorb shock. In contrast, the cytotoxicity was comparable. SIGNIFICANCE: Based on the performed characterization, universal chromatic RBCs fits in the conventional RBCs class to expect comparable clinical behavior.

Effect of Finishing-Polishing Procedures on Cytotoxicity of Resin-Based Restorative Materials via Real-Time Cell Analysis.

OBJECTIVE: The aim of this study was to evaluate the effect of finishing and polishing procedures of compomer and bulk-fill composite resins on cytotoxicity against human gingival fibroblasts by xCELLigence analysis. STUDY DESIGN: Filtek™ Bulk Fill composite and Dyract XP compomer were used. After curing, the specimens were randomly divided into two groups and finishing-polishing procedures were applied to one group; no finishing-polishing procedures were applied to the other group. For the first time in this study, pure gold samples were prepared with the same weight and base area as the test specimens and the wells containing the pure gold samples were determined as the control group. xCELLigence system was used to assess the response of the human gingival fibroblasts after exposure to test specimens. Measurements were recorded for 72 hours after adding specimens. RESULTS: Finishing and polishing procedures caused a significant increase in cell viability of Dyract XP compomer samples at all time periods; the percentage of cell viability reached above 70% after finishing and polishing procedures. However, significant effects were not observed in Filtek™ Bulk Fill composite samples at any time period. CONCLUSION: Finishing and polishing procedures play an essential role in increasing the biocompatibility of Dyract XP compomer. It is recommended to apply finishing and polishing procedures even though a smooth surface may be obtained in restorations with matrix strips.

In vitro biocompatibility testing of different base materials used for elevation of proximal subgingival margins using human gingival epithelial cells.

PURPOSE: To analyze the biological effects of four base materials used for elevation of proximal subgingival margins on gingival epithelial cells. METHODS: Twenty-eight specimens for each of the four base materials (total 112 specimens) were used: resin-modified glass ionomer (RMGI), glass hybrid (HV-GIC), flowable bulk fill resin composite (Bulk Flow) and bioactive ionic resin (Activa). Proximal enamel and root dentin were used as controls. Gingival epithelial cell viability was calculated after direct incubation on all four types of material for either 24 h or 72 h using both the methyl tetrazolium and trypan blue dye exclusion assays. Data were analyzed statistically using one-way analysis of variance, Tukey post hoc test and independent sample t-test (P < 0.05). RESULTS: Cell viability values in both assays showed significant differences among the study groups. Bulk Flow showed the highest values, followed in order by Activa and the control groups. Both HV-GIC and RMGI had the lowest values. Cell viability in all of the study groups was higher after incubation for 72 h than after 24 h. CONCLUSION: In terms of biocompatibility with epithelial tissues, bulk fill resin composite appears to be most suitable, followed by bioactive composite, for subgingival placement than glass ionomer-based materials, especially that containing 2-hydroxy-ethyl methacrylate.

Physicochemical and biological evaluation of a triazine-methacrylate monomer into a dental resin.

OBJECTIVES: This study aimed to (1) formulate blend resins with 2.5 or 5 wt.% of the methacrylate monomer 1,3,5-triacryloylhexahydro-1,3,5-triazine (TAT), and (2) to evaluate the blend resins regarding the physicochemical and biological properties. METHODS: The base resin was formulated mixing 60 wt.% of bisphenol A glycol dimethacrylate and 40 wt.% of triethylene glycol dimethacrylate with photoinitiator/co/initiator system. TAT was added at 2.5 (G2.5%) or 5 (G5%) wt.%, and a group without TAT was used as control (Gctrl). The resins were analyzed for degree of conversion (DC), Knoop hardness (KHN), softening in solvent (ΔKHN), ultimate tensile strength (UTS), contact angle, surface free energy (SFE), antibacterial activity against Streptococcus mutans biofilm formation, and cytotoxicity against human keratinocytes. RESULTS: There was no difference for the DC (p = 0.676). The addition of TAT at 5 wt.% induced higher KHN (p<0.001), higher resistance against softening in solvent (p<0.001), and higher UTS (p = 0.04). There were no statistically significant differences for contact angle with water (p = 0.106), α-bromonaphtalene (p = 0.454), and SFE (p = 0.172). The higher the TAT concentration, the higher the antibacterial activity (p<0.001). G2.5% showed no cytotoxicity compared to Gctrl (p>0.05), and G5% induced lower cell viability (p<0.05). CONCLUSIONS: The addition of 2.5 wt.% of TAT is suitable for conveying antibacterial activity for dental resins without changing the physicochemical properties or impairing the cytotoxic effect. CLINICAL RELEVANCE: Methacrylate monomers that decrease bacterial viability and copolymerize with the resin matrix are exciting approaches to developing therapeutic materials. TAT showed promising properties to may hamper and prevent carious lesions when incorporated into dental materials. Further evaluations with higher cariogenic challenges will be carried to analyze the formulated materials.

Impact of ultra-fast (3 s) light-cure on cell toxicity and viscoelastic behavior in a dental resin-based composite with RAFT-mediated polymerization.

OBJECTIVE: The aim of the study was to determine the effects of ultra-fast (3 s) light-curing on the viscoelastic behaviour at clinically relevant frequencies, and cell toxicity, in a resin-based composite (RBC) with reversible addition-fragmentation-chain transfer (RAFT) mediated polymerization. METHODS: Three different protocols were used to cure cylindrical samples (height = 4 mm, Ï´ = 5 mm), including ultra-fast (3s) cure with high radiant emittance, 10 s and 20 s cure with moderate radiant emittance. The properties of the light curing device were evaluated in all curing protocols by spectrophotometry up to an exposure distance of 10 mm. The light transmission through the samples was determined in real-time with the same spectrophotometer. Absorbance was calculated as a function of wavelength. The quasi-static (indentation hardness/HIT, indentation modulus/EIT) and viscoelastic (storage modulus/E', loss modulus/E″, loss factor/tan Î´) material behavior was determined in an instrumented indentation test with a DMA (Dynamic Mechanical Analysis) module for 10 frequencies (0.5-5 Hz) by profiling the center of the samples in 330 µm steps from top to bottom. Cellular toxicity on human gingival fibroblast (HGF-1) was assessed using a WST-1 colorimetric assay after incubation time of up to 3 months. One and multiple-way analysis of variance (ANOVA) with Tukey honestly significant difference (HSD) post-hoc tests (α = 0.05) were applied. RESULTS: The irradiance transmitted through a 4 mm high sample was less than 7% of the incident irradiance, and the absorbance was similar for all curing protocols, showing a decrease with wavelength. Similar quasi-static and viscoelastic parameters were observed regardless of the curing protocol. HIT increased slightly and EIT, E', E″ and tan Î´ decreased with frequency. Occasionally, slightly higher confidence intervals were observed for the ultra-fast curing group, which were related to a potential accumulation of stress. The curing protocol had no effect on cell viability (p = 0.326) but the eluate age (p < 0.001, ηP2 = 0.879) did. None of the groups showed cell toxicity at any point in time with respect to the corresponding negative control. CONCLUSIONS: The ultra-fast curing with high irradiance induced no cell toxicity and an equivalent viscoelastic behavior as with conventional curing protocols in a RAFT-modified RBC.

Cytotoxicity and estrogenicity in simulated dental wastewater after grinding of resin-based materials.

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.

The effect of new anti-adhesive and antibacterial dental resin filling materials on gingival fibroblasts.

OBJECTIVE: Aim of this study was to evaluate the biocompatibility of four experimental antiadhesive and antibacterial dental filling composites on human gingival fibroblasts (HGFs). METHODS: For these experimental resin composites a delivery system based on novel polymeric hollow beads, loaded with Tego Protect (Aa1), Dimethicone (Aa2), Irgasan (Ab1) and methacrylated polymerizable Irgasan (Ab2) as active agents was used. The cultured HGFs' cell integrity, proliferation, viability, collagen synthesis and cytokine release were measured. For this purpose, human gingival fibroblasts were treated with eluates from all four composites and compared with an experimental standard composite (ST). Eluate extraction times 24 h and 168 h were chosen. RESULTS: Statistical analysis was conducted via a mixed model. Both antibacterial composites reduced proliferation, collagen and cytokine synthesis significantly (p < 0.05), increasing with time of elution. Ab1 did also have a damaging effect on the membrane and on cell viability. SIGNIFICANCE: Overall, it can be concluded that the antiadhesive composites showed clear advantages over the antibacterial composites in terms of biocompatibility. This study also continues to show the potential of the new poly-pore system, as it can be used for a variety of other applications in future composite mixtures.

The cytotoxic and oxidative effects of restorative materials in cultured human gingival fibroblasts.

The aim of this study was to evaluate the cytotoxic and oxidative effects of the most commonly used dental restorative materials on human gingival fibroblast cells (HGFCs). HGFCs were obtained from healthy individuals. The tested restorative materials were a microhybrid resin based composite, a compomer resin, a glass ionomer cement, and an amalgam alloy. One hundred eight cylindirical samples, 10 mm in diameter and 2 mm in height, were prepared according to ISO 10993-12:2002 specifications (n = 9 in the tested subgroups). Freshly prepared and aged samples in artificial saliva at 37 °C (7 and 21 d) were placed into well plates and incubated. Wells without dental materials were constituted as the control group. After 72 h incubation period, cytotoxicity was determined using the neutral red (NR) assay. Oxidative alterations were assessed using total antioxidant capacity (TAC) and total oxidant status (TOS) assay kits. Data were analyzed using the ANOVA and LSD post hoc tests. All tested materials led to significant decreases in the cell viability rates (33-73%) compared to the control group. Glass ionomer and resin composite were found to be more cytotoxic than amalgam alloy and compomer. The highest TAC level was observed in glass ionomer after seven-day aging and these changes prevented an increase in TOS levels. Increases in TAC levels after seven-day aging in all groups exhibited significant differences with freshly prepared samples (p < 0.05). In all material groups, TOS levels of freshly prepared samples differed statistically and significantly from samples aged for 7 and 21 d (p < 0.05). The data obtained suggested that all the tested materials exhibited cytotoxic and pro-oxidant features. Freshly prepared samples caused higher TOS levels. However, oxidant status induced by materials decreased over time.

Cytotoxicity evaluation of dental and orthodontic light-cured composite resins.

INTRODUCTION: The aim of this study was to determine the cytotoxicity of light-cured composite resins (Clearfil ES-2, Clearfil ES Flow, Filtek Supreme XTE, Grengloo, Blugloo, Transbond XT, and Transbond LR) then to assess leachable components in contact with human gingival fibroblasts (GFs) and to quantity detected bisphenol A (BPA). METHODS: Light-cured composite resin discs were immersed for 24 hours in gingival fibroblastic medium (n = 3 for each product) and in control medium (n = 2 for each product) contained in plate. Cytotoxicity of the products (n = 95) was determined by the measure of cell viability using MTT assay after reading the optical densities of the plates. The analysis of leachable components was done by gas phase chromatography and mass spectrometry (GC-MS) and detected BPA was quantified. The limit of quantification was 0.01 µg/mL. Statistical analyses were performed by using IBM SPSS Statistics 20 and Kruskal-Wallis and Mann-Whitney U-tests were applied. RESULTS: Cell viabilities were between 85 and 90%. Many chemical compounds including triethylene glycol dimethacrylate (TEGDMA) and BPA were identified. The average concentrations were 0.67 µg/mL ± 0.84 in the control medium and 0.73 µg/mL ± 1.05 in the fibroblastic medium. Filtek Supreme XTE presented the highest concentration of BPA with 2.16 µg/mL ± 0.65 and Clearfil ES Flow presented the lowest with 0.25 µg/mL ± 0.35. No BPA was detected with Transbond XT and Transbond LR. Clearfil ES Flow, Filtek Supreme XTE, Grengloo and Transbond LR presented residual TEGDMA. CONCLUSIONS: Light-cured composite resins are slightly cytotoxic opposite GFs and release many components including BPA and TEGDMA. Clinical precautions should be taken to decrease the release of these monomers.

Thiourethane-functionalized fillers: biological properties and degradation resistance

Abstract This study determined the effect of thiourethane-functionalized fillers (TU) on the antimicrobial properties, cytotoxicity, degree of conversion (DC), water sorption (Wsp) and solubility (Wsl) of experimental composites. TU-modified fillers were added at different ratios in experimental composites: 0 (Control-TU0), 25% (TU25), 50% (TU50), 75% (TU75) and 100wt% (TU100). The antimicrobial properties were detected through the exhaustion test and counting of Streptococus mutans colonies for biofilm formation. Cytotoxicity to human gingival fibroblasts was evaluated in three different parameters: XTT (2,3-Bis-(2-Methoxy-4-Nitro-5-Sulfophenyl)-2H-Tetrazolium-5-Carboxanilide), NRU (Neutral Red Uptake assay) and CVDE (Crystal Violet Dye Exclusion test)) at the same cells. ELISA was used to measure the IL-6 and b-FGF biomarkers. DC was determined by Fourier-transformed infrared spectroscopy, while Wsp and Wsl by mass variations. Inhibitory capacity of biofilm formation was not observed for any material. All groups presented at least 70% of cell survival within the observed periods (24h and 7 days). Positive control (toxic) had high IL-6 values and low b-FGF values. No significant variations in DC, Wsp, and Wsl were observed among the experimental groups. The use of thiourethane did not present antimicrobial and cytotoxic activity and the tested materials presented equivalent properties to those conventionally used in dentistry.

Thiourethane-functionalized fillers: biological properties and degradation resistance.

This study determined the effect of thiourethane-functionalized fillers (TU) on the antimicrobial properties, cytotoxicity, degree of conversion (DC), water sorption (Wsp) and solubility (Wsl) of experimental composites. TU-modified fillers were added at different ratios in experimental composites: 0 (Control-TU0), 25% (TU25), 50% (TU50), 75% (TU75) and 100wt% (TU100). The antimicrobial properties were detected through the exhaustion test and counting of Streptococus mutans colonies for biofilm formation. Cytotoxicity to human gingival fibroblasts was evaluated in three different parameters: XTT (2,3-Bis-(2-Methoxy-4-Nitro-5-Sulfophenyl)-2H-Tetrazolium-5-Carboxanilide), NRU (Neutral Red Uptake assay) and CVDE (Crystal Violet Dye Exclusion test)) at the same cells. ELISA was used to measure the IL-6 and b-FGF biomarkers. DC was determined by Fourier-transformed infrared spectroscopy, while Wsp and Wsl by mass variations. Inhibitory capacity of biofilm formation was not observed for any material. All groups presented at least 70% of cell survival within the observed periods (24h and 7 days). Positive control (toxic) had high IL-6 values and low b-FGF values. No significant variations in DC, Wsp, and Wsl were observed among the experimental groups. The use of thiourethane did not present antimicrobial and cytotoxic activity and the tested materials presented equivalent properties to those conventionally used in dentistry.