Melatonin protects TEGDMA-induced preodontoblast mitochondrial apoptosis via the JNK/MAPK signaling pathway.

Resin monomer-induced dental pulp injury presents a pathology related to mitochondrial dysfunction. Melatonin has been regarded as a strong mitochondrial protective bioactive compound from the pineal gland. However, it remains unknown whether melatonin can prevent dental pulp from resin monomer-induced injury. The aim of this study is to investigate the effects of melatonin on apoptosis of mouse preodontoblast cells (mDPC6T) induced by triethylene glycol dimethacrylate (TEGDMA), a major component in dental resin, and to determine whether the JNK/MAPK signaling pathway mediates the protective effect of melatonin. A well-established TEGDMA-induced mDPC6T apoptosis model is adopted to investigate the preventive function of melatonin by detecting cell viability, apoptosis rate, expressions of apoptosis-related proteins, mitochondrial ROS (mtROS) production, mitochondrial membrane potential (MMP) and adenosine triphosphate (ATP) level. Inhibitors of MAPKs are used to explore which pathway is involved in TEGDMA-induced apoptosis. Finally, the role of the JNK/MAPK pathway is verified using JNK agonists and antagonists. Our results show that melatonin attenuates TEGDMA-induced mDPC6T apoptosis by reducing mtROS production and rescuing MMP and ATP levels. Furthermore, mitochondrial dysfunction and apoptosis are alleviated only by the JNK/MAPK inhibitor SP600125 but not by other MAPK inhibitors. Additionally, melatonin downregulates the expression of phosphorylated JNK and counteractes the activating effects of anisomycin on the JNK/MAPK pathway, mimicking the effects of SP600125. Our findings demonstrate that melatonin protects mDPC6T cells against TEGDMA-induced apoptosis partly through JNK/MAPK and the maintenance of mitochondrial function, offering a novel therapeutic strategy for the prevention of resin monomer-induced dental pulp injury.

Protective Effect of Rutin on Triethylene Glycol Dimethacrylate-Induced Toxicity through the Inhibition of Caspase Activation and Reactive Oxygen Species Generation in Macrophages.

Rutin, also called quercetin-3-rhamnosyl glucoside, is a natural flavonol glycoside present in many plants. Rutin is used to treat various diseases, such as inflammation, diabetes, and cancer. For polymeric biomaterials, triethylene glycol dimethacrylate (TEGDMA) is the most commonly used monomer and serves as a restorative resin, a dentin bonding agent and sealant, and a bone cement component. Overall, TEGDMA induces various toxic effects in macrophages, including cytotoxicity, apoptosis, and genotoxicity. The aim of this study was to investigate the protective mechanism of rutin in alleviating TEGDMA-induced toxicity in RAW264.7 macrophages. After treatment with rutin, we assessed the cell viability and apoptosis of TEGDMA-induced RAW264.7 macrophages using an methylthiazol tetrazolium (MTT) assay and Annexin V-FITC/propidium iodide assay, respectively. Subsequently, we assessed the level of genotoxicity using comet and micronucleus assays, assessed the cysteinyla aspartate specific proteinases (caspases) and antioxidant enzyme (AOE) activity using commercial kits, and evaluated the generation of reactive oxygen species (ROS) using a dichlorodihydrofluorescein diacetate (DCFH-DA) assay. We evaluated the expression of heme oxygenase (HO)-1, the expression of nuclear factor erythroid 2 related factor (Nrf-2), and phosphorylation of AMP activated protein kinase (AMPK) using the Western blot assay. The results indicated that rutin substantially reduced the level of cytotoxicity, apoptosis, and genotoxicity of TEGDMA-induced RAW264.7 macrophages. Rutin also blocked the activity of caspase-3, caspase-8, and caspase-9 in TEGDMA-stimulated RAW264.7 macrophages. In addition, it decreased TEGDMA-induced ROS generation and AOE deactivation in macrophages. Finally, we found that TEGDMA-inhibited slightly the HO-1 expression, Nrf-2 expression, and AMPK phosphorylation would be revered by rutin. In addition, the HO-1 expression, Nrf-2 expression, and AMPK phosphorylation was enhanced by rutin. These findings indicate that rutin suppresses TEGDMA-induced caspase-mediated toxic effects through ROS generation and antioxidative system deactivation through the Nrf-2/AMPK pathway. Therefore, rutin has the potential to serve as a novel antitoxicity agent for TEGDMA in RAW264.7 macrophages.

Evaluation of residual monomer release after polymerization of different restorative materials used in pediatric dentistry.

BACKGROUND: The choice of the restorative resin material to be used in pediatric dentistry is of a great importance due to the cytotoxic effects caused by residual monomers. In this study, it was aimed to investigate the amount of residual monomer released over time from different resin-based restorative materials, which are widely used in pediatric dentistry, by using high performance liquid chromatography with photodiode array detector (HPLC-PDA). METHODS: The compomers in all colors (Twinky Star and Glasiositte A2), two composites with different hybrid properties (Arabesk-GrandioSO), and RMGIC (Ionolux) samples with 2 × 5 mm diameters were prepared. The samples were polymerized with an LED light unit (CELALUX 2, VOCO, Cuxhaven, Germany) and then finishing-polishing procedures were applied. A total of 156 samples were obtained, 13 samples in each of the 12 groups. The amount of residual monomer (BIS-GMA; HEMA, TEGDMA, UDMA) (µg/mL) released into the 75% ethanol solution was determined at different times, (1st hour, 1st, 7th, 14th, and 21st day) by using HPLC-PDA. RESULTS: The residual monomer release continued on day 21 and BIS-GMA was the most released monomer in all groups. HEMA release showed a maximum increase in all the materials at day 7. The highest amount of residual monomer was detected in the gold-colored compomer. HEMA and BIS-GMA release from RMGIC was less than others in all time frames. CONCLUSIONS: The color and composition of resin-based restorative materials affect the amount of residual monomer. Pediatric dentists should prefer gold-colored compomers less than others as a restorative material, especially in deep cavities. More studies are needed about the subject.

Notoginsenoside R1 alleviates TEGDMA-induced mitochondrial apoptosis in preodontoblasts through activation of Akt/Nrf2 pathway-dependent mitophagy.

Incomplete polymerization or biodegradation of dental resin materials results in the release of resin monomers such as triethylene glycol dimethacrylate (TEGDMA), causing severe injury of dental pulp cells. To date, there has been no efficient treatment option for this complication, in part due to the lack of understanding of the mechanism underlying these phenomena. Here, for the first time, we found that notoginsenoside R1 (NR1), a bioactive ingredient extracted from Panax notoginseng, exerted an obvious protective effect on TEGDMA-induced mitochondrial apoptosis in the preodontoblast mDPC6T cell line. In terms of the mechanism of action, NR1 enhanced the level of phosphorylated Akt (protein kinase B), resulting in the activation of a transcriptional factor, nuclear factor erythroid 2-related factor 2 (Nrf2), and eventually upregulating cellular ability to resist TEGDMA-related toxicity. Inhibiting the Akt/Nrf2 pathway by pharmaceutical inhibitors significantly decreased NR1-mediated cellular antioxidant properties and aggravated mitochondrial oxidative damage in TEGDMA-treated cells. Interestingly, NR1 also promoted mitophagy, which was identified as the potential downstream of the Akt/Nrf2 pathway. Blocking the Akt/Nrf2 pathway inhibited mitophagy and abolished the protection of NR1 on cells exposed to TEGDMA. In conclusion, these findings reveal that the activation of Akt/Nrf2 pathway-mediated mitophagy by NR1 might be a promising approach for preventing resin monomer-induced dental pulp injury.

Influence of TEGDMA monomer on MMP-2, MMP-8, and MMP-9 production and collagenase activity in pulp cells.

OBJECTIVES: Resin-based composites may leach monomers such as triethylene-glycol dimethacrylate (TEGDMA), which could contribute to intrapulpal inflammation. The aim of this investigation was to examine whether various concentrations of TEGDMA are able to influence dentally relevant Matrix metalloproteinase (MMP)-2, MMP-8, and MMP-9 production, total collagenase/gelatinase activity in pulp cells, and suggest possible signaling mechanisms. MATERIALS AND METHODS: Pulp cells were cultured, followed by a 1-day exposure to sublethal TEGDMA concentrations (0.1, 0.2, and 0.75 mM). Total MMP activity was measured by an EnzCheck total collagenase/gelatinase assay, while the production of specific MMPs and the relative changes of phosphorylated, i.e., activated signaling protein levels of extracellular signal-regulated kinase (ERK)1/2, p38, c-Jun N-terminal kinase (JNK) were identified by western blot. Immunocytochemistry image data was also plotted and analyzed to see whether TEGDMA could possibly alter MMP production. RESULTS: An increase in activated MMP-2, MMP-8, and MMP-9 production as well as total collagenase activity was seen after a 24-h exposure to the abovementioned TEGDMA concentrations. Increase was most substantial at 0.1 (P = 0.002) and 0.2 mM (P = 0.0381). Concurrent p-ERK, p-p38, and p-JNK elevations were also detected. CONCLUSIONS: Results suggest that monomers such as TEGDMA, leached from resin-based restorative materials, activate and induce the production of dentally relevant MMPs in pulp cells. Activation of ERK1/2, p38, or JNK and MMP increase may play a role in and/or can be part of a broader stress response. Clinical relevance Induction of MMP production and activity may further be components in the mechanisms of intrapulpal monomer toxicity.

Illuminating the cellular and molecular mechanism of the potential toxicity of methacrylate monomers used in biomaterials.

The cytotoxicity of methacrylate-based biopolymers crosslinked by in situ photopolymerization has been attributed mainly to residual methacrylate monomers released due to incomplete polymerization. The residual monomers, primarily triethyleneglycol dimethacrylate or 2-hydroxyethyl methacrylate, may irritate adjacent tissue, or be released into the bloodstream and reach practically all tissues. Increased production of reactive oxygen species, which may be connected to concomitant glutathione depletion, has been the most noticeable effect observed in vitro following the exposure of cells to methacrylates. Radical scavengers such as glutathione or N-acetylcysteine represent the most important cellular strategy against methacrylate-induced toxicity by direct adduct formation, resulting in monomer detoxification. Reactive oxygen species may participate in methacrylate-induced genotoxic or pro-apoptotic effects and cell-cycle arrest via induction of corresponding molecular pathways in cells. A deeper understanding of the biological mechanisms and effects of methacrylates widely used in various bioapplications may enable a better estimation of potential risks and thus, selection of a more appropriate composition of polymer material to eliminate potentially harmful substances such as triethyleneglycol dimethacrylate.

Evaluation of Monomer Leaching from a Resin Cement Through Dentin.

The aim of the study was to evaluate the elution of Triethylene glycol dimethacrylate (TEGDMA), Urethane dimethacrylate (UDMA), Bisphenol A glycerolate dimethacrylate (BisGMA), and Bisphenol A (BPA), from a dual-cured resin cement through human dentin, under constant positive pulpal pressure. Ten human dentin disks were adjusted into a custom made testing device and transparent glass slabs were luted with Variolink II cement, under a steady pressure. The device was filled with Ringer's solution and a pressure of 14.1 cm H2O was applied. Eluates were retrieved from each one of the ten specimens at 9 time interval. All the samples were analyzed by High Performance Liquid Chromatography (HPLC). TEGDMA was detected from the second and UDMA was detected from the fourth time interval and then. The highest average concentration of TEGDMA and UDMA was detected in the 3 day time interval. Time had a significant effect on their elution. BPA and BisGMA were not detected in any sample of any time interval. The clinical relevance of the present study is that the concentration of the eluted monomers, under the conditions that were chosen, did not reach toxic levels for the pulp.

The dental resin monomers HEMA and TEGDMA have inhibitory effects on osteoclast differentiation with low cytotoxicity.

The dental resin monomers 2-hydroxyethyl methacrylate (HEMA) and triethylene glycol dimethacrylate (TEGDMA) are released from the resin matrix due to unpolymerized monomers; once released, they influence various biological functions and the viability of cells in the oral environment. Although HEMA and TEGDMA have various effects on cells, including inflammation, inhibition of cell proliferation or differentiation, and apoptosis, the effects of these monomers on osteoclasts remain unknown. In this study, we investigated the effects of HEMA and TEGDMA on osteoclast differentiation of bone marrow-derived macrophages or murine monocytic cell line RAW-D. Both HEMA and TEGDMA inhibited osteoclast formation and their bone-resorbing activity at non-cytotoxic concentrations. Moreover, HEMA and TEGDMA decreased the expression of nuclear factor of activated T cells cytoplasmic-1 (NFATc1), a master regulator of osteoclast differentiation, and of osteoclast markers that are transcriptionally regulated by NFATc1, including Src and cathepsin K. Regarding their effects on signaling pathways involved in osteoclast differentiation, HEMA impaired the phosphorylation of extracellular signal-regulated kinase and Jun N-terminal kinase, whereas TEGDMA attenuated the phosphorylation of Akt and Jun N-terminal kinase. Thus, HEMA and TEGDMA inhibit osteoclast differentiation through different signaling pathways. This is the first report on the effects of the monomers HEMA and TEGDMA on osteoclasts. Copyright © 2017 John Wiley & Sons, Ltd.

Cytotoxicity and genotoxicity of triethyleneglycol-dimethacrylate in macrophages involved in DNA damage and caspases activation.

Triethyleneglycol-dimethacrylate (TEGDMA) is a monomer and widely used in dental composite resins. TEGDMA has been found to exhibit cytotoxicity and genotoxicity on many cells. However, little is known about the potential toxicological implications of TEGDMA on murine macrophage cell line RAW264.7. In this study, TEGDMA demonstrated a cytotoxic effect to RAW264.7 cells in a concentration- and time-dependent manner (p < 0.05). TEGDMA was found to induce two modes of cell death in a concentration-dependent manner (p < 0.05). TEGDMA-induced cell apoptosis was demonstrated by the increase in the portion of sub-G0/G1 phase and DNA ladder formation. In addition, TEGDMA exhibited genotoxicity via a dose-related increase in the numbers of micronucleus and DNA strand breaks (p < 0.05). Furthermore, the activation of caspase-3, -8, and -9 were generated by TEGDMA in a dose-dependent manner (p < 0.05). These results indicated that cytotoxicity and genotoxicity induced by TEGDMA in macrophages may be via DNA damage and caspase activation.

Reproductive toxicity evaluation of the dental resin monomer triethylene glycol dimethacrylate (CASRN 109-16-0) in mice.

The reproductive toxicity potential of the resin monomer triethylene glycol dimethacrylate (TEGDMA; Chemical Abstracts Service Registry Number 109-16-0) was investigated in male and female Crl:CD1(ICR) mice, 4 dosage groups, 25 mice/sex/group. Formulations of TEGDMA (0, 0.01, 0.1, or 1.0 mg/kg/d) in reverse osmosis-processed deionized water were intubated once daily beginning 28 days before cohabitation and continuing through mating (males) or through gestation day 17 (females). The following parameters were evaluated: viability, clinical signs, body weights, estrous cyclicity, necropsy observations, organ weights, sperm concentration/motility/morphology, cesarean-sectioning and litter observations, and histopathological evaluation of select tissues. No deaths or clinical signs related to TEGDMA occurred. No significant changes in male and female body weights and body weight gains were recorded for any of the administered dosages of TEGMDA. All mating and fertility parameters and all litter and fetal data were considered to be unaffected by dosages of TEGMDA as high as 1 mg/kg/d. Gross or histopathologic tissue changes attributable to the test article were not observed. Reproductive and developmental no observed adverse effect levels (NOAELs) for TEGMDA were 1.0 mg/kg/d, the highest dose tested. Comparison of conservatively estimated TEGDMA exposures from dental treatments to the NOAEL of 1.0 mg/kg/d identified in this study indicates margins of exposure of at least 120- to 3000-fold depending on the exposure scenario. The results of this study support the continued safe use of TEGDMA in polymeric dental products applied according to the manufacturers' instructions.

Investigation of Mechanical Properties of Polymer-Infiltrated Tetrapodal Zinc Oxide in Different Variants.

The aim of this study was to evaluate the influence of weight ratio, the shape of the precursor particles, and the application of a phosphate-monomer-containing primer on the mechanical properties of polymer infiltrated ceramic networks (PICNs) using zinc oxide. Two different types of zinc oxide particles were used as precursors to produce zinc oxide networks by sintering, each with two different densities resulting in two different weight ratios of the PICNs. For each of these different networks, two subgroups were built: one involving the application of a phosphate-monomer-containing primer prior to the infiltration of Bis-GMA/TEGDMA and one without. Elastic modulus and flexural strength were determined by using the three-point bending test. Vertical substance loss determined by the chewing simulation was evaluated with a laser scanning microscope. There was a statistically significant influence of the type of precursor particles on the flexural strength and in some cases on the elastic modulus. The application of a primer lead to a significant increase in the flexural strength and in most cases also in the elastic modulus. A higher weight ratio of zinc oxide led to a significantly higher elastic modulus. Few statistically significant differences were found for the vertical substance loss. By varying the shape of the particles and the weight fraction of zinc oxide, the mechanical properties of the investigated PICN can be controlled. The use of a phosphate-monomer-containing primer strengthens the bond between the infiltrated polymer and the zinc oxide, thus increasing the strength of the composite.

Advancing Dimethacrylate Dental Composites by Synergy of Pre-Polymerized TEGDMA Co-Filler: A Physio-Mechanical Evaluation.

Dental resin composites (DRCs) have gained immense popularity as filling material in direct dental restorations. They are highly valued for their ability to closely resemble natural teeth and withstand harsh oral conditions. To increase the clinical performance of dental restorations, various fillers are incorporated into DRCs. Herein, the effect of incorporating pre-polymerized triethylene glycol dimethacrylate (P-TEGDMA) as a co-filler in varying proportions (0%, 2.5%, 5%, and 10% by weight) into bisphenol A-glycidyl methacrylate (BisGMA)/TEGDMA/SiO2 resin composite was investigated. The obtained DRCs were examined for morphology, rheological properties, degree of crosslinking (DC), Vickers microhardness (VMH), thermal stability, and flexural strength (FS). The results revealed that SiO2 and P-TEGDMA particles were uniformly dispersed. The introduction of P-TEGDMA particles (2.5 wt.%) into the resin composite had a remarkable effect, leading to a significant reduction (p ≤ 0.05) in complex viscosity, decreasing from 393.84 ± 21.65 Pa.s to 152.84 ± 23.94 Pa.s. As a result, the DC was significantly (p ≤ 0.05) improved from 61.76 ± 3.80% to 68.77 ± 2.31%. In addition, the composite mixture demonstrated a higher storage modulus (G') than loss modulus (G″), indicative of its predominantly elastic nature. Moreover, the thermal stability of the DRCs was improved with the addition of P-TEGDMA particles by increasing the degradation temperature from 410 °C to 440 °C. However, the VMH was negatively affected. The study suggests that P-TEGDMA particles have the potential to be used as co-fillers alongside other inorganic fillers, offering a means to fine-tune the properties of DRCs and optimize their clinical performance.

Fabrication of Novel Pre-Polymerized BisGMA/Silica Nanocomposites: Physio-Mechanical Considerations.

Resin composite mimics tooth tissues both in structure and properties, and thus, they can withstand high biting force and the harsh environmental conditions of the mouth. Various inorganic nano- and micro-fillers are commonly used to enhance these composites' properties. In this study, we adopted a novel approach by using pre-polymerized bisphenol A-glycidyl methacrylate (BisGMA) ground particles (XL-BisGMA) as fillers in a BisGMA/triethylene glycol dimethacrylate (TEGDMA) resin system in combination with SiO2 nanoparticles. The BisGMA/TEGDMA/SiO2 mixture was filled with various concentrations of XL-BisGMA (0, 2.5, 5, and 10 wt.%). The XL-BisGMA added composites were evaluated for viscosity, degree of conversion (DC), microhardness, and thermal properties. The results demonstrated that the addition of a lower concentration of XL-BisGMA particles (2.5 wt.%) significantly reduced (p ≤ 0.05) the complex viscosity from 374.6 (Pa·s) to 170.84. (Pa·s). Similarly, DC was also increased significantly (p ≤ 0.05) by the addition of 2.5 wt.% XL-BisGMA, with the pristine composite showing a DC of (62.19 ± 3.2%) increased to (69.10 ± 3.4%). Moreover, the decomposition temperature has been increased from 410 °C for the pristine composite (BT-SB0) to 450 °C for the composite with 10 wt.% of XL-BisGMA (BT-SB10). The microhardness has also been significantly reduced (p ≤ 0.05) from 47.44 HV for the pristine composite (BT-SB0) to 29.91 HV for the composite with 2.5 wt.% of XL-BisGMA (BT-SB2.5). These results suggest that a XL-BisGMA could be used to a certain percentage as a promising filler in combination with inorganic fillers to enhance the DC and flow properties of the corresponding resin-based dental composites.

Investigation of antimicrobial and mechanical effects of functional nanoparticles in novel dental resin composites.

OBJECTIVES: Imidazole and benzimidazole derivatives have recently attracted attention as remarkable materials due to their advantages in chemistry, pharmacology, and biomaterials. This article focuses on dental composites with azole functional groups incorporated to affect their physicochemical and mechanical properties and antibacterial activity. METHODS: Dental composites were fabricated by embedding the functionalized imidazole and benzimidazole nanoparticles into a Bis-GMA/TEGDMA matrix to form the imidazole and benzimidazole dental composites series (I and B). The material was produced through hand blending of the monomer (50:50, wt%), filler (0-30, wt%), and initiator combination (CQ/EDMAB:0.8:1.6, wt%), and LED light-curing unit for 60 s. RESULTS: Using various characterization techniques, I and B series were validated. The dental composites' approximate solubility and sorption significances were evaluated by conducting experiments on specific dental composite formulations. Fenton reaction test was performed to determine the chemical stability of the dental composites. The mechanical properties of the dental composites were investigated. Finally, by testing cell growth in the presence of composites, their antibacterial activities were determined. CONCLUSIONS: In this study, it was observed that the mechanical, physiochemical, and antibacterial properties of the functional azole-containing nanoparticles were positively improved by adding them to the structure of dental composites. These experimental results paved the way for the synthesized materials to be used in industrial applications. CLINICAL SIGNIFICANCE: Since the chemical, mechanical, and antimicrobial properties of dental composites containing 10% imidazole and benzimidazole functional nanoparticles are far superior, they constitute an excellent alternative for preventing dental caries and long-term use of dental composites.

An Insight into Enamel Resin Infiltrants with Experimental Compositions.

Resin infiltration is a conservative treatment of initial enamel carious lesions. Only one infiltrant material is available on the market (Icon, DMG), and research is now investigating new chemical compositions so as to further exploit the benefits of the resin infiltration technique. A literature search of the articles testing the effects of different formulations on mechanical properties, resin penetration ability, remineralizing, and antibacterial activities was conducted. Of 238 articles, 29 resulted in being eligible for the literature review. The formulations investigated were all different and consisted in the inclusion of hydrophobic monomers (i.e., BisEMA, UDMA), solvents (ethanol, HEMA), alternative etchants (PAM) or molecules with antibacterial or bioactivity features (i.e., AgNP, YbF3, MTZ, chitosan, DMAMM, HAp, MC-IL, NACP, PUA, CHX) and microfilled resins. Information on the long-term performances of the tested experimental materials were scarce. The combination of TEGDMA with hydrophobic monomers and the inclusion of a solvent alternative to ethanol reinforced mechanical properties of the materials. Hybrid-glass materials demonstrated an enhanced remineralization capacity. Techniques such as tunnelization increased the penetration depth and preserved the recourse to less-conservative treatments. Combining the min-invasive infiltrant approach with remineralizing and bacteriostatic properties would be beneficial for therapeutic and economical aspects, according to the principles of minimally invasive dentistry.

The Influence of Low-Molecular-Weight Monomers (TEGDMA, HDDMA, HEMA) on the Properties of Selected Matrices and Composites Based on Bis-GMA and UDMA.

Bisphenol A-glycidyl methacrylate (bis-GMA) and urethane dimethacrylate (UDMA) are usually combined with low-viscosity monomers to obtain more desirable viscosity, handling characteristics and general properties. The present study determined the flexural strength (FS), flexural modulus (FM), diametral tensile strength (DTS), and hardness (HV) of five matrices and composites based on these resins. The polymerization shrinkage stress (PSS) was also studied for the composites. The polymer matrices were formed using bis-GMA and UDMA. TEGDMA, HEMA and HDDMA acted as co-monomers. The composites had 45 wt.% of filler content. The highest FS and FM were obtained from the UDMA/bis-GMA/TEGDMA/HEMA matrix and the composite (matrix + filler). The best DTS values were obtained from the UDMA/bis-GMA/HEMA matrix and the composite. One of the lowest values of FS, FM, and DTS was obtained from the UDMA/bis-GMA/HDDMA matrix and the composite. All the composites demonstrated similar hardness values. The lowest polymerization shrinkage stress was observed for the UDMA/bis-GMA/TEGDMA/HEMA composite, and the highest PSS was observed for the UDMA/bis-GMA/TEGDMA/HDDMA composite. The addition of HEMA had a positive effect on the properties of the tested materials, which may be related to the improved mobility of the bis-GMA and UDMA monomers.

TEGDMA (Triethylene Glycol Dimethacrylate) Induces Both Caspase-Dependent and Caspase-Independent Apoptotic Pathways in Pulp Cells.

Monomers leached from resin-based composites (RBCs) may reach intrapulpal concentrations of the millimolar (mM) range, which could contribute to inflammation. The aim of this investigation was to assess the cytotoxicity of triethylene glycol dimethacrylate (TEGDMA) monomers on pulp cells as well as to identify molecular mechanisms leading to apoptosis. Pulp cells were harvested from molars extracted for orthodontic reasons and cultured through an explant method. To assess cytotoxicity, cells underwent a 5-day exposure to 0.75, 1.5, and 3 mM TEGDMA and were subject to cell counting and WST-1 staining. Based on the findings, cells were subsequently exposed to 0.1, 0.2, 0.75, 1.5, and 3 mM TEGDMA for 24 h to uncover the details of apoptosis. Changes in the production or cleavage of the apoptosis-specific proteins caspase-8, caspase-9, caspase-3, caspase-12, and Apoptosis-Inducing Factor (AIF) were measured by Western blot. The 5-day study showed concentration- and time-dependent cytotoxicity. Significant cell death was detected after 24 h with TEGDMA concentrations of 1.5 and 3 mM. One-day exposure to TEGDMA led to the activation of caspase-8, -9, -3, and -12 and an increased AIF production. Results suggest that relevant concentrations of TEGDMA monomers, leached from RBCs, induce apoptosis in pulp cells through both caspase-dependent as well as caspase-independent mechanisms. Endoplasmic reticulum stress and the activation of caspase-independent apoptotic pathways may be further mechanisms by which monomers induce apoptosis in pulp cells.

Cytotoxic and Genotoxic Effects of Composite Resins on Cultured Human Gingival Fibroblasts.

The aim of the study was to evaluate the cytotoxic and genotoxic potential of five commercially available dental composite resins (CRs), investigating the effect of their quantifiable bisphenol-A-glycidyl-methacrylate (Bis-GMA) and/or triethylene glycol dimethacrylate (TEGDMA) release. Experiments were performed using the method of soaking extracts, which were derived from the immersion of the following CRs in the culture medium: Clearfil-Majesty-ES-2, GrandioSO, and Enamel-plus-HRi (Bis-GMA-based); Enamel-BioFunction and VenusDiamond (Bis-GMA-free). Human Gingival Fibroblasts (hGDFs) were employed as the cellular model to mimic in vitro the oral cavity milieu, where CRs simultaneously release various components. Cell metabolic activity, oxidative stress, and genotoxicity were used as cellular outcomes. Results showed that only VenusDiamond and Enamel-plus-HRi significantly affected the hGDF cell metabolic activity. In accordance with this, although no CR-derived extract induced a significantly detectable oxidative stress, only VenusDiamond and Enamel-plus-HRi induced significant genotoxicity. Our findings showed, for the CRs employed, a cytotoxic and genotoxic potential that did not seem to depend only on the actual Bis-GMA or TEGDMA content. Enamel-BioFunction appeared optimal in terms of cytotoxicity, and similar findings were observed for Clearfil-Majesty-ES-2 despite their different Bis-GMA/TEGDMA release patterns. This suggested that simply excluding one specific monomer from the CR formulation might not steadily turn out as a successful approach for improving their biocompatibility.

JNK-mediated blockage of autophagic flux exacerbates the triethylene glycol dimethacrylate-induced mitochondrial oxidative damage and apoptosis in preodontoblast.

Mitochondrial dependent oxidative stress (OS) and subsequent cell death are considered as the major cytotoxicity caused by Triethylene glycol dimethacrylate (TEGDMA), a commonly monomer of many resin-based dental composites. Under OS microenvironment, autophagy serves as a cell homeostatic mechanism and maintains redox balance through degradation or turnover of cellular components in order to promote cell survival. However, whether autophagy is involved in the mitochondrial oxidative damage and apoptosis induced by TEGDMA, and the cellular signaling pathways underlying this process remain unclear. In the present study, we demonstrated that TEGDMA induced mouse preodontoblast cell line (mDPC6T) dysfunctional mitochondrial oxidative response. In further exploring the underlying mechanisms, we found that TEGDMA impaired autophagic flux, as evidenced by increased LC3-II expression and hindered p62 degradation, thereby causing both mitochondrial oxidative damage and cell apoptosis. These results were further verified by treatment with chloroquine (autophagy inhibitor) and rapamycin (autophagy promotor). More importantly, we found that the JNK/MAPK pathway was the key upstream regulator of above injury process. Collectively, our finding firstly demonstrated that TEGDMA induced JNK-dependent autophagy, thereby promoting mitochondrial dysfunction-associated oxidative damage and apoptosis in preodontoblast.

Response of pulp cells to resin infiltration of enamel white spot-like lesions.

OBJECTIVES: To investigate the trans-enamel and trans-dentinal biological effects of treating enamel white spot-like lesions (EWSLs) with resin infiltration components (RICs) on odontoblast-like cells (MDPC-23) and human dental pulp cells (HDPCs). METHODS: EWSLs were induced in 60 enamel/dentin discs (4.0 ± 0.2 mm thick) using S. mutans. The discs were adapted into artificial pulp chambers and MDPC-23 were seeded on the dentin surface. The components of a resin infiltration system (Icon) were applied individually or in combination on the enamel surface as following (n = 10/treatment): Etch, Infiltrant, Etch+Infiltrant, or Etch+Dry+Infiltrant. The application of water or hydrogen peroxide served as negative and positive controls, respectively. After 72 h, MDPC-23 viability was evaluated. The extracts were exposed for 72 h to pre-cultured MDPC-23 and HDPCs in 96-well plates to evaluate cell viability, alkaline phosphatase activity (ALP), mineralized nodule formation (MN), and the expression of inflammatory cytokines (ICs) and mineralization-related genes (MRs). Data were analyzed by ANOVA complemented with Tukey or Games-Howell post-hocs (α = 5%). RESULTS: Cell viability, ALP activity, and MN formation were significantly reduced in response to the RICs, presenting intermediate values compared to positive and negative controls. Likewise, ICs were upregulated, whereas MRs were downregulated. Among the RICs, the Etch component caused the most notorious detrimental effects. SIGNIFICANCE: Resin infiltration of EWSLs negatively affected the metabolism of pulp cells in vitro. Therefore, even though resin infiltration is a micro-invasive therapy for non-cavitated caries in enamel, it should be closely followed up seen that components may diffuse and unbalance pulp homeostasis.