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BACKGROUND: Calcium silicate-based bioceramics have been applied in endodontics as advantageous materials for years, many chemical components and new synthesizing methods were used to improve the base formulation of the materials for positively affecting the sealers properties. Recently, a novel biomaterial formulation, grounded in strontium silicate, has been introduced to the market, offering potential advancements in the field. OBJECTIVE: To comparatively analyze the cytotoxicity and cell migration effects of a novel strontium silicate-based bioceramic material (CRoot SP) and those of calcium silicate-based (iRoot SP) and epoxide amine resin (AH Plus) sealers on stem cells derived from rat apical papilla(rSCAPs). METHODS: rSCAPs were isolated and characterized in vitro and subsequently cultured in the presence of various concentrations of CRoot SP, iRoot SP and AH Plus extracts. Cytotoxicity was assessed by CCK-8 assay, and cell-migration capacity was assessed by using wound healing assays . RESULTS: No significant differences in cell viability were observed in the 0.02 mg/mL and 0.2 mg/mL sealer groups. The cell viability of CRoot SP was consistently greater than that of iRoot SP at concentrations of 5 mg/mL and 10 mg/mL across all time points. Maximum cytotoxic effect was noted on day 5 with 10 mg/mL AH Plus.The scratch was partly healed by cell migration in all groups at 24 h, and the 0.02 mg/mL, and 0.2 mg/mL CRoot SP exerted beneficial effects on rSCAPs migration. CONCLUSIONS: CRoot SP exhibited less cytotoxic than the iRoot SP and AH Plus extracts after setting. A lower concentration of CRoot SP thus promotes the cell migration capacity of rSCAPs, and it may achieve better tissue repair during root canal treatment.

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OBJECTIVES: The present study aimed to determine in silico toxicity predictions of test compounds from hydraulic calcium silicate-based sealers (HCSBS) and AH Plus and computationally simulate the interaction between these substances and mediators of periapical inflammation via molecular docking. MATERIALS AND METHODS: All chemical information of the test compounds was obtained from the PubChem site. Predictions for bioavailability and toxicity analyses were determined by the Molinspiration Cheminformatics, pkCSM, ProTox-II and OSIRIS Property Explorer platforms. Molecular docking was performed using the Autodock4 AMDock v.1.5.2 program to analyse interactions between proteins (IL-1ß, IL-6, IL-8, IL-10 and TNF-α) and ligands (calcium silicate hydrate, zirconium oxide, bisphenol-A epoxy resin, dibenzylamine, iron oxide and calcium tungstate) to establish the affinity and bonding mode between systems. RESULTS: Bisphenol-A epoxy resin had the lowest maximum dose tolerated in humans and was the test compound with the largest number of toxicological properties (hepatotoxicity, carcinogenicity and irritant). All systems had favourable molecular docking. However, the ligands bisphenol-A epoxy resin and dibenzylamine had the greatest affinity with the cytokines tested. CONCLUSION: In silico predictions and molecular docking pointed the higher toxicity and greater interaction with mediators of periapical inflammation of the main test compounds from AH Plus compared to those from HCSBS. CLINICAL RELEVANCE: This is the first in silico study involving endodontic materials and may serve as the basis for further research that can generate more data, producing knowledge on the interference of each chemical compound in the composition of different root canal sealers.

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The aim of this study was to evaluate cytotoxicity and genotoxicity of calcium-silicate based sealers and comparing them with a gold standard-an epoxy-based sealant. Two experimental cell lines were used, gingival fibroblasts (hGF) and monocyte/macrophage peripheral blood cell line (SC). The cytotoxicity (XTT assay) and genotoxicity (comet assay) were evaluated both after 24-h and 48-h incubation. Additionally, after 48-h incubation, the cell apoptosis and cell cycle progression was detected. BioRoot Flow induced a significant decrease in hGF cells viability compared to the negative control groups both after 24-h (p < 0.001) and 48-h incubation (p < 0.01). In group with SC cells, after 24-h incubation significant increase in cells viability was detected for AH Plus Bioceramic Sealer in comparison to negative control (p < 0.05). BioRoot Flow and BioRoot RCS can be considered potentially genotoxic for the hGF cells after 48-h incubation (> 20% DNA damage). BioRoot Flow and BioRoot RCS, may have potential genotoxic effects and induce apoptosis in hGF cells which may irritate periapical tissues, resulting in a delayed healing. The findings of the study would be useful in selection of an appropriate sealant for root canal filling without causing cytotoxicity and genotoxicity.

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The purpose of this study was to assess the effect of a calcium silicate-based sealers (CeraSeal) and an epoxy resin-based sealer (AH Plus) on cytotoxicity and cell migration of stem cell from the human apical papilla (hSCAPs) by using the Alamar Blue, Annexin V-FICT and wound healing assays. In Alamar Blue assay, hSCAPs exposed to undiluted CeraSeal extract had significantly higher cell viability compared with that observed when cells were treated with AH Plus in all experimental period (p < 0.001). The flow cytometry analysis confirmed the comparison on viable cells and indicated that AH Plus increased apoptosis compared to CeraSeal and the control groups (p < 0.001). Additionally, AH Plus exhibited significantly lower level of cell migration than CeraSeal and the control for up to 48 h observation (p < 0.01). In summary, calcium silicate-based sealer (CeraSeal) is less cytotoxic and more biocompatible than epoxy resin-based sealer (AH Plus).

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OBJECTIVE: The aim of this study was to investigate solubility, pH value, chemical structure, radiopacity, and cytotoxicity of AH Plus BC, TotalFill BC, AH Plus, and AH Plus Jet sealers. MATERIALS AND METHODS: Cytotoxicity analysis with direct and extraction tests at 3 different concentrations (1:1, 1:2, 1:4 v/v%) and time (24 h, 48 h, and 72 h) on Saos-2, PdLF, and THP-1 cell lines, chemical structure with scanning electron microscopy (SEM) and energy-dispersive X-ray spectroscopy (EDX) analysis, solubility, pH, and radiopacity values of AH Plus BC, TotalFill BC, AH Plus, and AH Plus Jet were evaluated. For statistical analyses of the groups, repeated measures, factorial, and one-way ANOVA tests were used. The statistical significance level was set at p < .05. RESULTS: Resin-based sealers showed higher cytotoxicity values than the bioceramic-based sealers (p < 0.05). Time and concentrations were effective on the cell viabilities for cell lines. Higher peaks of calcium were detected bioceramic-based sealers and higher amount of zirconium was detected in AH Plus BC (p < 0.05). AH Plus BC showed similar radiopacity value with AH Plus, AH Plus Jet, whereas TotalFill BC showed the lowest radiopacity (p < 0.05). Bioceramic-based sealers had higher pH values in all experiment periods, and the difference between resin- and bioceramic-based sealer groups was significant (p < 0.05). However, the solubility values of the tested root canal sealers revealed no differences (p > 0.05). CONCLUSIONS: The newly produced AH Plus BC Sealer showed similar properties with TotalFill BC, and their biological properties were better than AH Plus and AH Plus Jet. CLINICAL RELEVANCE: AH Plus BC could be a possible alternative to other bioceramic- or resin-based sealers.

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The function of root canal sealer was to achieve an appropriate three-dimensional filling effect by filling the root canal and some irregular lumen, thereby inhibiting the residual bacteria. There were many types of sealers, but research to find the most suitable ones was still ongoing. In recent years, researchers had continuously improved the performance of sealers by developing new sealers or adding active ingredients to the sealers. However, most sealers exhibit varying degrees of cytotoxicity and tissue responses, which affect clinical therapy efficacy. This review describes different technical approaches, and recent research progress in the biocompatibility evaluation of root canal sealers and provides brief insights into this field by summarising the performance studies of different root canal sealers.

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Periapical tissue may be exposed to root canal filling materials in consequence of root canal therapy. There is scant scientific data about the biocompatibility of root canal filling materials of various chemistry on the periapical area. This study aimed to investigate the effects of different root canal sealers and their eluates on human alveolar osteoblasts in terms of cell proliferation, adhesion, morphology and gene expression in vitro. Five endodontic sealers (AH Plus®, Apexit®, Tubli-Seal®, Real Seal SE®, EndoRez®) and one gutta-percha obturation material (BeeFill®) were tested. Human alveolar osteoblasts derived from 3 different donors following incubation with sealer eluates after 24 h and 72 h were investigated by means of qPCR (gene expression). Morphological reactions of the alveolar osteoblasts were measured by culturing the cells for 3 d, and 7 d and 14 d, respectively, followed by scanning electron microscopy (morphology, adhesion) and fluorescence imaging of the actin cytoskeleton (morphology, proliferation). A repeated measures analysis was performed and p-values were adjusted by Tukey. While all sealers influenced the cell morphology and the expression of genes associated with apoptosis (Casp3), proliferation (histone H3), and inflammation (interleukin-6 and matrix metalloproteinases 1 and 3), mainly AH Plus® and Apexit® yielded a regular actin cytoskeleton and beneficial gene expression patterns. Regarding cell adhesion, only AH Plus® supported proper anchorage for alveolar osteoblasts. Our results provide evidence for the biocompatibility of epoxy resin-based endodontic sealers, i.e. AH Plus®, while other sealers proved cytotoxic for alveolar osteoblasts. Further studies are needed for understanding the bone cell reactions after endodontic treatment and the clinical decision-making regarding the sealer of choice for root canal fillings.

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We describe a novel nature-derived epoxy resin monomer (ERM) derived from the plant lignan pinoresinol. Epoxy resins are thermosetting materials in global usage owing to their excellent technical properties such as flexibility and durability. However, their adverse health effects are often not considered and affect users of epoxy resins worldwide. Components of epoxy resin systems are strong skin sensitizers and cause allergic contact dermatitis. The reported prevalence attributable to epoxy chemicals is between 11.7 and 12.5% of all cases of occupational allergic contact dermatitis. We are committed to developing epoxy resins with reduced allergenic effect, while maintaining their excellent properties. The novel ERM, pinoresinol diglycidyl ether (PinoDGE), was synthesized in one step from pinoresinol and epichlorohydrin in 88% yield. It was not classified as a skin sensitizer in the in vivo local lymph node assay, at concentrations up to 0.17 m, as it did not cause a stimulation index >3 compared to control. Pinoresinol diglycidyl ether reacted with the model peptide AcPHCKRM in a reactivity assay and was predicted to be a skin sensitizer in the KeratinoSens assay. Preliminary cross-linking studies indicate that it has promising properties compared to commercially used ERMs. Pinoresinol diglycidyl ether could be seen as a lead compound for further development of alternative ERMs with a better safety profile based on natural and renewable sources for construction of epoxy resin polymers.

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This study evaluated the cytotoxicity, the antimicrobial and physicochemical properties of root canal sealers incorporated with phytotherapic Uncaria tomentosa (UT). Unmodified AH Plus (Dentsply, DeTrey, Germany) and MTA Fillapex (Angelus, Londrina, Brazil) were used as controls. UT was incorporated into AH Plus and MTA Fillapex, at concentrations of 2% and 5% of the total weight of these sealers (w/w). Flowability, setting time, and solubility were evaluated following ISO requirements. The pH values were measured at periods of 12, 24, 48 hours, and 7 days. The antimicrobial activity of the sealers against Enterococcus faecalis was analyzed by both direct contact tests in freshly prepared sealers, and after 7 days. The cytotoxicity of the samples was evaluated by the MTT assay, to check Balb/c 3T3 cell viability. The statistical analysis was performed by one-way ANOVA and Tukey's test (p < 0.05). The incorporation of UT was associated with a decrease in flow, for both sealers, an increase in AH Plus setting time, increase in MTA Fillapex pH values, and solubility (after 14 days), for both sealers (p < 0.05). Regarding the antibacterial evaluation, bacterial reduction was reported after incorporation of UT into both AH Plus and MTA Fillapex, up to 7 days after handling of the material (P<0.05). UT incorporation decreased the cytotoxic effects of both AH Plus and MTA Fillapex sealers in a way directly proportional to their respective concentrations (p < 0.05). In conclusion, UT can be added to both sealers to reduce their cytotoxicity, and improve their antibacterial effects, without compromising their original physicochemical properties.

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Severe traumatic bleeding control and wound-related anti-infection play a crucial role in saving lives and promoting wound healing for both the military and the clinic. In this contribution, an inherent antibacterial and instant swelling ε-poly-lysine/poly (ethylene glycol) diglycidyl ether (EPPE) superabsorbent was developed by a simple mild ring-opening reaction. The as-prepared EPPE1 displayed a porous structure and rough surface and exhibited instant water-triggered expansion with approximately 6300% swelling ratio in deionized water. Moreover, EPPE1 presented efficient pro-coagulation capacity by hemadsorption that can facilitate blood cell gathering and activation in vitro and exhibited a shorter in vivo hemostasis time than that of commercial gelatin sponge and CELOX in both rat tail amputation and noncompressible rat liver lethal defect model. Also, EPPE1 showed excellent antibacterial capacity, prominent biocompatibility, and great biodegradability. Additionally, EPPE1 significantly promotes in vivo wound healing in a full-thickness skin defect model due to its great hemostasis behavior and remarkable bactericidal performance. Hence, EPPE has great potential for serving as an extensively applied hemostatic agent under varied clinical conditions.

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A composite wound dressing has been developed by combining different layers consisting of polymers and textiles. Wheat germ oil (WGO) loaded hydrogels have successfully formed on textile nonwovens by cross-linking sodium alginate (SA) with poly(ethylene glycol) diglycidyl ether (PEGDGE). Following freeze-drying, textile-hydrogel composites have been examined according to their physical properties, pH, fluid handling capacity, water vapour permeability, morphology, chemical structure, and cytotoxicity. Hydrogels containing WGO swelled less than pristine hydrogels. Samples with 1% WGO and no WGO showed swelling of 5.9 and 10.5 g/g after 8 h. WGO inclusion resulted in reduced, but more stable fluid handling properties, with more uniform pore distribution (100-200 µm). Moreover, the proliferation of NIH/3T3 cells significantly improved with 1% WGO contained hydrogels. Also, commercial self-adhesive dressings that secure the hydrogels to the wound area were investigated regarding transfer properties. The proposed product demonstrated 8.05 cm3/cm2/s and 541.37 g/m2/day air and water vapour permeability.

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Hyaluronic acid (HA) dermal fillers are produced by crosslinking HA with agents, such as 1,4-butanediol diglycidyl ether (BDDE) and poly (ethylene glycol) diglycidyl ether (PEGDE) to acquire desired properties. Thus, the safety evaluation of these crosslinkers is needed at the cellular level. In the present study, cell viability, cytotoxicity, membrane integrity, reactive oxygen species (ROS), mitochondrial membrane potential (MMP), and inflammatory responses were evaluated in the human keratinocyte cell line, HaCaT and human dermal fibroblast cell line, HDF in response to treatment with the crosslinkers. In both the cell lines, BDDE significantly decreased cell viability at 100-1000 ppm, while PEGDE showed a decrease at 500-1000 ppm. In HaCaT cells, BDDE markedly increased cytotoxicity (lactate dehydrogenase release) at 100-1000 ppm, but PEGDE showed an increase at 500-1000 ppm. Cells treated with BDDE (100 ppm) caused alteration in the integrity of cell membrane and shape. In both the cell lines, BDDE-treated cells showed significantly higher ROS levels and MMP loss than PEGDE-treated cells. Also, BDDE-treated cells exhibited higher COX-2 expression at 100 ppm. Expression of inflammatory cytokines (TNF-α, and IL-1 ß) was higher in BDDE-treated cells. Taken together, PEGDE-treated cells showed markedly lower cytotoxicity, ROS production, and inflammatory responses than BDDE-treated cells. Our data suggest that PEGDE is safer than BDDE as a crosslinker in HA dermal fillers.

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Abstract This study evaluated the cytotoxicity, the antimicrobial and physicochemical properties of root canal sealers incorporated with phytotherapic Uncaria tomentosa (UT). Unmodified AH Plus (Dentsply, DeTrey, Germany) and MTA Fillapex (Angelus, Londrina, Brazil) were used as controls. UT was incorporated into AH Plus and MTA Fillapex, at concentrations of 2% and 5% of the total weight of these sealers (w/w). Flowability, setting time, and solubility were evaluated following ISO requirements. The pH values were measured at periods of 12, 24, 48 hours, and 7 days. The antimicrobial activity of the sealers against Enterococcus faecalis was analyzed by both direct contact tests in freshly prepared sealers, and after 7 days. The cytotoxicity of the samples was evaluated by the MTT assay, to check Balb/c 3T3 cell viability. The statistical analysis was performed by one-way ANOVA and Tukey's test (p < 0.05). The incorporation of UT was associated with a decrease in flow, for both sealers, an increase in AH Plus setting time, increase in MTA Fillapex pH values, and solubility (after 14 days), for both sealers (p < 0.05). Regarding the antibacterial evaluation, bacterial reduction was reported after incorporation of UT into both AH Plus and MTA Fillapex, up to 7 days after handling of the material (P<0.05). UT incorporation decreased the cytotoxic effects of both AH Plus and MTA Fillapex sealers in a way directly proportional to their respective concentrations (p < 0.05). In conclusion, UT can be added to both sealers to reduce their cytotoxicity, and improve their antibacterial effects, without compromising their original physicochemical properties.

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The aim of the present study was to establish a standard methodology for the extraction of epoxy resin precursors from several types of food packages (cans, multi-layered composite material, and cups) with selected simulation media (distilled water, 5% ethanol, 3% dimethyl sulfoxide, 5% acetic acid, artificial saliva) at different extraction times and temperatures (factors). Biological analyses were conducted to determine the acute toxicity levels of the extracts (with Vibrio fischeri bacteria) and their endocrine potential (with Saccharomyces cerevisiae yeasts). In parallel, liquid chromatography-tandem mass spectrometry was performed to determine levels of bisphenol A diglycidyl ether (BADGE), bisphenol F diglycidyl ether (mixture of isomers, BFDGE), ring novolac glycidyl ether (3-ring NOGE), and their derivatives. The variation induced by the different experimental factors was statistically evaluated with analysis of variance simultaneous component analysis (ASCA). Our findings demonstrate the value of using a holistic approach to best partition the effects contributing to the end points of these assessments, and offer further guidance for adopting such a methodology, thus being a broadly useful reference for understanding the phenomena related to the impacts of food packaging materials on quality for long- and short-term storage, while offering a general method for analysis.

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The rapidly expanding fields of bioelectronics, and biological interfaces with electronic sensors and stimulators, are placing an increasing demand on candidate materials to serve as robust surfaces that are both biocompatible, stable and electroconductive. Amongst conductive polymers, poly(3,4-ethylenedioxythiophene):poly(styrenesulfonate) (PEDOT:PSS) is a promising material in biomedical research due to its appropriate stability and high conductivity, however its intrinsic solubility requires a crosslinking process that can limit its conductivity and biocompatibility. Poly(ethylene glycol) is known to be a suitably anti-immunogenic moiety and its derivatives have been widely used for biomedical applications. In this study we investigate the application of poly(ethylene glycol)diglycidyl ether (PEGDE) as an effective crosslinker and conductive filler for PEDOT:PSS. From our interpretation of XPS analysis we hypothesise that the crosslinking reaction is occurring via the epoxy ring of PEGDE interacting with the sulfonic groups of excel PSS chains, which reaches a saturation at 3 w/v% PEGDE concentration. PEGDE crosslinked films did not disperse in aqueous environments, had enhanced electrical conductivity and imparted a significant degree of hydrophilicity to PEDOT:PSS films. This hydrophilicity and the presence of biocompatible PEGDE led to good cell viability and a significantly increased degree of cell spreading on PEDOT:PSS films. In comparison to widely reported chemical crosslinking via glycidoxy propyltrimethoxysilane (GOPS), this original crosslinking yields a highly hydrophilic 2D film substrate with increased electroconductive and biocompatibility properties, resulting in a next-generation formulation for bioengineering applications.

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OBJECTIVES: The aim of this study was to evaluate the subcutaneous response induced by Roeko Guttaflow2 (RG), Sealapex Xpress (SX), AH Plus (AHP) sealers. METHODS: 100 BALB/c mice received implants in the subcutaneous tissue with the tested materials (10 animals per period for each evaluated sealer) and were evaluated after different experimental periods (7, 21 and 63 days), in each animal was placed a tube, the control group was an empty tube. Histological analysis evaluated semi-quantitatively the inflammatory infiltration, collagen fiber formation and tissue thickness. In addition, immunohistochemistry was performed for interleukin-6 (IL-6). Data were statistically analyzed (α = 0.05). RESULTS: RG promoted a greater collagen fiber formation at 7 days and 63 days compared to the CG (p = 0.004) and AHP (p = 0.005) respectively, while at 21 days, the SX promoted a greater reaction (p = 0.021). For the tissue thickness, there was a greater reaction at 7 days with CG (p = 0.0156) and with RG at 63 days (p = 0.03). Regarding the inflammatory infiltrate, there was no difference at 7 days and 63 days (p = 0.5; p = 0.27), while at 21 days, a statistically difference was found between SX, CG (p = 0.04) and RG (p = 0.027). In addition, the presence of IL-6 was observed in almost all groups, with a more intense marking at 7days. SIGNIFICANCE: All cements evaluated presented a satisfactory tissue response, however, RG was the one that presented a more satisfactory tissue response.

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The aim of the study was to evaluate the cytotoxicity of three epoxy resin-based endodontic sealer, AH Plus, Sicura Seal and Top Seal. Direct and indirect cytotoxicity were evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay and LIVE/DEAD® Viability/Cytotoxicity Assay on MG63 osteoblasts-like cells. Data were statistically analyzed by analysis of variance and Tukey test, setting a significance level of 5%. Both results related to direct and indirect cell viability tests showed that all groups were significantly more cytotoxic than the negative control group. The cytotoxicity activity after one week of culture showed the absence of direct cytotoxicity, while a medium rate of indirect cytotoxicity. All the three epoxy resin-based sealers (AH Plus, Top Seal and Sicura Seal) showed a medium rate of cytotoxicity on osteoblasts-like cells in vitro. No significant difference was found among the sealers analyzed.

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Bisphenol A (BPA) is used in the manufacture of polycarbonate and epoxy resin plastics. There has been interest in the possibility that BPA has immunotoxic properties, and a variety of investigations have explored this. Among the approaches taken have been human observational and cross-sectional studies, investigations using experimental animals, and in vitro studies, some of which have been reviewed previously by the European Food Safety Authority (EFSA). This commentary aims to provide a focused review of data regarding the ability of BPA to perturb the immune system, including the developing immune system, and to cause related adverse health effects. The objective is to complement the evaluations conducted by EFSA, with a focus on the ability of BPA to impair immune function, to promote respiratory allergy and airway inflammation, and to compromise immunological tolerance to dietary proteins. The conclusion drawn is that there is currently no persuasive evidence that BPA has significant immunotoxic potential. This is, in part, due to some of the data reviewed being apparently contradictory or inconsistent, and the investigations from which those data were derived having limitations with regard to experimental design. The conclusion drawn here is that presently there is no clear evidence that BPA has the potential to cause immunotoxicity resulting in adverse health effects.

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The environmental release of nanoparticles is attracting increasing attention. Graphene oxide (GO) embedded in epoxy resin (ER) is a popular composite that has been used in various fields, but the environmental release of GO-ER composites and the effects on organisms in the environment remain unknown. The present work found that GO-ER composites in water for 2-7 days resulted in the release of 0.3-2.1% GO-ER at nanoscale (2-3 nm thickness and approximately 70-130 nm lateral length). Interestingly, pristine GO quenched 30-45% hydroxyl and 12% nitroxide free radicals, whereas this capacity was not observed for the released particles from GO-ER. At environmentally relevant concentrations (µg/L), released GO-ER particles, but not GO or ER matrix, promoted algal reproduction by 34% and chlorophyll biosynthesis by 65-127% at 96 h. Released GO-ER entered algal cells and induced a slight increase in reactive oxygen species but did not elicit notable cell structure damage. The upregulated amino acids and phenylalanine metabolism, and the downregulated fatty acid biosynthesis contributed to algal growth promoted by released GO-ER. Previous studies of pristine nanoparticles were unable to reflect the environmental effects of released nanoparticles into the environment, and our research on the exposure-toxicological continuum adds important contributions to this field.

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