Assessing the role of the graphene family nanomaterials (GFNs: Graphene, GO, rGO) in modifying the toxicity potential and environmental risk of flame retardant, tetrabromobisphenol-A (TBBPA) in the marine microalgae Chlorella sp.

In recent years, a growing concern has emerged regarding the environmental implications of flame retardants (FRs) like tetrabromobisphenol-A (TBBPA) and graphene family nanomaterials (GFNs), such as graphene, graphene oxide (GO), and reduced graphene oxide (rGO), on marine biota. Despite these substances' well-established individual toxicity profiles, there is a notable gap in understanding the physicochemical interactions within the binary mixtures and consequent changes in the toxicity potential. Therefore, our research focuses on elucidating the individual and combined toxicological impacts of TBBPA and GFNs on the marine alga Chlorella sp. Employing a suite of experimental methodologies, including Raman spectroscopy, contact angle measurements, electron microscopy, and chromatography, we examined the physicochemical interplay between the GFNs and TBBPA. The toxicity potentials of individual constituents and their binary combinations were assessed through growth inhibition assays, quantifying reactive oxygen species (ROS) generation and malondialdehyde (MDA) production, photosynthetic activity analyses, and various biochemical assays. The toxicity of TBBPA and graphene-based nanomaterials (GFNs) was examined individually and in combinations. Both pristine TBBPA and GFNs showed dose-dependent toxicity. While lower TBBPA concentrations exacerbated toxicity in binary mixtures, higher TBBPA levels reduced the toxic effects compared to pristine TBBPA treatments. The principal mechanism underlying toxicity was ROS generation, resulting in membrane damage and perturbation of photosynthetic parameters. Cluster heatmap and Pearson correlation were employed to assess correlations between the biological parameters. Finally, ecological risk assessment was undertaken to evaluate environmental impacts of the individual components and the mixture in the algae.

EPS-corona formation on graphene family nanomaterials (GO, rGO and graphene) and its role in mitigating their toxic effects in the marine alga Chlorella sp.

GFNs have widespread applications but can harm marine systems due to excessive use and improper disposal. Algae-secreted EPS can mitigate nanomaterial harm, but their impact on GFN toxicity is understudied. Hence, in the present study, we investigated the toxicity of three GFNs, graphene oxide (GO), reduced graphene oxide (rGO), and graphene, in pristine and EPS-adsorbed forms in the marine alga Chlorella sp. At an environmentally relevant concentration of 1 mgL-1, all three GFNs induced considerable oxidative stress and impeded growth and photosynthetic activity of the algae. The order of the toxic potential followed GO > rGO > graphene. The various facets of adsorption of EPS (1:1 mixture of loosely bound, and tightly bound EPS) on GFNs were investigated through microscopy, surface chemical analyses, fluorescence quenching studies, and isotherm and kinetics studies. Amongst the pristine GFNs treated with algal cells, GO was found to exert the maximum negative effects on algal growth. Upon adsorption of EPS over the GFNs, a significant decline in growth inhibition was observed compared to the respective pristine forms which strongly correlated with reduced oxidative stress and enhanced photosynthetic parameters in the cells. The formation of a layer of eco-corona after interaction of GFNs with EPS possibly caused a barrier effect which in turn diminished their toxic potential. The findings from the present investigation offer valuable insights into the environmental toxicity of GFNs and show that the eco-corona formation may lessen the risk posed by these materials in the marine environment.

Fate and effects of graphene oxide alone and with sorbed benzo(a)pyrene in mussels Mytilus galloprovincialis.

Graphene oxide (GO) has gained a great scientific and economic interest due to its unique properties. As incorporation of GO in consumer products is rising, it is expected that GO will end up in oceans. Due to its high surface to volume ratio, GO can adsorb persistent organic pollutants (POPs), such as benzo(a)pyrene (BaP), and act as carrier of POPs, increasing their bioavailability to marine organisms. Thus, uptake and effects of GO in marine biota represent a major concern. This work aimed to assess the potential hazards of GO, alone or with sorbed BaP (GO+BaP), and BaP alone in marine mussels after 7 days of exposure. GO was detected through Raman spectroscopy in the lumen of the digestive tract and in feces of mussels exposed to GO and GO+BaP while BaP was bioaccumulated in mussels exposed to GO+BaP, but especially in those exposed to BaP. Overall, GO acted as a carrier of BaP to mussels but GO appeared to protect mussels towards BaP accumulation. Some effects observed in mussels exposed to GO+BaP were due to BaP carried onto GO nanoplatelets. Enhanced toxicity of GO+BaP with respect to GO and/or BaP or to controls were identified for other biological responses, demonstrating the complexity of interactions between GO and BaP.

Potential Biomedical Limitations of Graphene Nanomaterials.

Graphene-family nanomaterials (GFNs) possess mechanical stiffness, optical properties, and biocompatibility making them promising materials for biomedical applications. However, to realize the potential of graphene in biomedicine, it must overcome several challenges that arise when it enters the body's circulatory system. Current research focuses on the development of tumor-targeting devices using graphene, but GFNs accumulated in different tissues and cells through different pathways, which can cause toxic reactions leading to cell apoptosis and body dysfunction when the accumulated amount exceeds a certain limit. In addition, as a foreign substance, graphene can induce complex inflammatory reactions with immune cells and inflammatory factors, potentially enhancing or impairing the body's immune function. This review discusses the biomedical applications of graphene, the effects of graphene materials on human immune function, and the biotoxicity of graphene materials.

Development of Thiol-Maleimide hydrogels incorporating graphene-based nanomaterials for cancer chemo-photothermal therapy.

Nano-sized materials have been widely explored in the biomedicine field, especially due to their ability to encapsulate drugs intended to be delivered to cancer cells. However, systemically administered nanomaterials face several barriers that can hinder their tumor-homing capacity. In this way, researchers are now focusing their efforts in developing technologies that can deliver the nanoparticles directly into the tumor tissue. Particularly, hydrogels assembled using Thiol-Maleimide Michael type additions are emerging for this purpose due to their capacity to incorporate high nanoparticles' doses in a compact 3D structure as well as good chemical selectivity, biocompatibility, and straightforward preparation. Nevertheless, such hydrogels have been mostly prepared using synthetic polymers, which is not ideal due to their poor biodegradability. In this work, a novel natural polymer-based Thiol-Maleimide hydrogel was produced for application in breast cancer chemo-photothermal therapy. To obtain natural polymers compatible with this crosslinking chemistry, Hyaluronic acid was endowed with Thiol groups and deacetylated Chitosan was grafted with Maleimide groups. Parallelly, Doxorubicin loaded Dopamine-reduced graphene oxide (DOX/DOPA-rGO) was prepared for attaining Near Infrared (NIR) light responsive chemo-photothermal nanoagents. By simply mixing Hyaluronic Acid-Thiol, deacetylated Chitosan-Maleimide and DOX/DOPA-rGO, Thiol-Maleimide crosslinked hydrogels incorporating this nanomaterial could be assembled (DOX/DOPA-rGO@TMgel). When breast cancer cells were incubated with DOPA-rGO@TMgel and exposed to NIR light (photothermal therapy), their viability was reduced to about 59 %. On the other hand, DOX/DOPA-rGO@TMgel (chemotherapy) reduced cancer cells' viability to 50 %. In stark contrast, the combined action of DOX/DOPA-rGO@TMgel and NIR light decreased breast cancer cells' viability to just 21 %, highlighting its chemo-photothermal potential.

Graphene Family Nanomaterials for Stem Cell Neurogenic Differentiation and Peripheral Nerve Regeneration.

Stem cells play a critical role in peripheral nerve regeneration. Nerve scaffolds fabricated by specific materials can help induce the neurogenic differentiation of stem cells. Therefore, it is a potential strategy to enhance therapeutic efficiency. Graphene family nanomaterials are widely applied in repairing peripheral nerves. However, the mechanism underlying the pro-regeneration effects remains elusive. In this review, we first discuss the properties of graphene family nanomaterials, including monolayer and multilayer graphene, few-layer graphene, graphene oxide, reduced graphene oxide, and graphene quantum dots. We also introduce their applications in regulating stem cell differentiation. Then, we review the potential mechanisms of the neurogenic differentiation of stem cells facilitated by the materials. Finally, we discuss the existing challenges in this field to advance the development of nerve biomaterials.

The design, construction and application of graphene family composite nanocoating on dental metal surface.

Enhancement of the biological and mechanical properties of dental metals is important for accommodation with therapeutic schemes in different stomatological disciplines. Nanocoatings based on graphene family nanomaterials (GFNs) improve the topological structure and physicochemical properties of metal surfaces, endowing them with new properties while maintaining inherent mechanical properties. Nano-composite coatings, composed of GFNs with one or more type of polymer, metal, oxide, and inorganic nonmetallic compound, offer more matching modification schemes to meet multifunctional oral treatment requirements (e.g., anti-bacterial and anti-corrosive activity, osteogenesis and angiogenesis). This review describes recent progress in the development of GFN composite nanocoatings for the modification of dental metals, focus on biological effects in clinical settings. Underlying molecular mechanisms, critical modification schemes, and technical innovation in preparation methods are also discussed. The key parameters of GFN composite nanocoating surface modification are summarized according to effects on cellular responses and antibacterial activity. This review provides a theoretical reference for the optimization of the biological effects and application of GFN composite nanocoatings for dental metals, and the promotion of the environmentally friendly large-scale production of high-quality multifunctional GFN-based nanocoatings in the field of oral science.

The Effects of Graphene-Family Nanomaterials on Plant Growth: A Review.

Numerous reports of graphene-family nanomaterials (GFNs) promoting plant growth have opened up a wide range of promising potential applications in agroforestry. However, several toxicity studies have raised growing concerns about the biosafety of GFNs. Although these studies have provided clues about the role of GFNs from different perspectives (such as plant physiology, biochemistry, cytology, and molecular biology), the mechanisms by which GFNs affect plant growth remain poorly understood. In particular, a systematic collection of data regarding differentially expressed genes in response to GFN treatment has not been conducted. We summarize here the fate and biological effects of GFNs in plants. We propose that soil environments may be conducive to the positive effects of GFNs but may be detrimental to the absorption of GFNs. Alterations in plant physiology, biochemistry, cytological structure, and gene expression in response to GFN treatment are discussed. Coincidentally, many changes from the morphological to biochemical scales, which are caused by GFNs treatment, such as affecting root growth, disrupting cell membrane structure, and altering antioxidant systems and hormone concentrations, can all be mapped to gene expression level. This review provides a comprehensive understanding of the effects of GFNs on plant growth to promote their safe and efficient use.

Differential Toxicity of Graphene Family Nanomaterials Concerning Morphology.

Graphene family nanomaterials (GFNs) are well-known carbonaceous materials, which find application in several fields like optoelectronics, photocatalysis, nanomedicine, and tissue regeneration. Despite possessing many advantages in biomedical applications, GFNs exhibited toxicity depending on various parameters including dosage, size, exposure time, and kinds of administration. GFNS are majorly classified into nanosheets, quantum dots, nanoplatelets, and nanoribbons based on morphology. Understanding the toxic effects of GFNs would provide new suggestions as to how the materials can be utilized effectively. Hence, we are summarizing here some of the recent findings in cellular and animal level toxicity studies of GFNs on the perspective of their different morphologies. Notwithstanding, we highlight progress, challenges, and new toxicological approaches to ensure biosafety of GFNs for future directions.

Principles and Biomedical Application of Graphene Family Nanomaterials.

Two-dimensional graphene family nanomaterials (GFNs) are extensively studied by the researchers for their quantum size effect, large surface area, numerous reactive functional sites, and biocompatibility. The hybrid materials of GFNs exhibit an increased level of mechanical strength, optical, electronic, and catalytic activity due to their incorporation. The application of GFNs in the energy, environment, electric and electronic, personal care, and health sectors is abundant, which is not only by their unique physicochemical properties but also by their ease and large production by various synthetic approaches and economically inexpensiveness. Their general biomedical applications include bioimaging, biosensing, drug delivery, tissue engineering, killing the microbes, and demolishing the cancer tumor. The first chapter of this book describes definitions, synthetic methods, unique properties, and biomedical applications of GFNs, including graphene, graphene oxide, and reduced graphene oxide.

Role of Graphene Family Nanomaterials in Skin Wound Healing and Regeneration.

Owing to astonishing properties such as the large surface area to volume ratio, mechanical stability, antimicrobial property, and collagen crosslinking, graphene family nanomaterials (GFNs) have been widely used in various biomedical applications including tissue regeneration. Many review literatures are available to compile the role of GFNs in cardiac, bone, and neuronal tissue regeneration. However, the contribution of GFNs in skin wound healing and tissue regeneration was not yet discussed. In the present review, we have highlighted the properties of GFNs and their application in skin wound healing. In addition, we have included challenges and future directions of GFNs in skin tissue regeneration in the portion of conclusion and perspectives.

Graphene for Nanobiosensors and Nanobiochips.

Graphene family nanomaterials have interesting electronic structures which determine their electrical, optical, and mechanical properties. Especially, their unique chemical properties enable interactions with biological substances and chemical reagents, and the interactions have further an influence on the observable properties of the graphene family nanomaterials. Such aspects render graphene family nanomaterials versatile for various types of biosensing as a target recognition unit and a recognition-to-signal transduction unit. In this chapter, we look over the recent progress on the graphene-based biosensors, which is categorized in terms of (1) the role of graphene family nanomaterials (target recognition, signal transduction), (2) the sensing mechanisms and modes (electrochemical, electrical, fluorescent, Raman scattering), and (3) the formats of sensing devices (paper, lab-on-a-chip, wearable devices).

Reflections and Outlook on Multifaceted Biomedical Applications of Graphene.

Two-dimensional nanomaterials have been widely explored by researchers due to their nanosized thickness and quantum size effect. They were layered double hydroxides, transition metal dichalcogenides, transition metal oxides, and synthetic silicate clays. Among the 2D nanomaterials, graphene and their derivatives were investigated extensively at first as they exhibited exceptional conductivity and a zero-band gap semimetal nature. Though graphene family nanomaterials (GFNs) were utilized for several physicochemical applications, including electronic, electric, mechanic, photonic, magnetic, and catalytic devices, their biomedical applications are still meritorious. Biosensor, bioimaging, drug delivery, tumor ablation, and tissue regeneration are some of them. The outlook of the present book chapters encompasses the preparation of GFNs, physicochemical properties, biomedical applications, biosafety, and their future directions.

Graphene Family Nanomaterials (GFN)-TiO2 for the Photocatalytic Removal of Water and Air Pollutants: Synthesis, Characterization, and Applications.

Given the industrial revolutions and resource scarcity, the development of green technologies which aims to conserve resources and reduce the negative impacts of technology on the environment has become a critical issue of concern. One example is heterogeneous photocatalytic degradation. Titanium dioxide (TiO2) has been intensively researched given its low toxicity and photocatalytic effects under ultraviolet (UV) light irradiation. The advantages conferred by the physical and electrochemical properties of graphene family nanomaterials (GFN) have contributed to the combination of GFN and TiO2 as well as the current variety of GFN-TiO2 catalysts that have exhibited improved characteristics such as greater electron transfer and narrower bandgaps for more potential applications, including those under visible light irradiation. In this review, points of view on the intrinsic properties of TiO2, GFNs (pristine graphene, graphene oxide (GO), reduced GO, and graphene quantum dots (GQDs)), and GFN-TiO2 are presented. This review also explains practical synthesis techniques along with perspective characteristics of these TiO2- and/or graphene-based materials. The enhancement of the photocatalytic activity by using GFN-TiO2 and its improved photocatalytic reactions for the treatment of organic, inorganic, and biological pollutants in water and air phases are reported. It is expected that this review can provide insights into the key to optimizing the photocatalytic activity of GFN-TiO2 and possible directions for future development in these fields.

Direct and Indirect Genotoxicity of Graphene Family Nanomaterials on DNA-A Review.

Graphene family nanomaterials (GFNs), including graphene, graphene oxide (GO), reduced graphene oxide (rGO), and graphene quantum dots (GQDs), have manifold potential applications, leading to the possibility of their release into environments and the exposure to humans and other organisms. However, the genotoxicity of GFNs on DNA remains largely unknown. In this review, we highlight the interactions between DNA and GFNs and summarize the mechanisms of genotoxicity induced by GFNs. Generally, the genotoxicity can be sub-classified into direct genotoxicity and indirect genotoxicity. The direct genotoxicity (e.g., direct physical nucleus and DNA damage) and indirect genotoxicity mechanisms (e.g., physical destruction, oxidative stress, epigenetic toxicity, and DNA replication) of GFNs were summarized in the manuscript, respectively. Moreover, the influences factors, such as physicochemical properties, exposure dose, and time, on the genotoxicity of GFNs are also briefly discussed. Given the important role of genotoxicity in GFNs exposure risk assessment, future research should be conducted on the following: (1) developing reliable testing methods; (2) elucidating the response mechanisms associated with genotoxicity in depth; and (3) enriching the evaluation database regarding the type of GFNs, applied dosages, and exposure times.

Graphene Oxide and Reduced Graphene Oxide Exhibit Cardiotoxicity Through the Regulation of Lipid Peroxidation, Oxidative Stress, and Mitochondrial Dysfunction.

OBJECTIVE: Graphene has been widely used for various biological and biomedical applications due to its unique physiochemical properties. This study aimed to evaluate the cardiotoxicity of graphene oxide (GO) and reduced GO (rGO) in vitro and in vivo, as well as to investigate the underlying toxicity mechanisms. METHODS: GO was reduced by gamma irradiation to prepare rGO and then characterized by UV/visible light absorption spectroscopy. Rat myocardial cells (H9C2) were exposed to GO or rGO with different absorbed radiation doses. The in vitro cytotoxicity was evaluated by MTT assay, cell apoptosis assay, and lactate dehydrogenase (LDH) activity assay. The effects of GO and rGO on oxidative damage and mitochondrial membrane potential were also explored in H9C2 cells. For in vivo experiments, mice were injected with GO or rGO. The histopathological changes of heart tissues, as well as myocardial enzyme activity and lipid peroxidation indicators in heart tissues were further investigated. RESULTS: rGO was developed from GO following different doses of gamma irradiation. In vitro experiments in H9C2 cells showed that compared with control cells, both GO and rGO treatment inhibited cell viability, promoted cell apoptosis, and elevated the LDH release. With the increasing radiation absorbed dose, the cytotoxicity of rGO gradually increased. Notably, GO or rGO treatment increased the content of ROS and reduced the mitochondrial membrane potential in H9C2 cells. In vivo experiments also revealed that GO or rGO treatment damaged the myocardial tissues and changed the activities of several myocardial enzymes and the lipid peroxidation indicators in the myocardial tissues. CONCLUSION: GO exhibited a lower cardiotoxicity than rGO due to the structure difference, and the cardiotoxicity of GO and rGO might be mediated by lipid peroxidation, oxidative stress, and mitochondrial dysfunction.

Injectable in situ forming thermo-responsive graphene based hydrogels for cancer chemo-photothermal therapy and NIR light-enhanced antibacterial applications.

Functionalized graphene oxide (GO) and reduced GO (rGO) based nanomaterials hold a great potential for cancer photothermal therapy. However, their systemic administration has been associated with an accelerated blood clearance and/or with suboptimal tumor uptake. To address these limitations, the local delivery of GO/rGO to the tumor site by 3D matrices arises as a promising strategy. In this work, injectable chitosan-agarose in situ forming thermo-responsive hydrogels incorporating GO (thermogel-GO) or rGO (thermogel-rGO) were prepared for the first time. The hydrogels displayed suitable injectability and gelation time, as well as good physicochemical properties and cytocompatibility. When irradiated with near infrared (NIR) light, the thermogel-rGO produced a 3.8-times higher temperature increase than thermogel-GO, thus decreasing breast cancer cells' viability to 60%. By incorporating an optimized molar ratio of the Doxorubicin:Ibuprofen combination on thermogel-rGO, this formulation mediated a chemo-photothermal effect that further diminished cancer cells' viability to 34%. In addition, the hydrogels' antibacterial activity was further enhanced upon NIR laser irradiation, which is an important feature considering the possible risk of infection at the site of administration. Overall, thermogel-rGO is a promising injectable in situ forming hydrogel for combinatorial chemo-photothermal therapy of breast cancer cells and NIR light enhanced antibacterial applications.

Toxicology data of graphene-family nanomaterials: an update.

Due to its unique physical structure and chemical properties, graphene family nanomaterials (GFNs) and derived commodities have been widely used in commercial products, particularly biomedical applications, which has significantly increased the risk of human exposure. There exists significant evidence that GFNs are accumulated in a number of tissues and organs through different exposure pathways, and further cause toxicity manifested as lesions or functional impairment. Moreover, GFNs can be internalized by varing cell types and induce cytoskeletal disorders, organelle dysfunction, and interact directly with biological macromolecules such as DNA, mRNA and proteins, ultimately resulting in greater rates of cell apoptosis, necrosis and autophagic cell death. The toxicological effect of GFN is closely related to its lateral size, surface structure, functionalization, and propensity to adsorb proteins. Using major data published over the past four years, this review presents and summarizes state of current understanding of GFN toxicology and identifies current deficiencies and challenges. This review aims to help improve evaluation of the biocompatibility of GFNs and provides theoretical guidance for their safe application.

Interaction of graphene-family nanomaterials with microbial communities in sequential batch reactors revealed by high-throughput sequencing.

The accelerated development and application of graphene-family nanomaterials (GFNs) have increased their release to various environments and converged in wastewater treatment plants (WWTPs). However, little is known about the interactions between GFNs and microbes in WWTPs. In this study, the interaction of graphene oxide (GO) or graphene (G) at different concentrations with microbial communities in sequential batch reactors was investigated. Transmission electron microscopy and Raman spectroscopy analyses showed that the structures of GFNs were obviously changed, which suggested GFNs could be degraded by some microbes. Significantly higher DNA concentration and lower cell number in high-concentration GO group were detected by DNA leakage test and qPCR analysis, which confirmed the microbial toxicity of GO. The chemical oxygen demand and ammonia nitrogen removals were significantly affected by G and GO with high concentrations. Further, high-throughput sequencing confirmed the composition and dynamic changes of microbial communities under GFNs exposure. Saccharibacteria genera incertae sedis (12.55-28.05%) and Nakamurella (20.45-29.30%) were the predominant genera at two stages, respectively. FAPROTAX suggested 12 functional groups with obvious changes related to the biogeochemical cycle of C, N and S. Molecular ecological network analysis showed that the networks were more complex in the presence of GFNs, and the increased negative interactions reflected more competition relationships in microbial communities. This study is the first to report the effect of GFNs on network of microbial communities, which provides in-depth insights into the complex and highlights concerns regarding the risk of GFNs to WWTPs.

Aqueous aggregation and stability of graphene nanoplatelets, graphene oxide, and reduced graphene oxide in simulated natural environmental conditions: complex roles of surface and solution chemistry.

Graphene-family nanomaterials (GFNs) exhibit universal applications and consequently will inevitably enter aquatic systems. However, both the fate and behavior of GFNs in aquatic environments have not been completely explored at field relevant conditions. Herein, we have systematically investigated the aqueous aggregation and stability of graphene nanoplatelets (GNPs), graphene oxide (GO), and reduced graphene oxide (RGO) under varied solution chemistry parameters (pH, divalent cations, and dissolved organic carbon (DOC)) during 21 days of incubation in simulated natural environmental conditions. Results indicate that pH values from 6 to 9 had a notable impact on the aqueous behaviors of the three GFNs. Divalent cations (Ca2+ and Mg2+) at the concentrations of 2.5 and 10 mM remarkably increased the extent of aggregation of the three GFNs and resulted in severe sedimentation, independently of surface chemical functionalization. The presence of only DOC ranging from 0.5 to 2 mg C/L significantly elevated the dispersion stability of GNPs and RGO in a dose-dependent manner, whereas no effects were observed on GO. Furthermore, DOC at the studied concentrations and surface functionality were insufficient to counterbalance the impact of the divalent cations. Direct visual and in situ observations further supported the conclusions on the effects of divalent cations or/and DOC. These findings further underline that the environmental behaviors of GFNs are controlled by the complex interplay between water chemistry parameters and GFN surface properties.