Assessment of acute neurotoxicity of nitrogen-doped multilayer graphene nanoparticles and their capability to change Cd2+/Pb2+/Hg2+-induced injury in brain cortex nerve terminals

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Palladin Institute of Biochemistry of the National Academy of Sciences of Ukraine, Kyiv

Pisarzhevskii Institute of Physical Chemistry of the National Academy of Sciences of Ukraine, Kyiv

Assessment of acute neurotoxicity of nitrogen-doped multilayer graphene nanoparticles and their capability to change Cd2+/Pb2+/Hg2+-induced injury in brain cortex nerve terminals

Marina Dudarenko

Nataliya Krisanova

Natalia Pozdnyakova

Artem Pastukhov

Olena Pariiska

Yaroslav Kurys

Svitlana Sotnik

Sergey V. Kolotilov

Vyacheslav Koshechko

Tatiana Borisova

Abstract

neurotoxic risk nerve terminal

Graphene materials are widely used in different technologies and certainly released into aquatic and air surroundings being environmental pollution components. Nitrogen-doped graphene nanomaterials have great potential for application, in particular, in energy storage, as electrochemical sensors and waste water treatment. The neurotoxic risk of nitrogen-doped multilayer graphene is unknown.

Aim. To evaluate neurotoxic risk of nitrogen-doped multilayer graphene.

Methods. Here, nitrogen-doped multilayer graphene nanoparticles (N-MLG) were synthesized by means of electrochemical exfoliation of high-purity graphite rods in NaN3-based electrolyte and characterised using TEM, AFM and UV-vis spectroscopy. Neuroactive features of N-MLG were assessed in isolated cortex nerve terminals (synaptosomes) analysing the extracellular level of excitatory neurotransmitter L-[14C] glutamate and inhibitory one [3H]GABA.

Results. It was revealed that N-MLG did not affect the extracellular synaptosomal levels of L-[14C]glutamate and [3H]GABA within the concentration range 0.01-0.5 mg/ml, and an increase in a concentration up to 1 mg/ml caused an insignificant increase (tendency to increase) in these levels for both neurotransmitters. To analyse a capability of interaction with heavy metals in biological system, N-MLG was investigated using model of acute Cd2+/Pb2+/Hg2+-induced neurotoxicity in nerve terminals. It was revealed that Cd2+/Pb2+/Hg2+-induced increase in the extracellular level of L-[14C] glutamate and [3H]GABA was not changed by N-MLG.

Conclusions. N-MLG does not possess neurotoxic signs and is biocompatible within the concentration range 0.01-1 mg/ml. In biological system, N-MLG did not mitigate/aggravate Cd2+/Pb2+/Hg2+- induced neurotoxicity in nerve terminals.

Key words: nitrogen-doped multilayer graphene; nanoparticles; heavy metals; neurotoxicity; glutamate; GABA; brain nerve terminals.

Graphene is a carbon 2D crystal consisting of a two dimensional honeycomb-lattice structure made of sp2-hybridized carbon atoms. Graphene and its derivatives due to their two-dimensional shape and special mechanical, electronic, optical and catalytic properties are widely used in diverse fields, e.g. nanoelectronics, catalysis, and water technologies. Graphene is considered one of the most promising nanomaterials for biomedical applications and nanomedicine [1, 2]. Nitrogen-doped graphene exhibits excellent electrochemical parameters and has great potential for applications in energy storage, as a high-performance catalyst support for fuel cell electrocatalysis and sensor electrochemical applications [2, 3]. Nitrogen-doped graphene nanomaterials are effective for removing toxic species, e.g. organic pollutants, dyes, heavy metal ions from wastewater [4, 5]. In particular, nitrogen-doping enhances the interaction between the active sites and organic or ionic species, thereby improving removal efficiency of these pollutants [2].

Graphene materials during production and application can find ways into the environment in the form of nanoparticles because of its high dispersity in many solvents [6, 7]. In this context, graphene materials can widely pollute water resources and air around the world.

Nowadays, it is clear that carbonaceous airborne particulate matter is the most abandon environmental pollutant that can reach the nervous system of humans and trigger development of neurological and neurodegenerative disorders/diseases [810]. Expansion of these disorders/diseases is one of the main reasons of disability and premature death in Europe and linked to air pollution with particulate matter [8, 11-13]. From one hand, particulate matter can reach human organism through lungs and gut to the circulatory system and possibly cross the blood-brain barrier [14-18]. From the other hand, particulate matter can get to the human nervous system along the olfactory axon and overcome the blood-brain barrier [19, 20]. Recently, we have characterised neuroactivity of carbon-based nanoparticles, such as carbon nanodots, nanodiamonds, fullerene C60, carbonaceous smoke particulate matter from combustion of plastics and wood samples. Transport kinetics of key excitatory neurotransmitters, L-[14C] glutamate and inhibitory one [3H] GABA, were changed by above nanoparticles that in turn can provoke presynaptic malfunction and development of neuropathology [21, 22]. Carbonaceous airborne particulate matter possesses adhesive surface and can bind potentially toxic molecules, including heavy metals [23, 24].

The neurotoxic potential of nitrogen-doped multilayer graphene is unknown. In neurobiology study of other graphene derivatives, it was demonstrated that few-layer pristine graphene and monolayer graphene oxide flakes were in contact with the neuronal membrane and free in the cytoplasm, but did not have significant impact on neuron viability. Graphene oxide exposure inhibited excitatory transmission, reduced the number of excitatory synaptic contacts, and concomitantly enhanced the inhibitory activity. This was accompanied by altered Ca2+ dynamics in excitatory and inhibitory neurons. So, it was suggested that both graphene preparations affected neuronal transmission [1]. Taking into account abovementioned facts, the aims of the present study were (*) to synthesize nitrogen-doped multilayer graphene nanoparticles (N-MLG) by means of electrochemical exfoliation of high-purity graphite rods; (**) to assess neuromodulatory features of N-MLG in isolated presynaptic rat cortex nerve terminals (synaptosomes) analysing the extracellular level of neurotransmitters L-[14C] glutamate and [3H]GABA; and (***) to examine the effects of N-MLG on the neurotoxic injury caused by heavy metal ions Cd2+, Pb2+, and Hg2+ in the nerve terminals.

Materials and methods

HEPES, EGTA, EDTA, NaN3, Ficoll 400, High Performance LSC Cocktail salts of the analytical grade were obtained from Sigma, USA; L-[14C] glutamate; [3H] GABA-Perkin Elmer, Waltham, MA, uSa. The high-purity graphite rods (99.9995%) were obtained from Alfa Aesar.

Transmission electron microscopy (TEM) images of the synthesized graphene material were recorded using a microscope TEM125K (Selmi) with an accelerating voltage 100 kV. Atomic force microscopy (AFM) of thin film N-MLG samples on the surface of silicon wafers coated with silicon nitride (Agar Scientific) was performed on a Nanoscope IIIa Dimension 3000TM (Digital) instrument. UV-vis-spectra of N-MLG dispersions were registered via UV- visible spectrometer 4802 (Unico).

Synthesis of multilayer graphene doped with nitrogen

Multilayer graphene doped with nitrogen (N-MLG) was synthesized according to previous published procedure by means of electrochemical exfoliation of high-purity graphite rods (Alfa Aesar, 99.9995%) in 1.0 M aqueous solution of NaN3 using three-electrode undivided cell and potentiostat PI-50-1.1 [25]. Graphite rods were used as working and auxiliary electrodes and Ag/AgCl as reference electrode. Exfoliation procedure was carried out during 20 h via polarization of graphite electrode by +4.0 V (50 s) and 0.0 V (50 s) with multiple changing of polarization potential (pulse mode of electrolysis).

N-MLG particles from the obtained aqueous dispersion were separated on a nylon membrane filter with a pore diameter 0.2 pm (SUPELCO°), rinsed with water, and dried in oven at 60 °C. If necessary, the regeneration of N-MLG dispersion was performed by shortterm ultrasound treatment (2 min) of the dried graphene material in appropriate solvent (for example, water) using the ultrasonic washing bath.

Animals and Ethics

Wistar rats (males), 3 months' age, were kept in the vivarium of Palladin Institute of Biochemistry, NAS of Ukraine in a quiet and temperature-controlled room at 22-23 °C. Animals were provided with dry food pellets and water ad libitum. All animal-involving procedures were performed in accordance with the guidelines of the European Community (2010/63/EU); “Scientific Requirements and Research Protocols”; “Research Ethics Committees” of Declaration of Helsinki; and “ARRIVE guidelines for reporting experiments involving animal” [26, 27]; and also local Ukrainian laws and policies. The experimental protocols were approved by the Animal Care and Use Committee of Palladin Institute of Biochemistry (Protocol # 1 from 10/01/2023). The total number of animals was 9.

Isolation of the synaptosomes from the rat cortex

The synaptosomes represented ~87% of the particles in electron micrographs of preparations, and did not contain nerve cell bodies and functional glial fragments [28-30]. Nerve terminals were isolated from the cortex regions of rat brain. Isolation procedures were conducted at + 4 °C. The cortex regions were removed and homogenized in the following ice-cold solution: sucrose 0.32 M; HEPES-NaOH 5 mM, pH 7.4; EDTA 0.2 mM. One synaptosome preparation was obtained from one rat. The synaptosomes were isolated according to Cotman method with minor modifications [9, 31-33] by differential/ Ficoll-400 density gradient centrifugation. The synaptosomes were fitting for experiments for 2-4 hours after isolation. The standard saline solution contained: NaCl 126 mM; KCl 5 mM; MgCl2 2.0 mM; NaH2PO4 1.0 mM; HEPES 20 mM, pH 7.4; and D-glucose 10 mM. Protein concentrations were monitored according to Larson [34].

The extracellular synaptosomal level of L-[14C] glutamate

The synaptosomes were diluted up to a concentration of 2 mg of protein/ml, and then the synaptosomes were pre-incubated at 37 °C for 10 min, and loaded with L-[14C] glutamate, 1 nmol per mg of protein, 238 mCi/mmol, at 37 °C for 10 min. The synaptosomes after loading were washed with 10 volumes of the ice-cold standard saline solution and centrifuged (10,000xg, 20 s) at +4 °C; the pellets were re-suspended in the standard saline solution up to 1 mg protein/ml. The extracellular L-[14C] glutamate level was assessed in the synaptosome suspensions (125 pl, 0.5 mg of protein/ml). The synaptosome aliquots were preincubated for 8 min to restore the ion gradients, and then Cd2+ (1 mM CdCL), Pb2+ (2.5 mM Pb acetatate (PbAc)), and Hg2+ (2.5 pM HgCl2) were added to the synaptosomes and they were further incubated at 37 °C during 0 and 6 min; then centrifuged at 10,000 x g for 20 s at room temperature. The values of L-[14C] glutamate release were monitored in the supernatant aliquots (100 pl), and the pellets preliminary treated with SdS (100 pl of 10% SDS stock solution) by liquid scintillation counting using the scintillation cocktail ACS (1.5 ml) and liquid scintillation counter Hidex 600SL (Finland) [35]. The experimental data were collected from “n” independent experiments carried out in triplicate using different synaptosome preparations.

The extracellular synaptosomal level of [3H] GABA

The synaptosomes were diluted up to 2 mg of protein/ml; after pre-incubation at 37 °C for 10 min, the synaptosomes were preloaded with [3H]GABA (50 nM, 4.7 pCi/ml) in the standard saline solution at 37 °C for 10 min. Aminooxyacetic acid (100 pM) was added to the incubation medium throughout [3H] GABA experiments. After pre-loading, the suspension was washed with 10 volumes of the ice-cold standard saline solution. The pellets were re-suspended in the standard saline solution up to 1 mg of protein/ml. The synaptosome aliquots were pre-incubated for 8 min to restore the ion gradients, and then Cd2+ (1 mM CdCl2), Pb2+ (1 mM PbAc) and Hg2+ (2.5 pM HgCl2) were added and further incubated at 37 C during 0 and 5 min; then centrifuged at 10,000 x g for 20 s at room temperature [36]. [3H] GABA was measured in the supernatant aliquots (90 ці) by liquid scintillation counting with Sigma-Fluor® High Performance LSC Cocktail (1.5 ml) using liquid scintillation counter Hidex 600SL (Finland), and the extracellular level was expressed as the percentage of total accumulated [3H] GABA [37]. The data were collected from “n” independent experiments performed in triplicate with different synaptosome preparations.

Statistical analysis

The experimental data were expressed as the mean ± S.E.M. of n independent experiments. One-way and two-way ANOVA were applied; the accepted significance level was P < 0.05. Two-way ANOVA followed by Tukey's post hoc test was applied to assess the interactions between N-MLG and Cd2+, Pb2+, and Hg2+ (N-MLG treatment and Cd2+/Pb2+/ Hg2+ treatment were the independent factors).

Results and discussion

Characterization of multilayer graphene doped with nitrogen

During pulse mode of electrolysis, multiple (cyclic) azide anions (N3-- intercalation/ deintercalation into graphite interlayer space occurs, also associated with N2, NH3 and O2 evolution due to partial anodic/cathodic decomposition of N3-. This provides exfoliation of the graphite electrode, forming multilayer packages of graphene, doped with nitrogen, and its transition in the electrolyte volume [25].

The obtained graphene material was characterized by several instrumental analysis methods. On the TEM images of N-MLG (Fig. 1, A) graphene sheets with lateral size from several hundred nanometers to several microns, forming multiple-layered lamellar structures, can be observed.

According to the data of AFM experiment, shown on the Fig. 1b, the lateral size of N-MLG particles, taken from a dilute dispersion in ethanol, is consistent with the data obtained by TEM. From the crosssectional profile analysis (Fig. 1, B) it can be seen that the thickness of the N-MLG particles present in the AFM image is 4.2 nm. Earlier, on the basis of AFM data, we showed that the thickness of the monolayer in the N-MLG was 0.6 nm [25]. Taking this into account, it can be assumed that the obtained N-MLG particles are multilayered, consisting of the packages of ~7 graphene layers. At the same time, we cannot rule out the presence of a small number of N-MLG particles of both smaller (< 7 graphene layers) and larger (up to 9 graphene layers) thicknesses obtained by the electrochemical approach used herein [25]. Thus, AFM data also confirm the multilayered nature of the obtained graphene material.

UV-vis spectrum of N-MLG dispersions in ethanol is provided on Fig. 2. The presence of a band with an absorption maximum at 267 nm, corresponding to the %%* transition in the C=C bonds of graphene, indicates in favor of a slight oxidation of the resulting N-MLG.

Neurotoxicity studies using rat brain nerve terminals

The extracellular levels of neurotransmitters glutamate and GABA are essential synaptic characteristics which show a balance of transporter-mediated uptake and tonic release of neurotransmitters [38, 39].

In the first series of the experiments, the effects of N-MLG on the extracellular levels of excitatory neurotransmitter L-[14C] glutamate and inhibitory one [3H]GABA were assessed in the nerve terminals. It was revealed that N-MLG did not affect the extracellular levels of L-[14C] glutamate and [3H]GABA within the concentration range 0.01-0.5 mg/ml, while an increase in a concentration up to 1 mg/ml caused an insignificant elevation (tendency to increase) of these levels for both neurotransmitters (Fig. 3). Therefore, N-MLG did not possess neurotoxic signs and was biocompatible within the concentration range 0.01-1 mg/ml.

In the next series of the experiments, it was investigated whether N-MLG could modulate a Cd2+/Pb2+/Hg2+-induced increase in the extracellular levels of L-[14C] glutamate and [3H]GABA in nerve terminals. Based on above data, a N-MLG concentration of 0.5 mg/ml was chosen for further experiments with heavy metals. It was found that N-MLG (0.5 mg/ ml) did not change Cd2+/Pb2+/Hg2+-induced increase in the extracellular levels of L-[14C] glutamate and [3H]GABA in nerve terminals (Fig. 4), and so N-MLG was inert regarding modulation/aggravation of Cd2+/Pb2+/Hg2+- induced neurotoxic effects in nerve terminals.

Two-way ANOVA revealed no interaction between Cd2+ and N-MLG [F(1,32) = 0.009; p = 0.92; n = 9], between Pb2+ and N-MLG [F(1 32) = 1.55; p = 0.22; n = 9] and between Hg2+ and N-MLG [F(1,32) = 1.22; p = 0.27; n = 9] in L-[14C] glutamate experiments.

Two-way ANOVA revealed no interaction between Cd2+ and N-MLG [F(1 32) = 0.69; p = 0.41; n = 9], between Pb2+ and N-MLG [F(1,32) = 0.22; p = 0.63; n = 9] and between Hg2+ and N-MLG [F(1,32) = 0.13; p = 0.72; n = 9] in [3H] GABA experiments.

Fig. 1. TEM image (A) and AFM image with corresponding cross-sectional profile (B) of N-MLG particles

Fig. 2. UV-vis spectra of N-MLG dispersion in ethanol

Fig. 3. The extracellular levels of neurotransmitters L-[14C] glutamate (A) and [3H]GABA (B) in the nerve terminals in the presence of N-MLG (0.01-1.0 mg/ml)

Data are the mean ± SEM. n.s., no significant differences as compared to the appropriate control, n = 9

Graphene and its derivatives have a potential to make a very significant impact on society with applications in the biomedical field. A possibility to engineer graphene-based medical devices at the neuronal interface is of particular interest, making it imperative to determine the biocompatibility of graphene materials with neuronal cells [1]. However, wide production and application of neuroactive graphene and its derivatives can increase uncontrolled environment pollution with these nanoparticles increasing risks to nervous system. To avoid the problem, technological- oriented graphene and its derivatives should be analysed regarding their neurotoxicity as single pollutants and also in combination with other pollutants, e.g. heavy metals, using multipollutant approach.

Here, we showed that N-MLG did not affect the extracellular synaptosomal levels of L-[14C] glutamate and [3H]GABA within the concentration range 0.01-1 mg/ml. Therefore, N-MLG can be applied in different technologies, including neurotechnologies and waste water treatment, because N-MLG did not possess neurotoxic signs and is biocompatible within this concentration range and it is safe when become environmental pollution component. Among other carbon- based nanoparticles that we have investigated regarding changes in the extracellular levels of L-[14C] glutamate and [3H]GABA in nerve terminals, N-MLG was less neurotoxic as compared to carbon dots [40], nanodiamonds [35], fullerene C60 [41], plastic and wood smoke particulate matter [21, 22, 42]. In this contex, N-MLG is more promising for neurotechnologies then other carbon nanoparticles because of absence of neurotoxic signs and its neurocompatibility.

Literature data have demonstrated that due to weak hydrophobic interactions, the negatively charged surfaces of graphene oxide were favorable to interact through electrostatic attractions with organic and inorganic cations [43, 44]. Graphene oxide could remove heavy metal ions as carriers due to its large surface area, pore size and abundant oxygen-containing functional groups [45-47]. In the present study, we revealed that N-MLG did not change Cd2+/Pb2+/Hg2+-induced increase in the extracellular level of L-[14C] glutamate and [3H]GABA and so the nanoparticles did not mitigate/aggravate Cd2+/Pb2+/Hg2+- induced neurotoxicity in nerve terminals. It also means that despite potential application in technologies, in particulate in waste water treatment against heavy metals, N-MLG was not effective against prevention/mitigation of Cd2+/Pb2+/Hg2+-induced neurotoxicity in biological system such as nerve terminals.

Conclusions

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Fig. 4. Effects of N-MLG (0.5 mg/ml) on Cd2+/Pb2+/Hg2+-induced increase in the extracellular levels of L-[14C] glutamate (a) and [3H]GABA (b)

Data are the mean ± SEM. ***, P < 0.001; as compared to the control; n.s., no significant differences as compared to Cd2+/Pb2+/Hg2+ effects, n = 9

N-MLG, prepared by exfoliation of the graphite electrode in azide-containing electrolyte, did not influence the extracellular levels of L-[14C] glutamate and [3H] GABA in the nerve terminals. Cd2+/Pb2+/Hg2+-induced increase in the extracellular levels of L-[14C] glutamate and [3H]GABA was not changed by N-MLG that was shown using model of acute neurotoxicity in nerve terminals. Therefore, N-MLG did not possess neurotoxic signs and is neurocompatible. In biological system, N-MLG did not mitigate or aggravate Cd2+/Pb2+/Hg2+-induced neurotoxicity in nerve terminals.

Competing interests

The authors declare no financial and non- financial competing interests exist.

Authors contributions

N-MLG synthesis and characterisation -- OP, YK; synaptosome preparations were obtained by aP & MD, L-[14C] glutamate experiments -- AP & KN, [3H] GABA experiments -- NP & MD; data analysis and figure preparation -- OP, YK, SS, NP, KN, TB, SK; funding acquisitions, project leading, data analysis and paper writing -- TB, VK.

Ethical Approval

Animal-involved experiments were performed according to the “Scientific Requirements and Research Protocols” & “Research Ethics Committees” of Declaration of Helsinki; the “ARRIVE guidelines for reporting experiments involving animal” [26, 27], and were preliminary approved by Institutional Animal Care and Use Committee (Protocol # 1 from 10/01/2023).

Consent to Participate

Not applicable.

Consent to Publish

Not applicable.

Availability of data and materials

All data generated or analyzed during this study are included in this published article.

Acknowledgements and Funding

This work was supported by the National Research Foundation of Ukraine: NRFU grant #2021.01/0061 «Cumulative neurotoxic effect of multicomponent pollution by airborne particles and neuroactive pharmaceuticals, biomaterials (including SARS-CoV-2), toxic metals and its prevention», PI of the Project: Prof. T. Borisova.

References

1. Bramini M., Sacchetti S,ArmirottiA., Rocchi A., Vazquez E., Leon Castellanos V., Bandiera T., Cesca F., Benfenati F. Graphene Oxide Nanosheets Disrupt Lipid Composition, Ca 2+ Homeostasis, and Synaptic Transmission in Primary Cortical Neurons. ACS Nano. 2016, 10 (7), 7154-7171. https://doi.org/10.1021/acsnano.6b03438.

2. Yokwana K., Ntsendwana B., Nxumalo E.N., Mhlanga S.D. Recent advances in nitrogen-doped graphene oxide nanomaterials: Synthesis and applications in energy storage, sensor electrochemical applications and water treatment. 2023, 38 (13), 3239-3263. https://doi.org/10.1557/S43578-023-01070-1.

3. Xiong B., Zhou Y., Zhao Y., Wang J., Chen X., O'Hayre R., Shao Z. The use of nitrogen-doped graphene supporting Pt nanoparticles as a catalyst for methanol electrocatalytic oxidation. Carbon N Y. 2013, 52 (52), 181-192. https://doi.org/10.1016/j.carbon.2012.09.019.

4. Chen F., Guo L., Zhang X., Leong Z.Y., Yang S., Yang H.Y. Nitrogen-doped graphene oxide for effectively removing boron ions from seawater. Nanoscale. 2016, 9 (1), 326-333. https://doi.org/10.1039/C6NR07448K.

5. Liu Y., Liu H., Zhou Z., Wang T., Ong C.N., Ve- citis C.D. Degradation of the Common Aqueous Antibiotic Tetracycline using a Carbon Nanotube Electrochemical Filter. Environ Sci Technol. 2015, 49 (13), 7974-7980. https://doi.org/10.1021/ACS.EST.5B00870.

6. Zhao J., Wang Z., White J.C., Xing B. Graphene in the aquatic environment: Adsorption, dispersion, toxicity and transformation. Environ Sci Technol. 2014, 48 (17), 9995-10009. https://doi.org/10.1021/ES5022679.

7. Bussy C., Ali-Boucetta H., Kostarelos K. Safety Considerations for Graphene: Lessons Learnt from Carbon Nanotubes. Acc Chem Res. 2013, 46 (3), 692-701. https://doi.org/10.1021/ar300199e.

8. Landrigan P.J., Fuller R., Acosta N.J.R.R., Adeyi O., Arnold R., Basu N. (Nil)., Balde A.B., Bertollini R., Bose-O'Reilly S., Boufford J.I., Breysse P.N., Chiles T., Mahidol C., Coll-Seck A.M., Cropper M.L., Fobil J., Fuster V., Greenstone M., Haines A., Hanrahan D., Hunter D., Khare M., Krupnick A., Lanphear B., Lohani B., Martin K., Mathiasen K.V., McTeer M.A., Murray C.J.L.L., Ndahimananjara J.D., Perera F., Potocnik J., Preker A.S., Ramesh J., Rockstrom J., Salinas C., Samson L.D., Sandilya K., Sly P.D., Smith K.R., Steiner A., Stewart R.B., Suk W.A., van Schayck O.C.P.P., Yadama G.N., Yumkella K., Zhong M. The Lancet Commission on pollution and health. Lancet. 2018, 391 (10119), 462-512. https://doi.org/10.1016/S0140-6736(17)32345-0.

9. Borisova T. Nervous System Injury in Response to Contact With Environmental, Engineered and Planetary Micro- and NanoSized Particles. Front Physiol. 2018, 9, 728. https://doi.org/10.3389/fphys.2018.00728.

10. Tranvik L.J. New light on black carbon. Nat Geosci. 2018, 11 (8), 547-548. https://doi. org/10.1038/s41561-018-0181-x.

11. Deuschl G., Beghi E., Fazekas F., Varga T., Christoforidi K.A., Sipido E., Bassetti C.L., Vos T., Feigin V.L. The burden of neurological diseases in Europe: an analysis for the Global Burden of Disease Study 2017. Lancet Public Heal. 2020, 5 (10), e551-e567. https://doi.org/10.1016/S2468-2667(20)30190-0.

12. The Lancet Neurology. Long COVID: understanding the neurological effects. Lancet Neurol. 2021, 20 (4), 247. https://doi.org/10.1016/S1474-4422(21)00059-4.

13. Jarrahi A., Ahluwalia M., Khodadadi H., Da Silva Lopes Salles E., Kolhe R., Hess D.C., Vale F., Kumar M., Baban B., Vaibhav K., Dhandapani K.M. Neurological consequences of COVID-19: what have we learned and where do we go from here? J Neuroinflammation. 2020, 17 (1), 286. https://doi.org/10.1186/s12974-020-01957-4.

14. Jew K., Herr D., Wong C., Kennell A., Morris-Schaffer K., Oberdorster G, O'Banion MK., Cory-Slechta D.A., Elder A. Selective memory and behavioral alterations after ambient ultrafine particulate matter exposure in aged 3xTgAD Alzheimer's disease mice. Part Fibre Toxicol. 2019, 16 (1), 45. https://doi.org/10.1186/s12989-019-0323-3.

15. Morris-Schaffer K., Merrill A., Jew K., Wong C., Conrad K., Harvey K., Marvin E., Sobolewski M., Oberdorster G., Elder A., Cory-Slechta D. A. Effects of neonatal inhalation exposure to ultrafine carbon particles on pathology and behavioral outcomes in C57BL/6J mice. Part Fibre Toxicol. 2019, 16 (1), 10. https://doi.org/10.1186/s12989-019-0293-5.

16. Kim E.A. Particulate matter (fine particle) and urologic diseases. Int Neurourol J. 2017, 21 (3), 155-162. https://doi.org/10.5213/INJ.1734954.477.

17. Giadka A., Rymaszewska J., Zatonski T. Impact of air pollution on depression and suicide. Int J Occup Med Environ Health. 2018, 31 (6), 711-721. https://doi.org/10.13075/IJOMEH.1896.01277.

18. Klocke C., Allen J.L., Sobolewski M., Mayer-Proschel M., Blum J.L., Lauterstein D., Zelikoff J.T., Cory-Slechta D.A. Neuropatho-logical consequences of gestational exposure to concentrated ambient fine and ultrafine particles in the mouse. Toxicol Sci. 2017, 156 (2), 492-508. https://doi.org/10.1093/TOXSCI/KFX010.

19. Oberdorster G., Sharp Z., Atudorei V., Elder A., Gelein R., Kreyling W., Cox C. Translocation of Inhaled Ultrafine Particles to the Brain. Inhal Toxicol. 2004, 16 (6-7), 437-445. https:// doi.org/10.1080/08958370490439597.

20. Oberdorster G., Oberdorster E., Oberdorster J. Nanotoxicology: an emerging discipline evolving from studies of ultrafine particles. Environ Health Perspect. 2005, 113 (7), 823-39. https://doi.org/10.1289/ehp.7339.

21. Borysov A., Tarasenko A., Krisanova N., Pozdnyakova N., Pastukhov A., Dudarenko M., Paliienko K., Borisova T. Plastic smoke aerosol: Nano-sized particle distribution, absorption/fluorescent properties, dysregulation of oxidative processes and synaptic transmission in rat brain nerve terminals. Environ Pollut. 2020, 263, 114502. https://doi.org/10.1016/j.envpol.2020.114502.

22. Tarasenko A., Pozdnyakova N., Paliienko K., Borysov A., Krisanova N., Pastukhov A., Stanovyi O., Gnatyuk O., Dovbeshko G., Borisova T. A comparative study of wood sawdust and plastic smoke particulate matter with a focus on spectroscopic, fluorescent, oxidative, and neuroactive properties. Environ Sci Pollut Res Int. 2022, 29 (25), 38315-38330. https://doi.org/10.1007/s11356-022-18741-x.

23. Islam N., Dihingia A., Khare P., Saikia B.K. Atmospheric particulate matters in an Indian urban area: Health implications from potentially hazardous elements, cytotoxicity, and genotoxicity studies. J Hazard Mater. 2020, 384, 121472. https://doi.org/10.1016/j.jhazmat.2019.121472.

24. Islam N., Saikia B.K. An overview on atmospheric carbonaceous particulate matter into carbon nanomaterials: A new approach for air pollution mitigation. Chemosphere. 2022, 303, 135027. https://doi.org/10.1016/J.CHEMOSPHERE.2022.135027.

25. Ustavytska O., Kurys Y., Koshechko V., Pokhodenko V. One-Step Electrochemical Preparation of Multilayer Graphene Functionalized with Nitrogen. Nanoscale Res Lett. 2017, 12 (1), 175. https://doi.org/10.1186/ s11671-017-1957-4.

26. McGrath J., Drummond G., McLachlan E., Kilkenny C., Wainwright C. Guidelines for reporting experiments involving animals: the ARRIVE guidelines. Br J Pharmacol. 2010, 160 (7), 1573-1576. https://doi. org/10.1111/j.1476-5381.2010.00873.x.

27. Kilkenny C., Browne W., Cuthill I.C., Emerson M., Altman D.G. Animal research: Reporting in vivo experiments: The ARRIVE guidelines. Br J Pharmacol. 2010, 160 (7), 1577-1579. https://doi.org/10.1111/j.1476-5381.2010.00872.x.

28. Nicholls D.G. The glutamatergic nerve terminal. Eur J Biochem. 1993, 212 (3), 613-31. https://doi.org/10.1111/j.1432-1033.1993. tb17700.x.

29. Petr G.T., Sun Y., Frederick N.M., Zhou Y., Dhamne S.C., Hameed M.Q., Miranda C., Bedoya E.A., Fischer K.D., Armsen W., Wang J., Danbolt N.C., Rotenberg A., Aoki C.J., Rosenberg P.A. Conditional deletion of the glutamate transporter GLT-1 reveals that astrocytic GLT-1 protects against fatal epilepsy while neuronal GLT-1 contributes significantly to glutamate uptake into synaptosomes. J Neurosci. 2015, 35 (13), 51875201. https://doi.org/10.1523/JNEURO-SCI.4255-14.2015.

30. Gyorffy B.A., Kun J., Torok G., Bulyaki E., Borhegyi Z., Gulyassy P., Kis V., Szocsics P., Micsonai A., Matko J., Drahos L., Juhasz G., Kekesi K.A., Kardos J. Local apoptotic-like mechanisms underlie complementmediated synaptic pruning. Proc Natl Acad Sci USA. 2018, 115 (24), 6303-6308. https://doi.org/10.1073/pnas.1722613115.

31. Cotman C.W. Isolation of synaptosomal and synaptic plasma membrane fractions. Methods Enzymol. 1974, 31, 445-452.

32. Borisova T.A.,HimmelreichN.H. Centrifuge-induced hypergravity: [3H]GABA and l-[14C] glutamate uptake, exocytosis and efflux mediated by high-affinity, sodium-dependent transporters. Adv Sp Res. 2005, 36 (7), 1340-1345. https://doi.org/10.1016/j.asr.2005.10.007.

33. Krisanova N.V., Trikash I.O., Borisova T.A. Synaptopathy under conditions of altered gravity: changes in synaptic vesicle fusion and glutamate release. Neurochem Int. 2009, 55 (8), 724-731. https://doi.org/10.1016/J.NEUINT.2009.07.003.

34. Larson E., Howlett B., Jagendorf A. Artificial reductant enhancement of the Lowry method for protein determination. Anal Biochem. 1986, 155 (2), 243-248. https://doi.org/10.1016/0003-2697(86)90432-X.

35. Pozdnyakova N., Pastukhov A., Dudarenko M., Galkin M., Borysov A., Borisova T. Neuroactivity of detonation nanodiamonds: dose- dependent changes in transporter-mediated uptake and ambient level of excitatory/inhibitory neurotransmitters in brain nerve terminals. J Nanobiotechnology. 2016, 14 (1), 25. https://doi.org/10.1186/s12951-016-0176-y.

36. Krisanova N., Pozdnyakova N., Pastukhov A., Dudarenko M., Maksymchuk O, Parkhomets P., Sivko R., Borisova T. Vitamin D3 deficiency in puberty rats causes presynaptic malfunctioning through alterations in exocytotic release and uptake of glutamate/ GABA and expression of EAAC-1/GAT-3 transporters. Food Chem Toxicol. 2019, 123, 142-150. https://doi.org/10.1016/j.fct.2018.10.054.

37. Borisova T., Krisanova N., Sivko R., Borysov A. Cholesterol depletion attenuates tonic release but increases the ambient level of glutamate in rat brain synaptosomes. Neurochem Int. 2010, 56 (3), 466-478. https://doi.org/10.1016/j.neuint.2009.12.006.

38. Borisova T., Borysov A. Putative duality of presynaptic events. Rev Neurosci. 2016, 27 (4), 377-83. https://doi.org/10.1515/revneuro-2015-0044.

39. Borisova T., Borysov A., Pastukhov A., Krisanova N. Dynamic Gradient of Glutamate Across the Membrane: Glutamate/Aspartate-Induced Changes in the Ambient Level of L-[(14)C]glutamate and D-[(3)H]aspartate in Rat Brain Nerve Terminals. Cell Mol Neurobiol. 2016, 36 (8), 1229-1240. https://doi.org/10.1007/s10571-015-0321-4.

40. Borisova T., Nazarova A., Dekaliuk M., Krisanova N., Pozdnyakova N., Borysov A., Sivko R., Demchenko AP. Neuromodulatory properties of fluorescent carbon dots: Effect on exocytotic release, uptake and ambient level of glutamate and GABA in brain nerve terminals. Int J Biochem Cell Biol. 2015, 59, 203-15. https://doi.org/10.1016/j.biocel.2014.11.016.

41. Krisanova., Dudarenko M.V., Pastukhov A.O., Sivko R.V., Kalynovska L.M., Driuk M.M., Nazarova A.G., Gutich І.І., Shliakhovyi V.V., Pozdnyakova N.G. Evaluation of the potential neuroactivity in the brain nerve terminals of the C60 fullerene planetary dust component. Sp Sci Technol. 2023, 29 (5). https://doi.org/10.15407/knit2023.05.060.

42. Pastukhov A., Paliienko K., Pozdnyakova N., Krisanova N., Dudarenko M., Kalynovska L., Tarasenko A., Gnatyuk O., Dovbeshko G., Borisova T. Disposable facemask waste combustion emits neuroactive smoke particulate matter. Sci Rep. 2023, 13 (1), 17771. https:// doi.org/10.1038/s41598-023-44972-0.

43. Madadrang C.J., Kim H.Y., Gao G., Wang N., Zhu J., Feng H., Gorring M., Kasner M.L., Hou S. Adsorption behavior of EDTA-graphene oxide for Pb (II) removal. ACS Appl.

44. Mater Interfaces. 2012, 4 (3), 1186-1193. https://doi.org/10.1021/AM201645G.

45. Mi X., Huang G., Xie W., Wang W., Liu Y., Gao J. Preparation of graphene oxide aerogel and its adsorption for Cu 2+ ions. Carbon N Y. 2012, 50 (13), 4856-4864. https://doi.org/10.1016/J.CARB0N.2012.06.013.

46. Deng X., Lu L, Li H., Luo F. The adsorption properties of Pb(II) and Cd(II) on functionalized graphene prepared by electrolysis method. J Hazard Mater. 2010, 183 (1-3), 923-930. https://doi.org/10.1016/J. JHAZMAT.2010.07.117.

47. Zhao G., Jiang L., He Y., Li J., Dong H., Wang X., Hu W. Sulfonated graphene for persistent aromatic pollutant management. Adv Mater. 2011, 23 (34), 3959-3963. https://doi.org/10.1002/ADMA.201101007.

48. Zhao G., Li J., Ren X., Chen C., Wang X. Few-layered graphene oxide nanosheets as superior sorbents for heavy metal ion pollution management. Environ Sci Technol. 2011, 45 (24), 10454-10462. https://doi.org/10.1021/ES203439V.

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