The Effect of Pristine and Pegylated Graphene Oxide Nanosheets on the Functions of Human Neutrophils
Abstract
Graphene oxide (GO) is very useful for biomedicine, due to its physicochemical properties; therefore, its interaction with cells of the immune system has beenextensively studied. Many studies have aimed toreduce the undesirable effects of GO through chemical modification, including through polyethylene glycol (PEG) coating. Neutrophils are the first to respond to foreign object invasion in the body. Their main functions are the uptake and destruction of foreign particles, including with the help of reactive oxygen species (ROS).Our study aimed to investigate theengulfment of unmodified graphene oxide (GO) and graphene oxide coated with polyethylene glycol (GO-PEG) by human neutrophils and the effect of nanosheets on the production of ROS.We used sheets of GO (Ossila, Great Britain, average plate size 1-5 μm) and GO-PEG (569 ± 14 nm, PEG coating≈ 20%) at concentrations of 12.5μg/mL, 25μg/mL, and 50 μg/mL. The uptake of nanosheets was assessed by flow cytometry, taking into account the level of background adhesion of nanoparticles. ROS production was evaluated by luminol-dependent chemiluminescence (LCL).It was found that GO (12.5μg/mL, 25μg/mL, and 50 μg/mL) was actively internalized by neutrophils, while the uptake of GO-PEG was not detected. GO and GO-PEG particles (25 μg/mLand 50 μg/mL) reduced the total production of ROS by human leukocytes.Thus, the modifying of GOnanosheets with PEG resulted in the abolishment of their active uptake by neutrophils but did not affect the GO inhibitory effect on their oxidative activity.
Keywords: graphene oxide surface modification, pegylated graphene oxide nanosheets, nanoparticle uptake, human neutrophils, of reactive oxygen species
References
[1] Razumov VF.Graphene is a new breakthrough in nanotechnology. Russian Nanotechnology. 2010;5:17-22.
[2] Pankratov D.V., Gonsales-Arribas E., Parunova Y.M. et al. New nanobiocomposite materials for bioelectronic devices. Acta Naturae. 2015;7:103-107.
[3] Shareena D.T.P., Mcshan D., Dasmahapatra A., Tchounwou P. A review on graphenebased nanomaterials in biomedical applications and risks in environment and health. Nano-Micro Letters.2018; 10(53), 1-34 .
[4] Zhang H, Yan T., Xu S. et al. Graphene oxide-chitosan nanocomposites for intracellular delivery of immunostimulatory CpG oligodeoxynucleotides. Materials Science & Engineering C-Materials for Biological Applications. 2017; 1(73),144-151.
[5] Bi S, Zhao T,Luo B. A graphene oxide platform for the assay of biomolecules based on chemiluminescence resonance energy transfer. Chemical Communications. 2012;48(1). 106-108.
[6] Tabish TA, Zhang S, Winyard PG. Developing the next generation of graphene-based platforms for cancer therapeutics: The potential role of reactive oxygen species. Redox Biology. 2018;15. 34-40
[7] Kurapati R, Russier J, Squillaci MA et al. Dispersibility-dependent biodegradation of graphene oxide by myeloperoxidase. Small. 2016;11(32). 3985-3994
[8] Paino IM, Santos F, Zucolotto V. Biocompatibility and toxicology effects of graphene oxide in cancer, normal, and primary immune cells. Journal of Biomedical Materials Research - Part A. 2017;105(3). 728-736
[9] Mendes RG, Mandarino A., Koch B. et al. Size and time dependent internalization of label-free nano-graphene oxide in human macrophages. Nano Research. 2017;10. 1980–1995
[10] Mukherjee SP, Bottini M, Fadeel B. Graphene and the immune system: A romance of many dimensions. Frontiers in Immunology. 2017;8. 673-674
[11] Hermanson GT. Bioconjugate chemistry. 2nd edition. San Diego: Academic Press;2008.
[12] Zamorina SA, Timganova V., Bochkova M., Khramtsov P. Effect of graphene oxide nanoparticles on the differentiation of myeloid suppressor cells.Byulleten’ Eksperimental’noybiologiiimeditsiny. 2020;170(7):102-105.
[13] Santos JL, Montes MJ, Gutiérrez F, Ruiz C. Evaluation of phagocytic capacity with a modified flow cytometry technique. Immunology Letters. 1995;45. 1-4.
[14] Russier J, Treossi E, Scarsi A et al. Evidencing the mask effect of graphene oxide: A comparative study on primary human and murine phagocytic cells. Nanoscale. 2013;5. 11234-11247
[15] Sanchez L,Yi Y, YuY. Effect of partial PEGylation on particle uptake by macrophages. Nanoscale. 2017;91, 288-297 doi.org/10.1039/C6AN00325G
[16] Bisso PW, Gaglione S, Guimarães PPG, Mitchell MJ, Langer R Nanomaterial interactions with human neutrophils. ACS Biomaterials Science and Engineering. 2018;4(12). 4255-4265 doi.org/410.1021/acsbiomaterials.8b01062
[17] Baali N., Khecha A., Bensouici A., Speranza G., Hamdouni N. Assessment of antioxidant activity of pure graphene oxide (GO) and ZnO-decorated reduced graphene Oxide (rGO) using DPPH radical and H2O2 scavenging assays. Journal of Carbon Research. 2019;5. 752-759 doi.org/10.3390/c5040075
[18] Markovic ZM, Jovanovic SP, Maškovic PZ et al. Graphene oxide size and structure pro-oxidant and antioxidant activity and photoinduced cytotoxicity relation on three cancer cell lines. Journal of Photochemistry and PhotobiologyB: Biology. 2019;200. 111647-111648
[19] Qiu Y, Wang Z, Owens AC et al. Antioxidant chemistry of graphene-based materials and its role in oxidation protection technology. Nanoscale. 2014;6(20). 11744-11755
[20] Han J, Kim YS, Lim MY et al. Dual roles of graphene oxide to attenuate inflammation and elicit timely polarization of macrophage phenotypes for cardiac repair. ACS Nano. 2018;12 (2) 1959-1977