Email this article EVALUATION OF CYTOTOXICITY OF FULLERENOL C60(OH)22-24 TOWARDS HUMAN PERIPHERAL BLOOD NK CELLS IN VITRO
- Authors: Timganova V.P.1, Bochkova M.S.2,3, Usanina D.I.2,3, Dolgikh M.D.2,3, Lazarev S.S.2, Rayev M.B.2,3, Zamorina S.A.2,3
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Affiliations:
- Institute of Ecology and Genetics of Microorganisms of the Ural Branch of the Russian Academy of Sciences"("IEGM Ural Branch of the Russian Academy of Sciences")
- "Institute of Ecology and Genetics of Microorganisms of the Ural Branch of the Russian Academy of Sciences"("IEGM Ural Branch of the Russian Academy of Sciences")
- Perm State University
- Section: Immunological readings in Chelyabinsk
- Submitted: 23.03.2025
- Accepted: 25.05.2025
- URL: https://rusimmun.ru/jour/article/view/17126
- DOI: https://doi.org/10.46235/1028-7221-17126-EOC
- ID: 17126
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Abstract
Abstract
Introduction. Polyhydroxylated fullerenes, commonly referred to as fullerenols, are among the most promising carbon allotropes due to their hydrophilic nature, stability, and low toxicity. Natural Killer (NK) cells, are key components of the antiviral and antitumor immune response. However, they have been largely understudied as targets for fullerenol nanoparticles.
The objective of this study was to assess the immunocompatibility of hydroxylated fullerenol C60(OH)22-24 with СD3-CD56+ NK cells from human peripheral blood, as well as to investigate the internalization of these nanoparticles into cells.
Materials and methods. The studies were conducted on mononuclear cells from the peripheral blood of healthy donors (n=4). In the experiments, fullerenol (MST-WS60-Bio, fullerenol C60(OH)24, 99.99%, Modern Synthesis Technology, Latvia; cluster size approximately 130 nm) was used at concentrations of 200, 100, 50, 25, 12.5, 5, 2.5, 0.5, and 0.25 μg/mL. Wells without added nanoparticles served as controls. Cells were incubated in the presence of fullerenol for 24, 48, and 72 hours under conditions of 5% CO2 and 37°C. The viability of NK cells (CD3-CD56+) and the adhesion/internalization of fullerenol into cells were assessed using flow cytometry.
Results. It was found that fullerenol nanoparticles at concentrations ranging from 0.25 to 200 μg/mL did not exhibit cytotoxicity towards NK cells during the observation periods of 24, 48, and 72 hours. Thus, no statistically significant decrease in the percentage and absolute number of live NK cells was detected in cultures with fullerenol over these time frames. The study also showed that NK cells did not demonstrate adhesion/internalization of fullerenol nanoparticles at low concentrations (0.25-50 μg/mL) during all observation periods. However, high concentrations of fullerenol were detected inside NK cells at 48 and 72 hours of observation. After 72 hours, approximately 20% of NK cells had adhered/internalized fullerenol at a concentration of 100 μg/mL, and about 50% had done so at a concentration of 200 μg/mL.
Conclusion. Thus, for the first time, it was demonstrated that NK cells adhere/internalize fullerenol at high concentrations (100 and 200 μg/mL), and the percentage of fullerenol-positive cells increases with both longer cultivation duration and higher nanoparticle concentration. Fullerenol didn’t exhibit a cytotoxic effect on the studied cell subpopulation.
About the authors
Valeria P. Timganova
Institute of Ecology and Genetics of Microorganisms of the Ural Branch of the Russian Academy of Sciences"("IEGM Ural Branch of the Russian Academy of Sciences")
Email: timganovavp@gmail.com
ORCID iD: 0000-0003-4581-1969
SPIN-code: 9154-2815
PhD (Biology), Research Associate, Laboratory of Cellular Immunology and Nanobiotechnology, Institute of Ecology and Genetic of Microorganisms, Perm Federal Research Center, Ural Branch, Russian Academy of Sciences
Russian Federation, PermMariya S. Bochkova
"Institute of Ecology and Genetics of Microorganisms of the Ural Branch of the Russian Academy of Sciences"("IEGM Ural Branch of the Russian Academy of Sciences");Perm State University
Email: krasnykh-m@mail.ru
ORCID iD: 0000-0001-5784-6224
PhD (Biology), Research Associate, Laboratory of Cell Immunology and Biotechnology
Russian Federation, 13 Golev St., Perm 614081Darya I. Usanina
"Institute of Ecology and Genetics of Microorganisms of the Ural Branch of the Russian Academy of Sciences"("IEGM Ural Branch of the Russian Academy of Sciences");Perm State University
Email: usanina_d@mail.ru
ORCID iD: 0000-0003-0436-0890
SPIN-code: 1225-0890
Junior Research Associate, Laboratory of Molecular Immunology
Russian Federation, 13 Golev St., Perm 614081, Пермь; 15, st. Bukirev, PermMaria Denisovna Dolgikh
"Institute of Ecology and Genetics of Microorganisms of the Ural Branch of the Russian Academy of Sciences"("IEGM Ural Branch of the Russian Academy of Sciences");Perm State University
Email: dolgikhtatanya@gmail.com
ORCID iD: 0009-0002-1418-4157
technician, laboratory of Cellular Immunology and Nanobiotechnology
Russian Federation, 614081, Russia, Perm, Goleva, 13Sergey Stanislavovich Lazarev
"Institute of Ecology and Genetics of Microorganisms of the Ural Branch of the Russian Academy of Sciences"("IEGM Ural Branch of the Russian Academy of Sciences")
Email: lasest1999@gmail.com
ORCID iD: 0000-0002-0229-1812
SPIN-code: 6756-1359
technician, laboratory of Cellular Immunology and Nanobiotechnology
Russian Federation, 614081, Russia, Perm, Goleva, 13Mikhail Borisovich B. Rayev
"Institute of Ecology and Genetics of Microorganisms of the Ural Branch of the Russian Academy of Sciences"("IEGM Ural Branch of the Russian Academy of Sciences");Perm State University
Email: mraev@iegm.ru
ORCID iD: 0000-0001-6882-4928
SPIN-code: 2501-5240
PhD, MD (Biology), Head, Laboratory of Cell Immunology and Biotechnology
Russian Federation, 13 Golev St., Perm 614081Svetlana A. Zamorina
"Institute of Ecology and Genetics of Microorganisms of the Ural Branch of the Russian Academy of Sciences"("IEGM Ural Branch of the Russian Academy of Sciences");Perm State University
Author for correspondence.
Email: mantissa7@mail.ru
ORCID iD: 0000-0002-6474-1487
SPIN-code: 3579-1451
PhD, MD (Biology), Leading Research Associate, Laboratory of Cellular Immunology and Nanobiotechnology, Professor, Department of Microbiology and Immunology
Russian Federation, 13 Golev St., Perm 614081, Пермь; 15, st. Bukirev, PermReferences
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