Comparative study of the functional activity of blood mononuclear cells by cytokine production under the influence of titanium dioxide nanoparticles with different sulfur content in vitro
DOI:
https://doi.org/10.14739/2310-1237.2025.3.331274Keywords:
anatase, titanium dioxide nanoparticles, sulfur modification, nanomaterials, peripheral blood mononuclear cells, cytokine production, immunotoxicity, in vitro study, in vitro, ELISA, immune responseAbstract
Aim. To investigate the effect of titanium dioxide (TiO2) nanoparticles (anatase form) with varying sulfur content on the functional activity of peripheral blood mononuclear cells (PBMCs) from healthy donors in vitro, based on the production of cytokines IL-1β, IL-4, IL-6, and TNF-α.
Materials and methods. The objects of the study were titanium dioxide (TiO2) nanopowders of the anatase crystalline form, with a particle size of 21-28 nm and varying sulfur content (0.04 %, 0.16 % and 0.83 %), developed at the I. M. Frantsevich Institute for Problems of Materials Science. Peripheral blood mononuclear cells (PBMCs) from healthy volunteer donors (n = 30) were incubated in vitro either without a stimulating agent, with the mitogen phytohemagglutinin (PHA), or in the presence of TiO2 nanoparticle suspensions (10 μL) at concentrations of 0.3 mg/mL, 3 mg/mL, and 30 mg/mL. Cytokine concentrations (IL-1β, IL-6, IL-4, and TNF-α) in the PBMC supernatants were measured using enzyme-linked immunosorbent assay (ELISA). The tests were performed using a Stat Fax-303 Plus ELISA reader.
Results. In vitro studies demonstrated that the nanomaterial at concentrations starting from 0.3 µg/mL, specifically TiO2 nanoparticles with 0.16 % and 0.83 % sulfur content, suppressed the functional activity of peripheral blood mononuclear cells, as evidenced by a statistically significant decrease in the production of cytokines IL-1β, IL-6, TNF-α, and IL-4 in donor samples (p < 0.05). This suggests a potential disruption of immune system function. In contrast, exposure to TiO2 nanoparticles containing 0.04 % sulfur at a concentration of 30 µg/mL resulted in a statistically significant reduction only in TNF-α production (p < 0.05), while IL-6, IL-1β, and IL-4 levels did not significantly differ from spontaneous cytokine production. Moreover, the data indicate that the immunomodulatory effect of TiO2 nanoparticles is closely related to their sulfur content: the higher the sulfur concentration, the more pronounced the suppression of key cytokine production (IL-1β, IL-6, TNF-α, IL-4). Overall, these results highlight the ability of sulfur-modified nanoparticles to exert differential effects on the immune response, ranging from neutral to potentially immunotoxic outcomes.
Conclusions. The functional activity of peripheral blood mononuclear cells, in terms of cytokine production (IL-1β, IL-6, TNF-α, IL-4), is suppressed by the tested TiO2 nanoparticles at specific concentrations, following the order: nano-TiO2 (0.04 % S) < nano-TiO2 (0.83 % S) < nano-TiO2 (0.16 % S). A sulfur content of 0.16 % or higher in TiO2 nanoparticles is associated with increased immunotoxicity. These findings suggest that the effect of TiO2 nanoparticles on cytokine production by peripheral blood mononuclear cells varies depending on the sulfur content, indicating a direct relationship between the chemical composition of the nanomaterial and its potential immunotoxicity.
References
Forest V, Pourchez J, Pélissier C, Audignon Durand S, Vergnon JM, Fontana L. Relationship between occupational exposure to airborne nanoparticles, nanoparticle lung burden and lung diseases. Toxics. 2021;9(9):204. doi: https://doi.org/10.3390/toxics9090204
Simova Z, Sima M, Pelclova D, Klusackova P, Zdimal V, Schwarz J, et al. Transcriptomics insight into occupational exposure to engineered nanoparticles. Nanomedicine (Lond). 2025;20(14):1713-27. doi: https://doi.org/10.1080/17435889.2025.2527020
Romanko M, Orobchenko O, Palliy A, Ushkalov V, Palii A, Chechui H. Evaluation of biochemical markers in the blood plasma of rats exposed to chronic administration of a mixture of metal nanoparticles. Veterinarska Stanica. 2023;54(1):69-85. doi: https://doi.org/10.46419/vs.54.1.10
Barik P, Mondal S. Immunomodulatory effects of metal nanoparticles: current trends and future prospects. Nanoscale. 2025;17(17):10433-61. doi: https://doi.org/10.1039/d5nr01030f
Aljabali AA, Obeid MA, Bashatwah RM, Serrano-Aroca Á, Mishra V, Mishra Y, et al. Nanomaterials and their impact on the immune system. Int J Mol Sci. 2023;24(3):2008. doi: https://doi.org/10.3390/ijms24032008
Barbir R, Capjak I, Crnković T, Debeljak Ž, Domazet Jurašin D, Ćurlin M, et al. Interaction of silver nanoparticles with plasma transport proteins: A systematic study on impacts of particle size, shape and surface functionalization. Chem Biol Interact. 2021;335:109364. doi: https://doi.org/10.1016/j.cbi.2020.109364
Park SJ. Protein–nanoparticle interaction: corona formation and conformational changes in proteins on nanoparticles. Int J Nanomedicine. 2020;15:5783-802. doi: https://doi.org/10.2147/IJN.S254808
Ray P, Haideri N, Haque I, Mohammed O, Chakraborty S, Banerjee S, et al. The impact of nanoparticles on the immune system: a gray zone of nanomedicine. Int J Nanomedicine. 2021;5(1):19-33. doi: https://doi.org/10.29245/2578-3009/2021/1.1206
Olšovská E, Mikušová M, Tulinska J, Rollerova E, Vilamová Z, Liskova A, et al. Immunotoxicity of stainless-steel nanoparticles obtained after 3D printing. Ecotoxicol Environ Saf. 2024;272:116088. doi: https://doi.org/10.1016/j.ecoenv.2024.116088
Vijayaraj S, Feltham R, Rashidi M, Frank D, Liu Z, Simpson D, et al. The ubiquitylation of IL-1β limits its cleavage by caspase-1 and targets it for proteasomal degradation. Nat Commun. 2021;12:22979. doi: https://doi.org/10.1038/s41467-021-22979-3
Said EA, Al-Reesi I, Al-Shizawi N, Jaju S, Al-Balushi MS, Koh CY, et al. Defining IL-6 levels in healthy individuals: A meta-analysis. J Med Virol. 2021;93(6):3915-24. doi: https://doi.org/10.1002/jmv.26654
Lu M, Lee Y, Lillehoj H. Unveiling the immunological attributes of chicken tumor necrosis factor-α (TNF-α) using new sets of monoclonal antibodies specific for poultry TNF-α. J Immunol. 2021. doi: https://doi.org/10.4049/jimmunol.206.supp.19.16
Alathary RA, Abass RA, Al-Muhtaser ST, Al-Fahham AA. Biochemistry, pathophysiology and clinical importance of IL-4: a review article. Int J Health Med Res. 2025;4(1):44-7. doi: https://doi.org/10.58806/ijhmr.2025.v4i1n09
Lehotska Mikusova M, Busova M, Tulinska J, Masanova V, Liskova A, Uhnakova I, et al. Titanium Dioxide Nanoparticles Modulate Systemic Immune Response and Increase Levels of Reduced Glutathione in Mice after Seven-Week Inhalation. Nanomaterials (Basel). 2023;13(4):767. doi: https://doi.org/10.3390/nano13040767
Di Giampaolo L, Zaccariello G, Benedetti A, Vecchiotti G, Caposano F, Sabbioni E, et al. Genotoxicity and Immunotoxicity of Titanium Dioxide-Embedded Mesoporous Silica Nanoparticles (TiO2@MSN) in Primary Peripheral Human Blood Mononuclear Cells (PBMC). Nanomaterials (Basel). 2021;11(2):270. doi: https://doi.org/10.3390/nano11020270
Monopoli MP, Aberg C, Salvati A, Dawson KA. Biomolecular coronas provide the biological identity of nanosized materials. Nat Nanotechnol. 2020;15(7):618-29. doi: https://doi.org/10.1201/9780429399039
Fadeel B, Farcal L, Hardy B, Vázquez-Campos S, Hristozov D, Marcomini A, et al. Advanced tools for the safety assessment of nanomaterials. Nat Nanotechnol. 2022;17(6):611-22. doi: https://doi.org/10.1038/s41565-018-0185-0
Downloads
Additional Files
Published
How to Cite
Issue
Section
License
Copyright (c) 2025 O. P. Yavorovsky, A. I. Kurchenko, V. M. Riabovol, V. S. Savchenko, O. O. Yavorovska
This work is licensed under a Creative Commons Attribution 4.0 International License.
Authors retain copyright and grant the journal right of first publication with the work simultaneously licensed under a Creative Commons Attribution License that allows others to share the work with an acknowledgement of the work's authorship and initial publication in this journal.
