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Article

Dispersion and Dosimetric Challenges of Hydrophobic Carbon-Based Nanoparticles in In Vitro Cellular Studies

by
Denisa Lizonova
1,
Una Trivanovic
2,
Philip Demokritou
1,* and
Georgios A. Kelesidis
1,2,*
1
Nanoscience and Advanced Materials Center (NAMC), Environmental and Occupational Health Science Institute, School of Public Health, Rutgers University, 170 Frelinghuysen Road, Piscataway, NJ 08854, USA
2
Particle Technology Laboratory, Institute of Energy and Process Engineering, Department of Mechanical and Process Engineering, ETH Zürich, Sonneggstrasse 3, CH-8092 Zürich, Switzerland
*
Authors to whom correspondence should be addressed.
Nanomaterials 2024, 14(7), 589; https://doi.org/10.3390/nano14070589
Submission received: 31 January 2024 / Revised: 18 March 2024 / Accepted: 21 March 2024 / Published: 27 March 2024

Abstract

Methodologies across the dispersion preparation, characterization, and cellular dosimetry of hydrophilic nanoparticles (NPs) have been developed and used extensively in the field of nanotoxicology. However, hydrophobic NPs pose a challenge for dispersion in aqueous culture media using conventional methods that include sonication followed by mixing in the culture medium of interest and cellular dosimetry. In this study, a robust methodology for the preparation of stable dispersions of hydrophobic NPs for cellular studies is developed by introducing continuous energy over time via stirring in the culture medium followed by dispersion characterization and cellular dosimetry. The stirring energy and the presence of proteins in the culture medium result in the formation of a protein corona around the NPs, stabilizing their dispersion, which can be used for in vitro cellular studies. The identification of the optimal stirring time is crucial for achieving dispersion and stability. This is assessed through a comprehensive stability testing protocol employing dynamic light scattering to evaluate the particle size distribution stability and polydispersity. Additionally, the effective density of the NPs is obtained for the stable NP dispersions using the volumetric centrifugation method, while cellular dosimetry calculations are done using available cellular computational modeling, mirroring approaches used for hydrophilic NPs. The robustness of the proposed dispersion approach is showcased using a highly hydrophobic NP model (black carbon NPs) and two culture media, RPMI medium and SABM, that are widely used in cellular studies. The proposed approach for the dispersion of hydrophobic NPs results in stable dispersions in both culture media used here. The NP effective density of 1.03–1.07 g/cm3 measured here for black carbon NPs is close to the culture media density, resulting in slow deposition on the cells over time. So, the present methodology for dispersion and dosimetry of hydrophobic NPs is essential for the design of dose–response studies and overcoming the challenges imposed by slow particle deposition.
Keywords: hydrophobic nanoparticles; black carbon; dispersion; protein stabilization; cell culture; in vitro testing; effective density; dosimetry hydrophobic nanoparticles; black carbon; dispersion; protein stabilization; cell culture; in vitro testing; effective density; dosimetry

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MDPI and ACS Style

Lizonova, D.; Trivanovic, U.; Demokritou, P.; Kelesidis, G.A. Dispersion and Dosimetric Challenges of Hydrophobic Carbon-Based Nanoparticles in In Vitro Cellular Studies. Nanomaterials 2024, 14, 589. https://doi.org/10.3390/nano14070589

AMA Style

Lizonova D, Trivanovic U, Demokritou P, Kelesidis GA. Dispersion and Dosimetric Challenges of Hydrophobic Carbon-Based Nanoparticles in In Vitro Cellular Studies. Nanomaterials. 2024; 14(7):589. https://doi.org/10.3390/nano14070589

Chicago/Turabian Style

Lizonova, Denisa, Una Trivanovic, Philip Demokritou, and Georgios A. Kelesidis. 2024. "Dispersion and Dosimetric Challenges of Hydrophobic Carbon-Based Nanoparticles in In Vitro Cellular Studies" Nanomaterials 14, no. 7: 589. https://doi.org/10.3390/nano14070589

APA Style

Lizonova, D., Trivanovic, U., Demokritou, P., & Kelesidis, G. A. (2024). Dispersion and Dosimetric Challenges of Hydrophobic Carbon-Based Nanoparticles in In Vitro Cellular Studies. Nanomaterials, 14(7), 589. https://doi.org/10.3390/nano14070589

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