TiO2-MWCNT nanohybrid: Cytotoxicity, protein corona formation and cellular internalisation in RTG-2 fish cell line

Gabriela H Da Silva; Lidiane Silva Franqui; Marcelo A De Farias; Vera Lucia S S De Castro; Hugh J Byrne; Diego S T Martinez; Regina T R Monteiro; Alan Casey

doi:10.1016/j.aquatox.2023.106434

TiO_2-MWCNT nanohybrid: Cytotoxicity, protein corona formation and cellular internalisation in RTG-2 fish cell line

Aquat Toxicol. 2023 Apr:257:106434. doi: 10.1016/j.aquatox.2023.106434. Epub 2023 Feb 19.

Authors

Gabriela H Da Silva¹, Lidiane Silva Franqui², Marcelo A De Farias², Vera Lucia S S De Castro³, Hugh J Byrne⁴, Diego S T Martinez⁵, Regina T R Monteiro⁶, Alan Casey⁴

Affiliations

¹ Brazilian Nanotechnology National Laboratory (LNNano), Brazilian Center for Research in Energy and Materials (CNPEM), Campinas, São Paulo, Brazil; Center of Nuclear Energy in Agriculture (CENA), University of São Paulo (USP), Piracicaba, São Paulo, Brazil; Laboratory of Ecotoxicology and Biosafety, EMBRAPA Environment, Jaguariúna, São Paulo, Brazil; FOCAS Research Institute, TU Dublin, City Campus, Camden Row, Dublin 8, Ireland. Electronic address: gabriela.silva@lnnano.cnpem.br.
² Brazilian Nanotechnology National Laboratory (LNNano), Brazilian Center for Research in Energy and Materials (CNPEM), Campinas, São Paulo, Brazil.
³ Laboratory of Ecotoxicology and Biosafety, EMBRAPA Environment, Jaguariúna, São Paulo, Brazil.
⁴ FOCAS Research Institute, TU Dublin, City Campus, Camden Row, Dublin 8, Ireland.
⁵ Brazilian Nanotechnology National Laboratory (LNNano), Brazilian Center for Research in Energy and Materials (CNPEM), Campinas, São Paulo, Brazil; Center of Nuclear Energy in Agriculture (CENA), University of São Paulo (USP), Piracicaba, São Paulo, Brazil.
⁶ Center of Nuclear Energy in Agriculture (CENA), University of São Paulo (USP), Piracicaba, São Paulo, Brazil.

PMID: 36870176
DOI: 10.1016/j.aquatox.2023.106434

Abstract

Titanium dioxide nanoparticles-multiwalled carbon nanotubes (TiO_2-MWCNT) nanohydrid has an enhanced photocatalytic activity across the visible light with promising applications in environmental remediation, solar energy devices and antimicrobial technologies. However, it is necessary to evaluate the toxicological effects of TiO_2-MWCNT towards safe and sustainable development of nanohybrids. In this work, we studied the cytotoxicity, protein corona formation and cellular internalisation of TiO_2-MWCNT on fibroblasts derived from gonadal rainbow trout tissue (RTG-2) for the first time. This nanohydrid did not show any toxicity effect on RTG-2 cells up to 100 mg L^-1 after 24 h of exposure as monitored by alamar blue, neutral red and trypan blue assays (in presence or absence of foetal bovine serum, FBS). Futhermore, cryo-transmission electron microscopy analysis demonstrated that TiO₂ particles is attached on nanotube surface after FBS-protein corona formation in cell culture medium. Raman spectroscopy imaging showed that TiO_2-MWCNT can be internalised by RTG-2 cells. This work is a novel contribution towards better understanding the nanobiointeractions of nanohydrids linked to their in vitro effects on fish cells in aquatic nanoecotoxicology.

Keywords: Ecotoxicity; Nanosafety; Protein corona; Raman.

MeSH terms

Animals
Cell Line
Fishes
Nanoparticles* / toxicity
Nanotubes, Carbon* / chemistry
Nanotubes, Carbon* / toxicity
Protein Corona* / chemistry
Titanium / chemistry
Titanium / toxicity
Water Pollutants, Chemical* / toxicity

Substances

Protein Corona
Nanotubes, Carbon
Water Pollutants, Chemical
Titanium