Functionalized Titanium Nanoparticles Induce Oxidative Stress and Cell Death in Human Skin Cells

Patricia Brassolatti; Joice Margareth de Almeida Rodolpho; Krissia Franco de Godoy; Cynthia Aparecida de Castro; Genoveva Lourdes Flores Luna; Bruna Dias de Lima Fragelli; Matheus Pedrino; Marcelo Assis; Marcel Nani Leite; Juliana Cancino-Bernardi; Carlos Speglich; Marco Andrey Frade; Fernanda de Freitas Anibal

doi:10.2147/IJN.S325767

Functionalized Titanium Nanoparticles Induce Oxidative Stress and Cell Death in Human Skin Cells

Int J Nanomedicine. 2022 Mar 30:17:1495-1509. doi: 10.2147/IJN.S325767. eCollection 2022.

Authors

Affiliations

¹ Laboratory of Inflammation and Infectious Diseases, Department of Morphology and Pathology, Federal University of São Carlos, São Carlos, São Paulo, Brazil.
² Center for the Development of Functional Materials, Department of Chemistry, Federal University of São Carlos, São Carlos, São Paulo, Brazil.
³ Division of Dermatology - Wound Healing & Hansen's Disease Lab, Department of Internal Medicine, Ribeirão Preto Medical School, University of São Paulo, Ribeirão Preto, São Paulo, Brazil.
⁴ Nanomedicine and Nanotoxicology Group, Physics Institute of São Carlos, University of São Paulo, São Carlos, São Paulo, Brazil.
⁵ Leopoldo Américo Miguez de Mello CENPES/Petrobras Research Center, Rio de Janeiro, Rio de Janeiro, Brazil.

Abstract

Purpose: Nanoparticles are resources of advanced nanotechnology being present in several products. Titanium dioxide nanoparticles are among the five most widely used NP currently expanding their benefits from the oil industry to the areas of diagnostic medicine due to their properties and small size. However, its impact on human health is still controversial in the literature. We aimed to evaluate the cytotoxicity of a new titanium NP functionalized with sodium carboxylic ligand (COOH^-Na⁺) in human keratinocytes (HaCaT) and human fibroblasts (HDFn).

Methods: The physical-chemical characterization was performed by the transmission electron microscopy (TEM), dynamic light scattering (DLS) and zeta potential techniques, respectively. MTT and LDH assays were used to assess cytotoxicity and cell membrane damage respectively, ELISA to identify the inflammatory profile and, reactive oxygen species assay and cytometry to detect reactive oxygen species and their relationship with apoptosis/necrosis mechanisms.

Results: The results demonstrated a decrease in cell viability at the highest concentrations tested for both cell lines, but no change in LDH release was detected for the HaCaT. The cell membrane damage was found only at 100.0 µg/mL for the HDFn. It was demonstrated that cytotoxicity in the highest concentrations evaluated for both cell lines for the 72 h period. The HDFn showed damage to the cell membrane at a concentration of 100 µg/mL followed by a significant increase in reactive oxygen species production. No inflammatory profile was detected. The HaCaT showed apoptosis when exposed to the highest concentration evaluated and HDFn showed both apoptosis and necrosis for the same concentration.

Conclusion: Thus, it is possible to conclude that the cytotoxicity mechanism differs according to the cell type evaluated, with HDFn being the most sensitive line in this case, and this mechanism can be defined in a dose and time dependent manner, since the highest concentrations also triggered death cell.

Keywords: cytotoxicity; human skin cells; nanoparticle; titanium.

MeSH terms

Apoptosis
Cell Survival
Humans
Metal Nanoparticles* / chemistry
Metal Nanoparticles* / toxicity
Nanoparticles* / chemistry
Nanoparticles* / toxicity
Necrosis / chemically induced
Oxidative Stress
Reactive Oxygen Species / metabolism
Titanium / chemistry
Titanium / toxicity

Substances

Reactive Oxygen Species
Titanium