RESUMEN
Although several studies assess the biological effects of micro and titanium dioxide nanoparticles (TiO2 NPs), the literature shows controversial results regarding their effect on bone cell behavior. Studies on the effects of nanoparticles on mammalian cells on two-dimensional (2D) cell cultures display several disadvantages, such as changes in cell morphology, function, and metabolism and fewer cell-cell contacts. This highlights the need to explore the effects of TiO2 NPs in more complex 3D environments, to better mimic the bone microenvironment. This study aims to compare the differentiation and mineralized matrix production of human osteoblasts SAOS-2 in a monolayer or 3D models after exposure to different concentrations of TiO2 NPs. Nanoparticles were characterized, and their internalization and effects on the SAOS-2 monolayer and 3D spheroid cells were evaluated with morphological analysis. The mineralization of human osteoblasts upon exposure to TiO2 NPs was evaluated by alizarin red staining, demonstrating a dose-dependent increase in mineralized matrix in human primary osteoblasts and SAOS-2 both in the monolayer and 3D models. Furthermore, our results reveal that, after high exposure to TiO2 NPs, the dose-dependent increase in the bone mineralized matrix in the 3D cells model is higher than in the 2D culture, showing a promising model to test the effect on bone osteointegration.
RESUMEN
Titanium dioxide (TiO2) is manufactured worldwide as crystalline and amorphous forms for multiple applications, including tissue engineering, but our study proposes analyzing the impact of crystalline phases of TiO2 on Mesenchymal Stem Cells (MSCs). Several studies have already described the regenerative potential of MSCs and TiO2 has been used for bone regeneration. In this study, polydispersity index and sizes of TiO2 nanocrystals (NCs) were determined. Adipose tissue-derived Mesenchymal Stem Cells (AT-MSCs) were isolated and characterized in order to evaluate cellular viability and the internalization of nanocrystals (NCs). All of the assays were performed using the TiO2 NCs with 100% anatase (A), 91.6% anatase/9.4% rutile (AR), 64.6% rutile/35.4% anatase (RA), and 84.0% rutile/16% brookite (RB), submitted to several concentrations in 24-h treatments. Cellular localization of TiO2 NCs in the AT-MSCs was resolved by europium-doped NCs. Viability was significantly improved under the predominance of the rutile phase in NCs with localization restricted at the cytoplasm, suggesting that AR and RA NCs are not genotoxic and can be associated with most cellular activities and metabolic pathways, including glycolysis and cell division.
RESUMEN
Titanium dioxide nanoparticles (TiO2 NPs) are regularly used in sunscreens because of their photoprotective capacity. The advantage of using TiO2 on the nanometer scale is due to its transparency and better UV blocking efficiency. Due to the greater surface area/volume ratio, NPs become more (bio)-reactive giving rise to concerns about their potential toxicity. To evaluate the irritation and corrosion of cosmetics, 3D skin models have been used as an alternative method to animal experimentation. However, it is not known if this model is appropriate to study skin irritation, corrosion and phototoxicity of nanomaterials such as TiO2 NPs. This systematic review (SR) proposed the following question: Can the toxicity of TiO2 nanoparticles be evaluated in a 3D skin model? This SR was conducted according to the Preliminary Report on Systematic Review and Meta-Analysis (PRISMA). The protocol was registered in CAMARADES and the ToxRTool evaluation was performed in order to increase the quality and transparency of this search. In this SR, 7 articles were selected, and it was concluded that the 3D skin model has shown to be promising to evaluate the toxicity of TiO2 NPs. However, most studies have used biological assays that have already been described as interfering with these NPs, demonstrating that misinterpretations can be obtained. This review will focus in the possible efforts that should be done in order to avoid interference of NPs with biological assays applied in 3D in vitro culture.