The Relevance of Physico-Chemical Properties and Protein Corona for Evaluation of Nanoparticles Immunotoxicity-In Vitro Correlation Analysis on THP-1 Macrophages.

Alongside physiochemical properties (PCP), it has been suggested that the protein corona of nanoparticles (NPs) plays a crucial role in the response of immune cells to NPs. However, due to the great variety of NPs, target cells, and exposure protocols, there is still no clear relationship between PCP, protein corona composition, and the immunotoxicity of NPs. In this study, we correlated PCP and the protein corona composition of NPs to the THP-1 macrophage response, focusing on selected toxicological endpoints: cell viability, reactive oxygen species (ROS), and cytokine secretion. We analyzed seven commonly used engineered NPs (SiO2, silver, and TiO2) and magnetic NPs. We show that with the exception of silver NPs, all of the tested TiO2 types and SiO2 exhibited moderate toxicities and a transient inflammatory response that was observed as an increase in ROS, IL-8, and/or IL-1ß cytokine secretion. We observed a strong correlation between the size of the NPs in media and IL-1ß secretion. The induction of IL-1ß secretion was completely blunted in NLR family pyrin domain containing 3 (NLRP3) knockout THP-1 cells, indicating activation of the inflammasome. The correlations analysis also implicated the association of specific NP corona proteins with the induction of cytokine secretion. This study provides new insights toward a better understanding of the relationships between PCP, protein corona, and the inflammatory response of macrophages for different engineered NPs, to which we are exposed on a daily basis.

Enhanced osteogenesis on titanium implants by UVB photofunctionalization of hydrothermally grown TiO2 coatings.

Even though Ti-based implants are the most used materials for hard tissue replacement, they may present lack of osseointegration on the long term, due to their inertness. Hydrothermal treatment (HT) is a useful technique for the synthesis of firmly attached, highly crystalline coatings made of anatase titanium dioxide (TiO2), providing favorable nanoroughness and higher exposed surface area, as well as greater hydrophilicity, compared to the native amorphous oxide on pristine titanium. The hydrophilicity drops even more by photofunctionalization of the nanostructured TiO2-anatase coatings under UV light. Human mesenchymal stem cells exhibited a good response to the combination of the positive surface characteristics, especially in respect to the UVB pre-irradiation. The results showed that the cells were not harmed in terms of viability; even more, they were encouraged to differentiate in osteoblasts and to become osteogenically active, as confirmed by the calcium ion uptake and the formation of well-mineralized, bone-like nodule structures. In addition, the enrichment of hydroxyl groups on the HT-surfaces by UVB photofunctionalization accelerated the cell differentiation process and greatly improved the osteogenesis in comparison with the nonirradiated samples. The optimal surface characteristics of the HT-anatase coatings as well as the high potentiality of the photo-induced hydrophilicity, which was reached during a relatively short pre-irradiation time (5 h) with UVB light, can be correlated with better osseointegration ability in vivo; among the samples, the superior biological behavior of the roughest and most hydrophilic HT coating makes it a good candidate for further studies and applications.

Pre-irradiation of anatase TiO2 particles with UV enhances their cytotoxic and genotoxic potential in human hepatoma HepG2 cells.

Titanium dioxide (TiO(2)) is active in the UV region of the light spectra and is used as a photocatalyst in numerous applications. Photo-activated anatase TiO(2) particles promote increased production of free radicals. This is a desirable property, although the potential toxicity of such photo-activated TiO(2) particles on exposure of humans and the environment remains unknown. Therefore, we studied whether pre-irradiation of TiO(2) particles with UV influences their cytotoxic and genotoxic potential. The TiO(2) particles, as TiO(2)-A (<25 nm) and TiO(2)-B (>100 nm), were UV pre-irradiated (24h) and tested for cytotoxic and genotoxic activities in human hepatoma HepG2 cells. Non-irradiated TiO(2)-A/B at 1.0-250 µg/ml did not reduce viability of HepG2 cells, nor induce significant increases in DNA strand breaks; only TiO(2)-A induced significant increases in oxidative DNA damage. After UV pre-irradiation, both TiO(2)-A and TiO(2)-B reduced cell viability and induced significant increases in DNA strand breaks and oxidative DNA damage. This is the first study that shows that UV pre-irradiation of anatase TiO(2) particles results in increased cytotoxic and genotoxic potential. This warrants further studies as it has important implications for environmental and human health risk assessment and preventive actions to limit human exposure.

DNA damage and alterations in expression of DNA damage responsive genes induced by TiO2 nanoparticles in human hepatoma HepG2 cells.

We investigated the genotoxic responses to two types of TiO2 nanoparticles (<25 nm anatase: TiO(2)-An, and <100 nm rutile: TiO2-Ru) in human hepatoma HepG2 cells. Under the applied exposure conditions the particles were agglomerated or aggregated with the size of agglomerates and aggregates in the micrometer range, and were not cytotoxic. TiO2-An, but not TiO2-Ru, caused a persistent increase in DNA strand breaks (comet assay) and oxidized purines (Fpg-comet). TiO2-An was a stronger inducer of intracellular reactive oxygen species (ROS) than TiO2-Ru. Both types of TiO2 nanoparticles transiently upregulated mRNA expression of p53 and its downstream regulated DNA damage responsive genes (mdm2, gadd45α, p21), providing additional evidence that TiO2 nanoparticles are genotoxic. The observed differences in responses of HepG2 cells to exposure to anatase and rutile TiO2 nanoparticles support the evidence that the toxic potential of TiO2 nanoparticles varies not only with particle size but also with crystalline structure.

Titanium dioxide in our everyday life; is it safe?

BACKGROUND: Titanium dioxide (TiO(2)) is considered as an inert and safe material and has been used in many applications for decades. However, with the development of nanotechnologies TiO(2) nanoparticles, with numerous novel and useful properties, are increasingly manufactured and used. Therefore increased human and environmental exposure can be expected, which has put TiO(2) nanoparticles under toxicological scrutiny. Mechanistic toxicological studies show that TiO(2) nanoparticles predominantly cause adverse effects via induction of oxidative stress resulting in cell damage, genotoxicity, inflammation, immune response etc. The extent and type of damage strongly depends on physical and chemical characteristics of TiO(2) nanoparticles, which govern their bioavailability and reactivity. Based on the experimental evidence from animal inhalation studies TiO(2) nanoparticles are classified as "possible carcinogenic to humans" by the International Agency for Research on Cancer and as occupational carcinogen by the National Institute for Occupational Safety and Health. The studies on dermal exposure to TiO(2) nanoparticles, which is in humans substantial through the use of sunscreens, generally indicate negligible transdermal penetration; however data are needed on long-term exposure and potential adverse effects of photo-oxidation products. Although TiO(2) is permitted as an additive (E171) in food and pharmaceutical products we do not have reliable data on its absorption, distribution, excretion and toxicity on oral exposure. TiO(2) may also enter environment, and while it exerts low acute toxicity to aquatic organisms, upon long-term exposure it induces a range of sub-lethal effects. CONCLUSIONS: Until relevant toxicological and human exposure data that would enable reliable risk assessment are obtained, TiO(2) nanoparticles should be used with great care.

[Topographic localization of an intraocular foreign body by B-scan echography].

BACKGROUND/AIM: In cases of blurred optic media the ultrasound diagnostics offers useful data about eventual presence of intraocular foreign body as well as about its precise localization in the eye. The aim of this study was to retrospectively analyze echographic findings in patients with the diagnosis of intraocular foreign body with a special interest in localizations of a intraocular foreign body in the eye and the presence of an eventual infection - endophthalmitis. The aim of this study was also to confirm the localization of intraocular foreign body by echography and to test the precision of this method. METHODS: We performed analysis of all cases that had been referred to the ultrasound diagnostices, in which the presence of intraocular foreign body had been confirmed in the period of one year. All examinations were done with B-scan and were confirmed during the surgery - vitrectomy. RESULTS: In the one-year period we were contacted by 27 patients with intraocular foreign body. In one injured eye the intraocular foreign body was in the lens (3.70%), in 10 injured eyes (37.03%) intraocular foreign body was in the vitreal body. In 15 patients (55.5%) intraocular foreign body was fixed in the retina. In one patient (3.70%) there was a perforating injury, intraocular foreign body was found in the retrobulbar part of the orbit. In 7 injured eyes (25.9%), with the presence of intraocular foreign body, we found signs of endophthalmitis (organized blurring in vitreal space, thickened choroid). Other accompanying echographic findings were: blood in vitreal space, haemophthalmus in 12 cases (44.4%), retinal detachment in 5 cases (18.5%) and subretinal hemorrhagies in 4 cases (14.8%). CONCLUSION: Ultrasound diagnostics can very precisely show the localization of intraocular foreign body in the eye that is very important in the choice of approach and timing of surgical treatment. Also the echographic diagnostics may find an accompanying endophthalmitis in the posterior segment of the eye, that is very important for an urgent therapeutic approach.