Effects of physicochemical properties of TiO2 nanomaterials for pulmonary inflammation, acute phase response and alveolar proteinosis in intratracheally exposed mice

Nanomaterial (NM) characteristics may affect the pulmonary toxicity and inflammatory response, including specific surface area, size, shape, crystal phase or other surface characteristics. Grouping of TiO2 in hazard assessment might be challenging because of variation in physicochemical properties. We exposed C57BL/6 J mice to a single dose of four anatase TiO2 NMs with various sizes and shapes by intratracheal instillation and assessed the pulmonary toxicity 1, 3, 28, 90 or 180 days post-exposure. The quartz DQ12 was included as benchmark particle. Pulmonary responses were evaluated by histopathology, electron microscopy, bronchoalveolar lavage (BAL) fluid cell composition and acute phase response. Genotoxicity was evaluated by DNA strand break levels in BAL cells, lung and liver in the comet assay. Multiple regression analyses were applied to identify specific TiO2 NMs properties important for the pulmonary inflammation and acute phase response. The TiO2 NMs induced similar inflammatory responses when surface area was used as dose metrics, although inflammatory and acute phase response was greatest and more persistent for the TiO2 tube. Similar histopathological changes were observed for the TiO2 tube and DQ12 including pulmonary alveolar proteinosis indicating profound effects related to the tube shape. Comparison with previously published data on rutile TiO2 NMs indicated that rutile TiO2 NMs were more inflammogenic in terms of neutrophil influx than anatase TiO2 NMs when normalized to total deposited surface area. Overall, the results suggest that specific surface area, crystal phase and shape of TiO2 NMs are important predictors for the observed pulmonary effects of TiO2 NMs.Peer reviewe

Nanoparticles Can Wrap Epithelial Cell Membranes and Relocate Them Across the Epithelial Cell Layer

Although the link between the inhalation of nanoparticles and cardiovascular disease is well established, the causal pathway between nanoparticle exposure and increased activity of blood coagulation factors remains unexplained. To initiate coagulation tissue factor bearing epithelial cell membranes should be exposed to blood, on the other side of the less than a micrometre thin air-blood barrier. For the inhaled nanoparticles to promote coagulation, they need to bind lung epithelial-cell membrane parts and relocate them into the blood. To assess this hypothesis, we use advanced microscopy and spectroscopy techniques to show that the nanoparticles wrap themselves with epithelial-cell membranes, leading to the membrane's disruption. The membrane-wrapped nanoparticles are then observed to freely diffuse across the damaged epithelial cell layer relocating epithelial cell membrane parts over the epithelial layer. Proteomic analysis of the protein content in the nanoparticles wraps/corona finally reveals the presence of the coagulation-initiating factors, supporting the proposed causal link between the inhalation of nanoparticles and cardiovascular disease.European Commission Horizon 2020Wellcome TrustSlovenian Research AgencyWolfson FoundationOxford-internal fundin

Bacterial adhesion on prosthetic and orthotic material surfaces

Prosthetic and orthotic parts, such as prosthetic socket and inner sides of orthoses, are often in contact with human skin, giving bacteria the capability to adhere and form biofilms on the materials of those parts which can further cause infections. The purpose of this study was to determine the extent of bacterial adhesion of Staphylococcus aureus and Staphylococcus epidermidis on twelve different prosthetic and orthotic material surfaces and how roughness, hydrophobicity, and surface charge of this materials affect the adhesion. The roughness, contact angle, zeta potential of material surfaces, and adhesion rate of Staphylococcus aureus and Staphylococcus epidermidis were measured on all twelve prosthetic and orthotic materials, i.e., poly(methyl methacrylate), thermoplastic elastomer, three types of ethylene polyvinyl acetates (pure, with low-density polyethylene and with silver nanoparticles), silicone, closed-cell polyethylene foams with and without nanoparticles, thermo and natural cork, and artificial and natural leather. The greatest degree of adhesion was measured on both closed-cell polyethylene foams, followed by artificial thermo cork and leather. The lowest adhesion extent was observed on ethylene-vinyl acetate. The bacterial adhesion extent increases with the increasing surface roughness. Smaller deviations of this rule are the result of the surface’s hydrophobicity and charge

A method for targeting a specified segment of DNA to a bacterial microorganelle

Encapsulation of a selected DNA molecule in a cell has important implications for bionanotechnology. Non-viral proteins that can be used as nucleic acid containers include proteinaceous subcellular bacterial microcompartments (MCPs) that self-assemble into a selectively permeable protein shell containing an enzymatic core. Here, we adapted a propanediol utilization (Pdu) MCP into a synthetic protein cage to package a specified DNA segment in vivo, thereby enabling subsequent affinity purification. To this end, we engineered the LacI transcription repressor to be routed, together with target DNA, into the lumen of a Strep-tagged Pdu shell. Sequencing of extracted DNA from the affinity-isolated MCPs shows that our strategy results in packaging of a DNA segment carrying multiple LacI binding sites, but not the flanking regions. Furthermore, we used LacI to drive the encapsulation of a DNA segment containing operators for LacI and for a second transcription factor

Measurements of hydroxyl radical formations by UV irradiation of the TiO₂-NTs.

<p>Spin trap DMPO was used to detect production of the hydroxyl radicals generated by UV irradiated TiO₂-NTs. TiO₂-NTs were mixed with DMPO spin traps and cell medium with 10% FBS or without FBS. The sample was irradiated for 5 min with UV light (wavelength of 356 nm) or left in dark (control), followed by EPR measurements immediately after the addition of the cell medium. In parallel experiments the dispersion of the TiO₂-NTs in the cell medium without or with the serum proteins (FBS) was put in the dark for one hour, than the spin trap DMPO was added and samples were irradiated with the UV light. (A) Representative EPR spectrum of a trapped hydroxyl radical in the presence of the FBS or (B) absence of the FBS. (C) The experimental EPR spectrum was simulated with hyperfine splitting constants: A_N = 1.49 G and A_H = 1.49 mT typical for OH radical. Simulation was done with EPRSim Wizard [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0129577#pone.0129577.ref029" target="_blank">29</a>]. EPR spectrum intensity peak normalized to the experiment with highest intensity peak, of nanomaterial in the cell medium without the FBS and with the FBS.</p

Cell viability versus concentration of TiO₂-NTs in absence or presence of UV radiation.

<p>Absorbance of MTS was measured as described in Materials and methods section. (A) Each non-irradiated data set (red symbols) is compared to a sample irradiated with UV light (blue symbols) at given concentration of TiO₂-NTs. All Measurements are presented in a boxplot representing the distribution of the measured cell viability and described in Materials and methods. Differences in the cell viability between irradiated and non-irradiated cells are shown with the black arrows at each concentration of TiO₂-NTs. The differences in the median values between the two groups at 0 and 1000 μg/mL and the two groups at 1 and 50 μg/mL of nanotubes are considered to be significant at P = <0.001, and P = <0.05, respectively. (B) Differences of median values of MTS absorbance between irradiated and non-irradiated cells, shown with the black arrows within the frame A, are plotted with the open bars. Solid black line represents the prediction of the difference in cell viability, based on the effect of the UV irradiation and phototoxic effect due to irradiated nanotubes not covered by serum proteins. Both contributions are shown separately in the frame C. (C) Solid dark blue line shows the transmittance of UV light versus the concentration of the nanotubes obtained experimentally (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0129577#pone.0129577.s002" target="_blank">S2 Information</a>. Optical properties of TiO₂-NTs dispersion.), while dashed red line represents the concentration of the free nanotubes calculated as described in the <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0129577#pone.0129577.s003" target="_blank">S3 Information</a>. Model of albumin binding to TiO2-NTs and best fit parameter values. (D) The relative amount of TiO₂-NTs covered by the serum proteins (red line) and the relative amount of serum proteins bound to TiO₂-NTs (yellow line) are shown as predicted by the model described in the <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0129577#pone.0129577.s003" target="_blank">S3 Information</a>. Model of albumin binding to TiO2-NTs and best fit parameter values.</p