Immunotoxicity of stainless-steel nanoparticles obtained after 3D printing.

This study aims to investigate the in vitro effects of nanoparticles (NPs) produced during the selective laser melting (SLM) of 316 L stainless steel metal powder on the immune response in a human blood model. Experimental data did not reveal effect on viability of 316 L NPs for the tested doses. Functional immune assays showed a significant immunosuppressive effect of NPs. There was moderate stimulation (117%) of monocyte phagocytic activity without significant changes in phagocytic activity and respiratory burst of granulocytes. A significant dose-dependent increase in the levels of the pro-inflammatory cytokine TNF-a was found in blood cultures treated with NPs. On the contrary, IL-8 chemokine levels were significantly suppressed. The levels of the pro-inflammatory cytokine IL-6 were reduced by only a single concentration of NPs. These new findings can minimise potential health risks and indicate the need for more research in this area.

Simple method for quantification of metal-based particles in biopsy samples of patients with long bone implants - Pilot study.

The presence of particles fixed in tissue samples due to implant degradation or disintegration plays an important role in post-operative complications. The ability to determine the size, shape, chemical composition and, above all, the number of these particles can be used in many areas of medicine. This study presents a novel, simple metal-based particle detection method using scanning electron microscopy with energy dispersive spectrometer (SEM-EDS). The presence of metal particles in biopsy specimens from long bone nail-fixated implants (10 patients with titanium steel nails and 10 patients with stainless steel nails) was studied. The samples were analysed using automated area analysis based on image binarization and brightness to 255 grayscale. The results were supplemented with histological data and statistically analysed. The method based on the software used was found to be accurate and easy to use and, thus, appears to be very suitable for particle detection in similar samples.

Detection of Micron and Submicron Particles in Human Bronchogenic Carcinomas.

Metal based particles were detected in pulmonary tumor tissue samples and reference lung tissue samples (lung tissue without carcinoma) by Raman microspectroscopy and scanning electron microscopy. Many of these particles were in the size below 1 µm. Using scanning electron microscopy, particles based on iron were found in the majority of samples. Siderite was determined as the form of the iron in several samples by Raman microspectroscopy. The hypothesis that significant statistical dependence exists between the presence of metals in the lung tissue and lung cancer incidence was not proved. However, statistical dependence between smoking and lung cancer incidence was determined as well as insignificant correlation between the presence of titanium based particles and lung cancer incidence. Titanium-based compounds were identified as TiO2 in the form of anatase and rutile. Both the reference and the carcinoma samples exhibited magnetic properties as confirmed by vibration magnetometry measurements.

Settled iron-based road dust and its characteristics and possible association with detection in human tissues.

Settled road dust was examined to detect the presence of non-airborne submicron and nano-sized iron-based particles and to characterize these particles. Samples were collected from a road surface near a busy road junction in the city of Ostrava, Czech Republic, once a month from March to October. The eight collected samples were subjected to a combination of experimental techniques including elemental analysis, Raman microspectroscopy, scanning electron microscopy (SEM) analysis, and magnetometry. The data thereby obtained confirmed the presence of non-agglomerated spherical nano-sized iron-based particles, with average sizes ranging from 2 down to 490 nm. There are several sources in road traffic which generate road dust particles, including exhaust and non-exhaust processes. Some of them (e.g., brake wear) produce iron as the dominant metallic element. Raman microspectroscopy revealed forms of iron (mainly as oxides, Fe2O3, and mixtures of Fe2O3 and Fe3O4). Moreover, Fe3O4 was also detected in samples of human tissues from the upper and lower respiratory tract. In view of the fact that no agglomeration of those particles was found by SEM, it is supposed that these particles may be easily resuspended and represent a risk to human health due to inhalation exposure, as proved by the detection of particles with similar morphology and phase composition in human tissues.

Biological response of an in vitro human 3D lung cell model exposed to brake wear debris varies based on brake pad formulation.

Wear particles from automotive friction brake pads of various sizes, morphology, and chemical composition are significant contributors towards particulate matter. Knowledge concerning the potential adverse effects following inhalation exposure to brake wear debris is limited. Our aim was, therefore, to generate brake wear particles released from commercial low-metallic and non-asbestos organic automotive brake pads used in mid-size passenger cars by a full-scale brake dynamometer with an environmental chamber simulating urban driving and to deduce their potential hazard in vitro. The collected fractions were analysed using scanning electron microscopy via energy-dispersive X-ray spectroscopy (SEM-EDS) and Raman microspectroscopy. The biological impact of the samples was investigated using a human 3D multicellular model consisting of human epithelial cells (A549) and human primary immune cells (macrophages and dendritic cells) mimicking the human epithelial tissue barrier. The viability, morphology, oxidative stress, and (pro-)inflammatory response of the cells were assessed following 24 h exposure to ~ 12, ~ 24, and ~ 48 µg/cm2 of non-airborne samples and to ~ 3.7 µg/cm2 of different brake wear size fractions (2-4, 1-2, and 0.25-1 µm) applying a pseudo-air-liquid interface approach. Brake wear debris with low-metallic formula does not induce any adverse biological effects to the in vitro lung multicellular model. Brake wear particles from non-asbestos organic formulated pads, however, induced increased (pro-)inflammatory mediator release from the same in vitro system. The latter finding can be attributed to the different particle compositions, specifically the presence of anatase.

Toxicity and mutagenicity of low-metallic automotive brake pad materials.

Organic friction materials are standardly used in brakes of small planes, railroad vehicles, trucks and passenger cars. The growing transportation sector requires a better understanding of the negative impact related to the release of potentially hazardous materials into the environment. This includes brakes which can release enormous quantities of wear particulates. This paper addresses in vitro detection of toxic and mutagenic potency of one model and two commercially available low-metallic automotive brake pads used in passenger cars sold in the EU market. The model pad made in the laboratory was also subjected to a standardized brake dynamometer test and the generated non-airborne wear particles were also investigated. Qualitative "organic composition" was determined by GC/MS screening of dichloromethane extracts. Acute toxicity and mutagenicity of four investigated sample types were assessed in vitro by bioluminescence assay using marine bacteria Vibrio fischeri and by two bacterial bioassays i) Ames test on Salmonella typhimurium His(-) and ii) SOS Chromotest using Escherichia coli PQ37 strain. Screening of organic composition revealed a high variety of organic compounds present in the initial brake pads and also in the generated non-airborne wear debris. Several detected compounds are classified by IARC as possibly carcinogenic to humans, e. g. benzene derivatives. Acute toxicity bioassay revealed a response of bacterial cells after exposure to all samples used. Phenolic resin and wear debris were found to be acutely toxic; however in term of mutagenicity the response was negative. All non-friction exposed brake pad samples (a model pad and two commercial pad samples) were mutagenic with metabolic activation in vitro.

Oxidative stress markers are elevated in exhaled breath condensate of workers exposed to nanoparticles during iron oxide pigment production.

Markers of oxidative stress and inflammation were analysed in the exhaled breath condensate (EBC) and urine samples of 14 workers (mean age 43 ± 7 years) exposed to iron oxide aerosol for an average of 10 ± 4 years and 14 controls (mean age 39 ± 4 years) by liquid chromatography-electrospray ionization-mass spectrometry/mass spectrometry (LC-ESI-MS/MS) after solid-phase extraction. Aerosol exposure in the workplace was measured by particle size spectrometers, a scanning mobility particle sizer (SMPS) and an aerodynamic particle sizer (APS), and by aerosol concentration monitors, P-TRAK and DustTRAK DRX. Total aerosol concentrations in workplace locations varied greatly in both time and space. The median mass concentration was 0.083 mg m(-3) (IQR 0.063-0.133 mg m(-3)) and the median particle concentration was 66 800 particles cm(-3) (IQR 16,900-86,900 particles cm(-3)). In addition, more than 80% of particles were smaller than 100 nm in diameter. Markers of oxidative stress, malondialdehyde (MDA), 4-hydroxy-trans-hexenale (HHE), 4-hydroxy-trans-nonenale (HNE), 8-isoProstaglandin F2α (8-isoprostane) and aldehydes C6-C12, in addition to markers of nucleic acid oxidation, including 8-hydroxy-2-deoxyguanosine (8-OHdG), 8-hydroxyguanosine (8-OHG), 5-hydroxymethyl uracil (5-OHMeU), and of proteins, such as o-tyrosine (o-Tyr), 3-chlorotyrosine (3-ClTyr), and 3-nitrotyrosine (3-NOTyr) were analysed in EBC and urine by LC-ESI-MS/MS. Almost all markers of lipid, nucleic acid and protein oxidation were elevated in the EBC of workers comparing with control subjects. Elevated markers were MDA, HNE, HHE, C6-C10, 8-isoprostane, 8-OHdG, 8-OHG, 5-OHMeU, 3-ClTyr, 3-NOTyr, o-Tyr (all p < 0.001), and C11 (p < 0.05). Only aldehyde C12 and the pH of samples did not differ between groups. Markers in urine were not elevated. These findings suggest the adverse effects of nano iron oxide aerosol exposure and support the utility of oxidative stress biomarkers in EBC. The analysis of urine oxidative stress biomarkers does not support the presence of systemic oxidative stress in iron oxide pigment production workers.

Raman microspectroscopy of exhaled breath condensate and urine in workers exposed to fine and nano TiO2 particles: a cross-sectional study.

The health effects of engineered nanoparticles in humans are not well-understood; however experimental data support the theory of oxidative stress promoting fibrogenesis and carcinogenicity. The aim of this study was to detect TiO2 particles in exhaled breath condensate (EBC) and urine samples to ascertain their presence and potential persistence and excretion in urine.EBC and urine samples were collected from 20 workers exposed to TiO2 aerosol; among them, 16 had a higher risk level of exposure (production workers) and four had medium risk level (research workers); in addition to 20 controls. Titanium levels in EBC and urine were analysed using the inductively coupled plasma mass spectrometry (ICP-MS) method. A Raman microspectroscopic analysis was performed in EBC and urine to identify the phase composition of TiO2 particles observed. Aerosol exposure in the workplaces was measured using SMPS and APS spectrometers and P-TRAK and DustTRAK DRX monitors.The median concentration of TiO2 aerosol was 1.98 × 10(4) particles cm(-3), the interquartile range (IQR) was 1.50 × 10(4) - 3.01 × 10(4) particles cm(-3) and the median mass concentration was 0.65 mg m(-3) (IQR 0.46-.0.83 mg m(-3)); 70-82% of the particles were smaller than 100 nm in diameter. In any part of the plant, the median TiO2 air concentration did not exceed the national airborne exposure limit of 10 mg m(-3) for inert dust. Particles of rutile and/or anatase were found in the EBC of exposed workers in 8/20 (40%) of the pre-shift and 14/20 (70%) of the post-shift samples. In the urine of workers, TiO2 particles were detected in 2/20 post-shift urine samples only. The mean concentration of titanium in the EBC in production workers was 24.1 ± 1.8 µg/l. In the research workers the values were below the limit of quantitation; LOQ = 4.0 ± 0.2 µg/l), as well as in the controls. In the urine samples of all of the subjects, titanium was under the limit of detection (LOD = 1.2 µg/l). Raman microanalysis of EBC in the workers confirmed the presence of TiO2 anatase and/or rutile crystal phases in the pre-shift samples and their persistence from previous shifts in the workers.

Iron-based granules in body of bumblebees.

The paper deals with the presence of iron-based granules in body parts of bumblebees. Two groups of bumblebees were collected from their natural habitat, industrial landscape, and from a breeding station. Detection of the magnetic particles was performed by a vibratory magnetometer and their morphology and elemental composition was analysed by scanning electron microscopy with EDX microanalysis. By means of the EDX spectra, wild bumblebees were found to have many magnetic and non-magnetic particles on their body, containing Fe, O, Al, Si, Bi, Mg, K, and Ni, likely having origin in the industrial pollution of the environment. In the case of bred bumblebees the presence of iron-rich granules, which occurred more abundantly in subsurface tissues on the head and wings, was observed. Phase analysis based on X-ray diffraction shows that iron-based granules contain magnetite and wuestite and Mössbauer spectroscopy admits a superparamagnetic form of these minerals. Magnetoreception, i.e. the sensory function of these granules, is discussed within the paper.

Detection of nano- and micro-sized particles in routine biopsy material - pilot study.

BACKGROUND: Nanotechnology is receiving enormous funding. Very little however is known about the health dangers of this technology so far. Chronic tonsillitis is one of a number of diseases called idiopathic. Among other factors, the tonsils are exposed to suspended particles in inhaled air including nano particles. The objective of this study was to detect and evaluate metallic particles in human tonsil tissue diagnosed with chronic tonsillitis and in amniotic fluid as a comparison. METHODS: . Scanning electron microscopy with energy dispersive X-ray spectroscopy (SEM-EDX) was used for identification of solid particles in a total of 64 samples of routinely analyzed biopsy and cytologic material. RESULTS: Almost all samples were found to contain solid particles of various metals. The most frequent, regardless of diagnosis, were iron, chromium, nickel and aluminium. The size, determined using SEM, varied from around 500 nm to 25 µm. The majority formed aggregates of several micrometers in size but there were a significant number of smaller (sub-micrometer or nano-sized) particles present. The incidence of metallic particles was similar in child and adult tissues. The difference was in composition: the presence of several metals in adults was due to occupational exposure. CONCLUSIONS: The presence of metallic particles in pathologically altered tissues may signal an alternative causation of some diseases. The ethiopathogenic explanation of these diseases associated with the presence of nano-sized particles in the organism has emerged into a new field of pathology, nanopathology.

Possible role of nano-sized particles in chronic tonsillitis and tonsillar carcinoma: a pilot study.

This study aimed to evaluate the palatine tonsils of patients with chronic tonsillitis and spinocellular carcinoma to determine the presence of nano-sized particles. Tonsil samples from adult patients with chronic tonsillitis and spinocellular carcinoma of the palatine tonsil were dried and analyzed using a scanning electron microscope with the X-ray microprobe of an energy-dispersive spectroscope. Demographic data and smoking histories were obtained. The principal metals found in almost all tissues analyzed were iron, chromium, nickel, aluminum, zinc, and copper. No significant difference in elemental composition was found between the group of patients with chronic tonsillitis and the group with spinocellular carcinoma of the palatine tonsil. Likewise, no significant difference was found between the group of smokers and the group of nonsmokers. The presence of various micro- and nano-sized metallic particles in human tonsils was confirmed. These particles may potentially cause an inflammatory response as well as neoplastic changes in human palatine tonsils similar to those occurring in the lungs. Further and more detailed studies addressing this issue, including studies designed to determine the chemical form of the metals detected, studies devoted to quantitative analysis, biokinetics, and to the degradation and elimination of nanoparticles are needed for a more detailed prediction of the relation between the diagnosis and the presence of specific metal nanoparticles in tonsillar tissue.

On airborne nano/micro-sized wear particles released from low-metallic automotive brakes.

The paper addresses the wear particles released from commercially available "low-metallic" automotive brake pads subjected to brake dynamometer tests. Particle size distribution was measured in situ and the generated particles were collected. The collected fractions and the original bulk material were analyzed using several chemical and microscopic techniques. The experiments demonstrated that airborne wear particles with sizes between 10 nm and 20 µm were released into the air. The numbers of nanoparticles (< 100 nm) were by three orders of magnitude larger when compared to the microparticles. A significant release of nanoparticles was measured when the average temperature of the rotor reached 300°C, the combustion initiation temperature of organics present in brakes. In contrast to particle size distribution data, the microscopic analysis revealed the presence of nanoparticles, mostly in the form of agglomerates, in all captured fractions. The majority of elements present in the bulk material were also detected in the ultra-fine fraction of the wear particles.