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.

Alteration of Hordeum vulgare and Sinapis alba germination and early growth in response to airborne low-metallic automotive brake wear debris.

Road transportation significantly contributes to environmental pollution, both in terms of exhaust and non-exhaust (brake wear) emissions. As was proven, brake wear debris is released in a wide variety of sizes, shapes, and compositions. Although studies confirming the possible adverse health and environmental impact of brake wear debris were published, there is no standardized methodology for their toxicity testing, and most studies focus only on one type of brake pad and/or one test. The lack of methodology is also related to the very small amount of material released during the laboratory testing. For these reasons, this study deals with the mixture of airborne brake wear debris from several commonly used low-metallic brake pads collected following the dynamometer testing. The mixture was chosen for better simulation of the actual state in the environment and to collect a sufficient amount of particles for thorough characterization (SEM, XRPD, XRF, chromatography, and particle size distribution) and phytotoxicity testing. The particle size distribution measurement revealed a wide range of particle sizes from nanometers to hundreds of nanometers, elemental and phase analysis determined the standard elements and compounds used in the brake pad formulation. The Hordeum vulgare and Sinapis alba were chosen as representatives of monocotyledonous and dicotyledonous plants. The germination was not significantly affected by the suspension of brake wear debris; however, the root elongation was negatively influenced in both cases. Sinapis alba (IC50 = 23.13 g L-1) was more affected than Hordeum vulgare (IC50 was not found in the studied concentration range) the growth of which was even slightly stimulated in the lowest concentrations of brake wear debris. The plant biomass was also negatively affected in the case of Sinapis alba, where the IC50 values of wet and dry roots were determined to be 44.83 g L-1 and 86.86 g L-1, respectively.

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.

Microplastics and additives in patients with preterm birth: The first evidence of their presence in both human amniotic fluid and placenta.

Considering the well documented, almost ubiquitous nature of microplastics in different environments, the exposure of pregnant women to microplastics may pose risks to the unborn children. The study focused on investigating the presence of microplastics in amniotic fluid and placenta, and brings the first evidence of the simultaneous presence of microplastics and additives in both human amniotic fluid and placentas. In total, 20 samples of amniotic fluid and placenta from 10 patients were analyzed for the presence of microplastics and plastic additives by Fourier transform infrared spectroscopy - attenuated total reflectance (FTIR-ATR) after alkaline digestion with KOH. In 9 out of 10 patients, microplastics or additives were found in amniotic fluid, placenta, or both. Specifically, 44 particles of microplastics and polymer additives were identified in all samples. Chlorinated Polyethylene (CPE) and Calcium zinc PVC Stabilizer with particle sizes between 10 and 50 µm prevailed. Although all women involved in this study, who provided placenta and amniotic fluid samples, experienced physiological, singleton pregnancies complicated with preterm prelabour rupture of membranes (PPROM), it is too early to draw any conclusions and more research is needed.

Solid Anorganic Particles and Chronic Rhinosinusitis: A Histopathology Study.

Although extensive research has shown the pathological effect of fine and ultrafine airborne particles, clear evidence of association of environmental exposure to them and inflammatory changes in human nasal mucosa is missing. Meanwhile, pathogenesis of chronic rhinosinusitis, despite being a disease with high prevalence in the population, is still unclear. The increasing evidence of the pro-inflammatory properties of these particles raises the question of their possible role in chronic rhinosinusitis. The presented study focused on detection of microsized anorganic particles and clusters of nanosized anorganic particles in the nasal mucosa of patients with chronic rhinosinusitis by Raman microspectroscopy and comparison of their composition to histologic findings. The results were compared to the findings in mucosa obtained from cadavers with no history of chronic rhinosinusitis. Solid particles were found in 90% of tissue samples in the group with chronic rhinosinusitis, showing histologic signs of inflammation in 95%, while in the control group, the particles were found in 20% of samples, with normal histologic findings in all of them. The main detected compounds were graphite, TiO2, amorphous carbon, calcite, ankerite and iron compounds. The results are in accordance with the premise that exogenous airborne particles interact with the nasal mucosa and possibly deposit in it in cases where the epithelial barrier is compromised in chronic rhinosinusitis.

Traffic pollution tracers in the lymphatic system tissue of children-possible link to chronic tonsillitis development: pilot study.

The origin of metal elements in tonsillar tissues of children patients was investigated in order to establish the link between environmental pollution and tonsilitis diagnosis. The children lived either in a polluted, industrial region (Moravian-Silesian Region, North-East Czechia) or a clean, predominantly agricultural region (South Bohemian Region, South-East Czechia). Simultaneously, the distance of the address of the patients to the closest main road was assessed. The elemental content of the present particulate matter in the tonsil samples, coming from indicated tonsillectomies, was analysed using scanning electron microscopy equipped with an X-ray microprobe. No significant association of the elemental composition and the region was found using both bivariate and multivariate methods; however, an association with the distance to the closest main road was found. The study presents a novel assessment of the tonsillar tissues and supports the link between environmental factors (traffic-related particulate pollution) and the onset of chronic, idiopathic diseases in children, which has been debated recently.

Identification of the phase composition of solid microparticles in the nasal mucosa of patients with chronic hypertrophic rhinitis using Raman microspectroscopy.

Solid particles, predominantly in micron and submicron sizes, have repeatedly been observed as a threat to a human health unique compared to the other textures of the same materials. In this work, the hypothesis the solid metal-based particles play a role in the pathogenesis of chronic hypertrophic rhinitis was investigated in patients who had not responded positively to medication. In the group of 40 randomly selected patients indicated for surgical mucotomy, the presence of solid micro- and submicron particles present in their nasal mucosa was assessed. For comparison, a set of 13 reference samples from patients without diagnosed chronic hypertrophic rhinitis was evaluated. The analysis was performed using Raman microspectroscopy. The advantage of this method is the direct identification of compounds. The main detected compounds in the mucosa samples of patients with chronic hypertrophic rhinitis were TiO2, carbon-based compounds, CaCO3, Ca(Fe, Mg, Mn)(CO3)2 MgCO3, Fe2O3, BaSO4, FeCO3 and compounds of Al and Si, all of which may pose a health risk to a living organism. In the reference samples, only TiO2 and amorphous carbon were found. In the control group mucosa, a significantly lower presence of most of the assessed compounds was found despite the longer time they had to accumulate them due to their higher mean age. Identification and characterisation of such chemicals compounds in a living organism could contribute to the overall picture of the health of the individual and lead to a better understanding of the possible causes not only in the chronic hypertrophic rhinitis, but also in other mucosal and idiopathic diseases.

Size-resolved, quantitative evaluation of the magnetic mineralogy of airborne brake-wear particulate emissions.

Exposure to particulate air pollution has been associated with a variety of respiratory, cardiovascular and neurological problems, resulting in increased morbidity and mortality worldwide. Brake-wear emissions are one of the major sources of metal-rich airborne particulate pollution in roadside environments. Of potentially bioreactive metals, Fe (especially in its ferrous form, Fe2+) might play a specific role in both neurological and cardiovascular impairments. Here, we collected brake-wear particulate emissions using a full-scale brake dynamometer, and used a combination of magnetic measurements and electron microscopy to make quantitative evaluation of the magnetic composition and particle size of airborne emissions originating from passenger car brake systems. Our results show that the concentrations of Fe-rich magnetic grains in airborne brake-wear emissions are very high (i.e., ~100-10,000 × higher), compared to other types of particulate pollutants produced in most urban environments. From magnetic component analysis, the average magnetite mass concentration in total PM10 of brake emissions is ~20.2 wt% and metallic Fe ~1.6 wt%. Most brake-wear airborne particles (>99 % of particle number concentration) are smaller than 200 nm. Using low-temperature magnetic measurements, we observed a strong superparamagnetic signal (indicative of ultrafine magnetic particles, < ~30 nm) for all of the analysed size fractions of airborne brake-wear particles. Transmission electron microscopy independently shows that even the larger size fractions of airborne brake-wear emissions dominantly comprise agglomerates of ultrafine (<100 nm) particles (UFPs). Such UFPs likely pose a threat to neuronal and cardiovascular health after inhalation and/or ingestion. The observed abundance of ultrafine magnetite particles (estimated to constitute ~7.6 wt% of PM0.2) might be especially hazardous to the brain, contributing both to microglial inflammatory action and excess generation of reactive oxygen species.

Source apportionment of magnetite particles in roadside airborne particulate matter.

Exposure to airborne particulate matter (PM) is associated with pulmonary, cardiovascular and neurological problems. Magnetite, a mixed Fe2+/Fe3+ oxide, is ubiquitous and abundant in PM in urban environments, and might play a specific role in both neurodegeneration and cardiovascular disease. We collected samples of vehicle exhaust emissions, and of heavily-trafficked roadside and urban background dusts from Lancaster and Birmingham, U.K. Then, we measured their saturation magnetic remanence and used magnetic component analysis to separate the magnetite signal from other contributing magnetic components. Lastly, we estimated the contributions made by specific traffic-related sources of magnetite to the total airborne magnetite in the roadside environment. The concentration of magnetite in exhaust emissions is much lower (3-14 x lower) than that in heavily- trafficked roadside PM. The magnetite concentration in petrol-engine exhaust emissions is between ~0.06 and 0.12 wt%; in diesel-engine exhaust emissions ~0.08-0.18 wt%; in background dust ~0.05-0.20 wt% and in roadside dust ~0.18-0.95 wt%. Here, we show that vehicle brake wear is responsible for between ~68 and 85% of the total airborne magnetite at the two U.K. roadside sites. In comparison, diesel-engine exhaust emissions account for ~7% - 12%, petrol-engine exhaust emissions for ~2% - 4%, and background dust for 6% - 10%. Thus, vehicle brake wear is by far the most dominant source of airborne magnetite in the roadside environment at the two sites examined. Given the potential risk posed, post-inhalation, by ultrafine magnetite and co-associated transition metal-rich particles to human cardiovascular and neurological health, the high magnetite content of vehicle brake wear might need to be reduced in order to mitigate such risk, especially for vulnerable population groups.

Metal particles in mucus and hypertrophic tissue of the inferior nasal turbinates from the human upper respiratory tract.

Mucosal surfaces are the first mechanical barrier preventing the entry of foreign particles into the organism. The study addresses the detection and analysis of metal-based solid particles in cytological mucus samples from the surface of human hypertrophic tissue in the inferior nasal turbinates in patients diagnosed with chronic rhinitis. Solid particles were characterized by scanning electron microscopy and Raman microspectroscopy; all the biological samples were also subjected to vibration magnetometry. Since the upper airways are the first part of the respiratory tract, which is exposed to inhaled particles, it can be assumed that inhaled particles may be partially deposited in this region. Scanning electron microscopy revealed the presence of metal-based solid particles/clusters in the majority of the analysed cytological mucus samples and also in hypertrophic tissues; in all groups, the particles were of submicron size. Raman microspectroscopy detected the presence of particles/clusters based on amorphous carbon, graphite, calcium carbonate, anatase and barite only in the hypertrophic tissue. The obtained results show that the composition of some of the solid particles (i.e. Ba, Zn, Fe and Ti) detected in the mucus from the surface of the hypertrophic tissues resembled the particles found in the hypertrophic tissue itself. It can be assumed that after the capture of the inhaled particles by the mucus, they penetrate into the deeper layers of tissue.

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.

Determination of Oxidative Potential Caused by Brake Wear Debris in Non-Cellular Systems.

Wear debris from automotive brake systems represents a major source of non-exhaust emissions from road traffic and its production increases with number of cars worldwide. However, impact of brake wear debris on the environment and organisms is still not clear. One of the most possible ways by which these particles may affect living organisms is oxidative stress. Production of reactive oxidative species may cause damage of basic cell components, lipids, proteins, etc. Aim of this study is to perform characterization of airborne and nonairborne fractions of brake wear debris generated during standard dynamometer tests and evaluation of its potential to induce oxidative stress via lipid peroxidation and carbonylation of proteins in non-cellular system. Elemental and phase composition were determined by scanning electron microscopy, Raman microspectroscopy, and X-ray powder diffraction analysis. Carbon in amorphous form and graphite, copper, and iron in form of oxides were identified as major components in both studied fractions. Characteristic size of studied wear particles was evaluated by dynamic light scattering. Both airborne and nonairborne samples showed ability to induce oxidative stress which results from determination of carbonylated proteins.

Synthesis and Characterization of Erbium Oxide Nanocrystallites.

The present article describes a method of the preparation of erbium oxide nanocrystallites (nano Er2O3) via thermal decomposition of a transient complex formed in situ from Er(NO3)3·5H2O and glycine. Decomposition of the complex occurred at about of (250±10) °C. Ultra-fine light pink powder of erbium oxide nanocrystallites was obtained via this method. The resulting nanocrystallites were characterized using X-ray powder diffraction analysis, which showed the nanocrystallites having the crystallite size equal to 10 nm. Morphology of the nanocrystallites was examined by scanning and transmission electron microscopy. Electron diffraction observed in transmission electron microscopy corresponds to the results obtained from X-ray diffraction analysis. The elemental composition of the product was confirmed by EDS analysis.

Settled Dust from Urban and Suburban Roads in an Industrial City Area: Location and Seasonal Differences in Metal Content.

Settled road dust, present on all roads surfaces in a relatively high abundance, is a complex of particles-ranging from nanosized to microsized-from both natural and anthropogenic sources and may pose possible risk to the biosphere as well as influence the atmosphere because of the road dust resuspension. Geographical and seasonal differences in heavy metal content of the settled road dust were studied at two sites: urban site with a heavy traffic in the industrial city of Ostrava, Czech Republic and suburban site of Ostrava with a negligible traffic load in settlement Ludgerovice. Dust samples were collected monthly during the period from March to October 2015. Obtained dust samples were homogenized and subsequently analyzed by Inductively Coupled Plasma Atomic Emission Spectroscopy; the results were then assessed by multivariate statistical methods (Principal components analysis, Factorial analysis on mixed data). The difference in the content of Ba, Cr, Cu, Mn, Ni, Pb, Sb, Sn, V, and Zn was explainable mainly by the factor of the site only the content of Fe was significantly dependent on the month of sampling. The contents of the particular elements and the correlations found among them confirm the assumption that heavy metals detected in the road dust samples from the urban site most likely originated primarily from the traffic-and particularly from the non-combustion processes.

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.

Automotive airborne brake wear debris nanoparticles and cytokinesis-block micronucleus assay in peripheral blood lymphocytes: A pilot study.

Motor vehicle exhaust and non-exhaust processes play a significant role in environmental pollution, as they are a source of the finest particulate matter. Emissions from non-exhaust processes include wear-products of brakes, tires, automotive hardware, road surface, and traffic signs, but still are paid little attention to. Automotive friction composites for brake pads are composite materials which may consist of potentially hazardous materials and there is a lack of information regarding the potential influence of the brake wear debris (BWD) on the environment, especially on human health. Thus, we focused our study on the genotoxicity of the airborne fraction of BWD using a brake pad model representing an average low-metallic formulation available in the EU market. BWD was generated in the laboratory by a full-scale brake dynamometer and characterized by Raman microspectroscopy, scanning electron microscopy, and transmission electron microscopy showing that it contains nano-sized crystalline metal-based particles. Genotoxicity tested in human lymphocytes in different testing conditions showed an increase in frequencies of micronucleated binucleated cells (MNBNCs) exposed for 48h to BWD nanoparticles (NPs) (with 10% of foetal calf serum in culture medium) compared with lymphocytes exposed to medium alone, statistically significant only at the concentration 3µg/cm(2) (p=0.032).

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.

ZnO/graphite composites and its antibacterial activity at different conditions.

The paper reports laboratory preparation, characterization and in vitro evaluation of antibacterial activity of ZnO/graphite nanocomposites. Zinc chloride and sodium carbonate served as precursors for synthesis of zinc oxide, while micromilled and natural graphite were used as the matrix for ZnO nanoparticles anchoring. During the reaction of ZnCl2 with saturated aqueous solution of Na2CO3a new compound is created. During the calcination at the temperature of 500 °C this new precursors decomposes and ZnO nanoparticles are formed. Composites ZnO/graphite with 50 wt.% of ZnO particles were prepared. X-ray powder diffraction and Raman microspectroscopy served as phase-analytical methods. Scanning electron microscopy technique was used for morphology characterization of the prepared samples and EDS mapping for visualization of elemental distribution. A developed modification of the standard microdilution test was used for in vitro evaluation of daylight induced antibacterial activity and antibacterial activity at dark conditions. Common human pathogens served as microorganism for antibacterial assay. Antibacterial activity of ZnO/graphite composites could be based on photocatalytic reaction; however there is a role of Zn(2+) ions on the resulting antibacterial activity which proved the experiments in dark condition. There is synergistic effect between Zn(2+) caused and reactive oxygen species caused antibacterial activity.