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Toxic responses in human lung epithelial cells (BEAS-2B) exposed to particulate matter exhaust emissions from gasoline and biogasoline
Závodná, Táňa; Líbalová, Helena; Vrbová, Kristýna; Sikorová, Jitka; Vojtíšek-Lom, M.; Beránek, V.; Pechout, M.; Kléma, J.; Cigánek, M.; Machala, M.; Neča, J.; Rössner ml., Pavel; Topinka, Jan
2021 - English
Motor vehicle emissions substantially contribute to air pollution worldwide and cause serious health problems. While the deleterious effects of diesel exhaust particulate matter (PM) have been widely studied, much less attention is paid to toxicity of PM emitted by gasoline engines although they also produce considerable amount of PM. The primary objective of this research was to assess toxic potencies of exhaust PM released by conventional gasoline engine fueled with neat gasoline (EU) or gasoline-ethanol blend (15% ethanol, v/v, E15). Despite a similar particle mass (mu g PM/kg fuel) produced by both fuels, PM emitted by E15 contained higher amount of harmful polycyclic aromatic hydrocarbons (PAH) as suggested by chemical analysis. To examine the toxicity of organic PM constituents, human lung BEAS-2B cells were exposed for 4h and 24h to a subtoxic dose of E0 and E15 PM organic extracts. We used genome scale transcriptomic analysis to characterize the toxic response and to identify modulated biological process and pathways. Whereas 4h exposure to both PM extracts resulted in modulation of similar genes and pathways related to lipid and steroid metabolism, activation of PPAR alpha, oxidative stress and immune response, 24h exposure was more specific for each extract, although both induced expression of PAH-metabolic enzymes, modulated metabolism of lipids or activated PPAR alpha, E15 additionally deregulated variety of other pathways. Overall, the PM mass produced by both fuels was similar, however, higher PAH content in E15 PM organic extract may have contributed to more extensive toxic response particularly after 24h exposure in BEAS-2B cells.
Keywords:
particulate matter emissions; gasoline; biofuels; toxicity; gene expression profiling
Available at various institutes of the ASCR
Toxic responses in human lung epithelial cells (BEAS-2B) exposed to particulate matter exhaust emissions from gasoline and biogasoline
Motor vehicle emissions substantially contribute to air pollution worldwide and cause serious health problems. While the deleterious effects of diesel exhaust particulate matter (PM) have been widely ...
Application potential of screening in vitro toxicological assays in qualitative risk assessment of nanomaterials
Závodná, Táňa; Topinka, Jan; Danihelka, J.
2020 - English
Undeniable benefits of engineered nanomaterials might be discredited by their potential enhanced or unexpected toxicity arising from nano-specific properties and behavior. An analysis of the applicability of the traditional chemical risk assessment approach in nanomaterials revealed high levels of uncertainty in both hazard characterization and exposure assessment due to the lack of relevant validated methods and reliable data. This indicates the limited capability of the conventional risk assessment approach to ensure the safe use of nanomaterials. Based on the identified uncertainties, the control banding approach was proposed as a suitable tool for preliminary qualitative risk assessment of nanomaterials in occupational settings. Control banding categorizes hazard and exposure into levels referred to as bands. The combination of the hazard and exposure bands results in a risk band determining the necessary degree of control and regulatory measures. To decrease the number of cases where, based on the precautionary principle, unavailable experimental or field data would lead to the assignment to the highest hazard category requiring costly exposure control, screening evaluation of nanomaterial toxicity was proposed as an additional decision criterion. For this purpose, a battery of in vitro toxicological assays enabling screening evaluation of potential toxic effects of NMs was proposed. The assays evaluate endpoints covering basic toxic effects of substances (cytotoxicity, genotoxicity), as well as known nonspecific mechanisms of toxicity typical for nanomaterials (oxidative stress, inflammation). The proposed risk management strategy is intended to assist small and medium-sized enterprises to implement adequate measures to ensure employee safety.
Keywords:
toxicity; tool; industrial poisons
Available at various institutes of the ASCR
Application potential of screening in vitro toxicological assays in qualitative risk assessment of nanomaterials
Undeniable benefits of engineered nanomaterials might be discredited by their potential enhanced or unexpected toxicity arising from nano-specific properties and behavior. An analysis of the ...
Males-females differences in the spectrum of chromosomal aberrations in the group of nanocomposites production workers
Rössnerová, Andrea; Pelcová, D.; Ždímal, Vladimír; Elzeinova, Fatima; Margaryan, Hasmik; Chvojková, Irena; Topinka, Jan; Schwarz, Jaroslav; Ondráček, Jakub; Koštejn, Martin; Komarc, M.; Vlčková, Š.; Fenclová, Z.; Lischková, L.; Dvořáčková, Š.; Rössner ml., Pavel
2020 - English
An increase in the use of nanomaterials (NM) has been witnessed in many areas of human life. Therefore, assessment of genotoxicity of NM and nanoparticles (NP) is one of the main objectives of genetic toxicology. Despite this fact, human cytogenetic studies following the exposure to NP are still rare. Moreover, no relevant information on possible differences in sensitivity to NP related to gender is available.\n\nIn this study we periodically (in September 2016, 2017 and 2018; pre-shift and post-shift each year) analyzed a group of workers (both genders), working long time in nanocomposites research, and matched controls. Aerosol exposure monitoring of particulate matter including nano-sized fractions was carried out during working shift. Micronucleus assay using Human Pan Centromeric probes, was applied to distinguish, besides the frequency of total MN in binucleated cells (BNC), also other types of chromosomal damage (losses and breaks). Moreover, whole-chromosome painting (WCP) for autosome #1 and both gonosomes (X and Y) were applied in third sampling period (2018) with the aim to identify the particular structural and numerical chromosomal aberrations.\n\nObtained results showed: (i) differences in the risk of exposure to NP related to individual working processes (welding, smelting and machining); (ii) differences in chemical composition of nano-fraction; (iii) no effect of chronic exposure of NP (total MN) opposite to significant effect of acute exposure; (iv) gender-related DNA damage differences (females seem to be more sensitive to chromosomal losses). Additional data from WCP suggested increased frequency of numerical aberrations in gonosomes.
Keywords:
DNA damage; gender; chromosomal aberrations; micronuclei; nanoparticles; occupational exposure
Available in a digital repository NRGL
Males-females differences in the spectrum of chromosomal aberrations in the group of nanocomposites production workers
An increase in the use of nanomaterials (NM) has been witnessed in many areas of human life. Therefore, assessment of genotoxicity of NM and nanoparticles (NP) is one of the main objectives of genetic ...
Ultrafine particles and their possible role in etiology and development of neurodegenerative diseases
Topinka, Jan; Závodná, Táňa; Rössnerová, Andrea; Rössner ml., Pavel
2020 - English
Air pollutants have been shown to cause a vast amount of different adverse health effects. These effects include impairment of many respiratory (e.g. asthma, chronic obstructive pulmonary disease) and cardiovascular (ischemic heart disease, infarction, stroke) diseases. However, in recent years, the evidence showing effects beyond the lungs and circulatory system are becoming more evident. Neurological diseases, namely Alzheimer's disease (AD) has shown to be associated with living near traffic. However, reason for this has remained unresolved until today. Our new H2020 project TUBE aims on revealing the mechanisms of action of ultrafine particles involved in neurological diseases. The TUBE consortium includes experts in areas of aerosol technology, emission research, engine and fuel research, human clinical studies, epidemiology, emission inventories, inhalation toxicology, neurotoxicology and disease mechanism studies. This enables research of resolving the effects of nanoparticles from different traffic modes for both air quality and concomitant toxic effect of these air pollutants. We will investigate adverse effects of air pollutants using cell cultures, animal exposures and volunteered human exposures as well as the material from epidemiological cohort study. These are going to be compared according to inflammatory, cytotoxic and genotoxic changes and furthermore beyond the current state of the art to neurotoxic and brain health effects. With this approach, we are aiming to a comprehensive understanding of the adverse brain effects of nanoparticles from traffic.
Keywords:
Alzheimer's disease; inflammation; neurodegenerative diseases; ultrafine particles
Available at various institutes of the ASCR
Ultrafine particles and their possible role in etiology and development of neurodegenerative diseases
Air pollutants have been shown to cause a vast amount of different adverse health effects. These effects include impairment of many respiratory (e.g. asthma, chronic obstructive pulmonary disease) and ...
The role of dispersion medium on nanoparticle aggregation and size in biological systems
Červená, Tereza; Rössnerová, Andrea; Závodná, Táňa; Vrbová, Kristýna; Sikorová, Jitka; Topinka, Jan; Rössner ml., Pavel
2020 - English
The use of nanomaterials (NMs) in different areas has been rising for more than a decade. Along with this growth, there is visible development of different testing tools and approaches for measuring the actual size of nanomaterials in biological systems. Test conditions during in vitro toxicological assays are different from the standard conditions under which nanomaterials are characterized and careful evaluation of results is needed. The unique properties and range variety of NMs require the close look how the NMs behave in different dispersion medium over time. In this study we present the results of five types of well-characterized NMs (TiO2: NM-101 and NM-103, SiO2: NM-200, Ag: NM-300K and NM-302) of specific size and shape. The hydrodynamic size and Zeta potentials in suspensions were measured using a dynamic light scattering technique (DLS) (Zetasizer Nano ZS, Malvern, UK). The DLS method is suitable for spherical particles, nevertheless, all samples were measured in order to obtain a rough insight into agglomerate formation in the medium. NM300, NM302, and NM200 aggregated rapidly in the media, thus the cells would be most likely exposed to settled big aggregates then small clusters or individual particles. More stable NMs (NM100 and NM103) showed slight grow along with cultivation time or concentration corresponding to cluster formation. Cells exposed to those NMs would be in contact with small clusters and aggregates of NMs. Measured zeta potentials fluctuated around the stability limit corresponding to observed aggregation.
Keywords:
nanomaterials; NMs; DLS; aggregation
Available at various institutes of the ASCR
The role of dispersion medium on nanoparticle aggregation and size in biological systems
The use of nanomaterials (NMs) in different areas has been rising for more than a decade. Along with this growth, there is visible development of different testing tools and approaches for measuring ...
Effect of iron oxide nanoparticles with ascorbic acid on neural stem cells
Jiráková, Klára; Moskvin, Maksym; Horák, Daniel; Jendelová, Pavla
2018 - English
Cells labelled with iron oxide nanoparticles (ION) can be tracked by magnetic resonance imaging (MRI) in several applications. However, various studies demonstrated toxicity and oxidative stress induction associated with nanoparticles exposure. We analysed biologic effects after the exposure of two types of iron oxide nanoparticles (with and without an antioxidative agent, an ascorbic acid) on human neural stem cells. The labelled cells in gel phantoms were detected in MRI and they showed decreased relaxation rates in comparison with control. ION slightly decreased cell proliferation in comparison with unlabelled cells, which was dependent on concentration and presence of ascorbic acid. None of the nanoparticle type showed negative effect on cell viability and both demonstrated minor effect on reactive oxygen species (ROS) formation. Unfortunately, ascorbic acid bound to nanoparticles did not show any effect on ROS attenuation. Cells exposed to both types of nanoparticles showed increased positivity for a phosphorylated form of H2AX a marker of double strand breaks. We showed that ION in low concentrations do not affect cell viability, but have negative effect on cells on DNA level. Their potential use for oxidative stress reduction is dependent on the concentration of ascorbic acid bound to the nanoparticles and this should be further increased.
Keywords:
neural stem cells; nanoparticles; oxidative stress; ascorbic acid
Available at various institutes of the ASCR
Effect of iron oxide nanoparticles with ascorbic acid on neural stem cells
Cells labelled with iron oxide nanoparticles (ION) can be tracked by magnetic resonance imaging (MRI) in several applications. However, various studies demonstrated toxicity and oxidative stress ...
GENE EXPRESSION AND IMMUNOLOGICAL RESPONSE IN MICE EXPOSED TO ZnO NANOPARTICLES
Rössner ml., Pavel; Vrbová, Kristýna; Strapáčová, S.; Rössnerová, Andrea; Ambrož, Antonín; Brzicová, Táňa; Líbalová, Helena; Javorková, Eliška; Zajícová, Alena; Holáň, Vladimír; Kulich, P.; Večeřa, Zbyněk; Mikuška, Pavel; Coufalík, Pavel; Křůmal, Kamil; Čapka, Lukáš; Dočekal, Bohumil; Šerý, Omar; Machala, M.; Topinka, Jan
2018 - English
We analyzed gene expression changes in the lungs and the immunological response in splenocytes of mice exposed by inhalation of ZnO nanoparticles - NP. Adult female ICR mice were treated for three days and three months, respectively. Analysis of differential expression in genes involved in oxidative stress was conducted using quantitative RT-PCR. The potential immunotoxic and immunomodulatory effects of ZnO NP were analyzed by phenotyping and cytokine production by splenocytes after three months exposure. Three days exposure resulted in down-regulation of GCLC, GSR, HMOX-1, NQO-1, NF-kB2, PTGS2 and TXNRD1 mRNA expression, three months exposure increased the expression of these genes. Three months exposure caused a significant decrease in the percentage of granulocytes in the spleen cells, and affected the production of IL-10 and IL-6 by lipopolysaccharide-stimulated leukocytes. In summary, our study revealed changes in the expression of genes involved in the oxidative stress response following acute ZnO NP exposure. Subchronic ZnO NP exposure induced immunomodulatory effects in the spleen.
Keywords:
Zinc oxide nanoparticles; inhalation; gene expression; Immune response
Available at various institutes of the ASCR
GENE EXPRESSION AND IMMUNOLOGICAL RESPONSE IN MICE EXPOSED TO ZnO NANOPARTICLES
We analyzed gene expression changes in the lungs and the immunological response in splenocytes of mice exposed by inhalation of ZnO nanoparticles - NP. Adult female ICR mice were treated for three ...
WHOLE-GENOME EXPRESSION ANALYSIS IN THP-1 MACROPHAGE-LIKE CELLS EXPOSED TO DIVERSE NANOMATERIALS
Brzicová, Táňa; Líbalová, Helena; Vrbová, Kristýna; Sikorová, Jitka; Philimonenko, Vlada; Kléma, J.; Topinka, Jan; Rössner ml., Pavel
2018 - English
From the perspective of the immune system, nanomaterials (NMs) represent invading agents. Macrophages are immune cells residing in all organs and tissues as the first line of defense. Interactions of macrophages with NMs can determine the fate of NMs as well as their potential toxic effects. In the present study, we compared toxicity of four different types of NMs [NM-100 (TiO2, 110 nm), NM-110 (ZnO, 20 nm), NM-200 (SiO2, 150 nm) and NM-300K (Ag, 20 nm)], towards THP-1 macrophage-like cells. Cells were incubated with non-cytotoxic concentrations (1-25 mu g/ml) of NMs for 24 hours and microarray technology was used to analyze changes in whole-genome expression. Gene expression profiling revealed a substantially different molecular response following exposure to diverse NMs. While NM-100 did not exert any significant effect on gene expression profile, all other NMs triggered a pro-inflammatory response characterized by an activation of the NF-kappa B transcription factor and induced expression of numerous chemokines and cytokines. NM-110 and NM-300K further modulated processes such as DNA damage response, oxidative and replication stress as well as cell cycle progression and proteasome function. We suppose that genotoxicity of ZnO and Ag NMs leading to DNA damage and alternatively to apoptosis in THP-1 macrophages is probably caused by the extensive intracellular dissolution of these NPs, as confirmed by TEM imaging.
Keywords:
nanomaterials; toxicity; THP-1 macrophages; gene expression profiling
Available at various institutes of the ASCR
WHOLE-GENOME EXPRESSION ANALYSIS IN THP-1 MACROPHAGE-LIKE CELLS EXPOSED TO DIVERSE NANOMATERIALS
From the perspective of the immune system, nanomaterials (NMs) represent invading agents. Macrophages are immune cells residing in all organs and tissues as the first line of defense. Interactions of ...
SIZE AS AN IMPORTANT FACTOR IN NANO-TiO2 TOXICITY IN MACROPHAGE-LIKE CELLS
Líbalová, Helena; Sikorová, Jitka; Brzicová, Táňa; Milcová, Alena; Vrbová, Kristýna; Pikal, P.; Topinka, Jan; Rössner ml., Pavel
2018 - English
A set of NPs consists of 5 variants of anatase and 5 variants of rutile nanoparticles differing in their diameter (from 3 to 165 nm). TiO2 samples were characterized in the powder form and dispersed in water and cell culture media. Three cytotoxicity assays were used: MTS, WST-1, and LDH. For all nanomaterials, three independent repetitions were carried out. \n\nOverall, cytotoxicity of all NPs was low even at the highest concentration of 256 mu g/ml. The viability of cells did not decrease below 60% for WST-1 and MTS assays and 80% for the LDH assay. Besides concentration, crystalline size was identified as the most important cytotoxic factor. Clear nonlinear relationship between crystalline size and cytotoxicity was detected, higher toxicity induced NPs within the size range 20-60 nm. Increased cytotoxicity in given diameter size range would give an answer to inconsistent findings at size and cytotoxicity relationship.
Keywords:
TiO2; nanoparticles; macrophages; cytotoxicity
Available at various institutes of the ASCR
SIZE AS AN IMPORTANT FACTOR IN NANO-TiO2 TOXICITY IN MACROPHAGE-LIKE CELLS
A set of NPs consists of 5 variants of anatase and 5 variants of rutile nanoparticles differing in their diameter (from 3 to 165 nm). TiO2 samples were characterized in the powder form and dispersed ...
Nanofiber scaffolds for local delivery of stem cells and immunosuppressive drugs for therapeutic purposes
Zajícová, Alena; Kössl, Jan; Heřmánková, Barbora; Boháčová, Pavla; Holáň, Vladimír
2018 - English
Cell-based therapy of local tissue injuries or damages requires application of stem cells and inhibition of harmful inflammatory reaction which could impede the healing process. To increase the effectiveness of this therapy, a local administration of drugs can avoid their side effects associated with a systemic treatment. A local therapy requires suitable carriers, which can transfer the cells and drugs to the site of injury. As a promising carriers turned out nanofiber scaffolds prepared by electrospinning technology from various types of polymers. The main advantage of this technology is a possibility to define properties of nanofiber scaffolds, optimal for the growth and transfer of stem cells, and which could incorporate various types of immunosuppressive drugs. Here we describe the formation and use of nanofiber scaffolds prepared by needleless electrospinning technology from poly (L-lactic acid) (PLA) which are loaded with immunosuppressive drug Cyclosporine A (CsA). We show that CsA-loaded nanofibers effectively and selectively inhibit proliferation of activated T cells and suppress the production of T cell cytokines in vitro. Simultaneously, these nanofiber scaffolds enable growth of mesenchymal stem cells (MSCs) and thus can serve as stem cell carriers. Moreover, using an experimental mouse model of skin transplantation, we showed that covering skin allografts with MSC-seeded and CsA-loaded nanofibers significantly inhibited the local production of pro-inflammatory cytokines IL-2, IL-17 and IFN-gamma, and supported healing. Thus, nanofiber scaffolds seeded with stem cells and loaded with CsA can serve as carriers of cells and drugs for a local cell therapy and for simultaneous effective immunosuppression.
Keywords:
nanofiber scaffold; immunosuppressive drug CsA; cytokines; stem cells
Available at various institutes of the ASCR
Nanofiber scaffolds for local delivery of stem cells and immunosuppressive drugs for therapeutic purposes
Cell-based therapy of local tissue injuries or damages requires application of stem cells and inhibition of harmful inflammatory reaction which could impede the healing process. To increase the ...
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