Number of found documents: 140
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Optimalizace kultivačních podmínek lidských jaterních buněk HepG2 na 4 typech nano- a micro- vlákenných nosičů
Rössner ml., Pavel
2020 - Czech
Technologie popisuje optimalizaci počtu nasazovaných lidských hepatocytů HepG2, vhodných pro využívání v oblasti toxikologického ověřování nových léčiv či potravin, na 3D kultivační systém tvořený čtyřmi typy nano- a mikrovlákenných nosičů. Motivací pro hledání vhodných modelů testování potenciálně genotoxických účinků léčiv je nízká relevance tkáňových modelů užívaných v prvních fázích preklinického hodnocení nových látek a také redukce testování na zvířatech. The technology describes the optimization of the number of deployed human hepatocytes HepG2, suitable for use in the field of toxicological verification of new drugs or foods, on a 3D culture system consisting of four types of nano- and microfiber carriers. The motivation for finding suitable models for testing potentially genotoxic effects of drugs is the low relevance of tissue models used in the first stages of preclinical evaluation of new substances, as well as the reduction of animal testing. Keywords: nanofiber carriers; toxicology; HepG2 Available at various institutes of the ASCR
Optimalizace kultivačních podmínek lidských jaterních buněk HepG2 na 4 typech nano- a micro- vlákenných nosičů

Technologie popisuje optimalizaci počtu nasazovaných lidských hepatocytů HepG2, vhodných pro využívání v oblasti toxikologického ověřování nových léčiv či potravin, na 3D ...

Rössner ml., Pavel
Ústav experimentální medicíny, 2020

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 digital repository of the ASCR
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 ...

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
Ústav experimentální medicíny, 2020

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
2020 - 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 ...

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
Ústav experimentální medicíny, 2020

Modifikovaná metoda hodnocení mikrojader v lymfocytech s použitím hybridizace pancentrometrických fluorescenčně značených sond
Rössner ml., Pavel; Rössnerová, Andrea
2020 - Czech
Tato metodika je založena na současném fluorescenčním barvení jak celých chromozomů, tak jejich centromer pomocí pancentrometrických sond. Díky tomu je možné vyhodnotit frekvenci mikrojader s / bez centromer, tedy určit rozdíly mezi strukturálními a numerickými aberacemi. Cílem metodiky bylo vytvořit inovativní postup pro vyhodnocení genotoxických účinků nanočástic, chemických látek včetně léků či záření na DNA živých organismů. This methodology is based on the simultaneous fluorescent staining of both whole chromosomes and their centromere using pancentrometric probes. Thanks to this, it is possible to evaluate the frequencies of micronuclei with / without centromeres, ie to determine the differences between structural and numerical aberrations. The aim of the methodology was to create an innovative procedure for evaluating the genotoxic effects of nanoparticles, chemicals, including drugs or radiation on the DNA of living organisms. Keywords: genotoxicity; fluorescent staining; pancentrometric probes; nanoparticles Available at various institutes of the ASCR
Modifikovaná metoda hodnocení mikrojader v lymfocytech s použitím hybridizace pancentrometrických fluorescenčně značených sond

Tato metodika je založena na současném fluorescenčním barvení jak celých chromozomů, tak jejich centromer pomocí pancentrometrických sond. Díky tomu je možné vyhodnotit frekvenci mikrojader s / bez ...

Rössner ml., Pavel; Rössnerová, Andrea
Ústav experimentální medicíny, 2020

Metodika využití nanovlákenných nosičů a kmenových buněk pro léčbu závažných poškození oka
Zajícová, Alena; Javorková, Eliška; Holáň, Vladimír
2019 - Czech
Metodika popisuje nový léčebný postup pro léčbu závažných poškození očního povrchu ve veterinární medicíně. Postup je založen na kultivaci kmenových buněk a na jejich přenosu pomocí nanovlákenných nosičů na poškozený oční povrch. Metoda je využitelná v případech, kde již jiné dostupné formy léčby nejsou úspěšné. The method describes a new therapeutic approach for the treatment of severe ocular injuries in veterinary medicine. The protocol is based on a cultivation of stem cells and their transfer using nanofiber scaffolds onto damaged ocular surface. This method can be used in the cases when other available treatment options are not sufficient or cannot be used. Keywords: nanofiber carriers; stem cells; ocular surface injuries; therapeutic methods; veterinary medicine Available at various institutes of the ASCR
Metodika využití nanovlákenných nosičů a kmenových buněk pro léčbu závažných poškození oka

Metodika popisuje nový léčebný postup pro léčbu závažných poškození očního povrchu ve veterinární medicíně. Postup je založen na kultivaci kmenových buněk a na jejich přenosu pomocí nanovlákenných ...

Zajícová, Alena; Javorková, Eliška; Holáň, Vladimír
Ústav experimentální medicíny, 2019

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 ...

Jiráková, Klára; Moskvin, Maksym; Horák, Daniel; Jendelová, Pavla
Ústav experimentální medicíny, 2018

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 ...

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
Ústav experimentální medicíny, 2018

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 ...

Brzicová, Táňa; Líbalová, Helena; Vrbová, Kristýna; Sikorová, Jitka; Philimonenko, Vlada; Kléma, J.; Topinka, Jan; Rössner ml., Pavel
Ústav experimentální medicíny, 2018

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 ...

Líbalová, Helena; Sikorová, Jitka; Brzicová, Táňa; Milcová, Alena; Vrbová, Kristýna; Pikal, P.; Topinka, Jan; Rössner ml., Pavel
Ústav experimentální medicíny, 2018

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 ...

Zajícová, Alena; Kössl, Jan; Heřmánková, Barbora; Boháčová, Pavla; Holáň, Vladimír
Ústav experimentální medicíny, 2018

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