Number of found documents: 138
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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

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

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

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

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

GENOTOXICITY OF NANOMATERIALS IN BEAS-2B CELLS ANALYZED BY THE IN VITRO MICRONUCLEUS ASSAY
Rössnerová, Andrea; Červená, Tereza; Brzicová, Táňa; Vrbová, Kristýna; Sikorová, Jitka; Topinka, Jan; Rössner ml., Pavel
2018 - English
The tremendous increase of the use of nanomaterials (NMs) has been witnessed during the last decade in many areas of human life including the chemical industry, cosmetics, biomedicine or food technology. The variety of NMs, their unique properties, almost ubiquitous presence and the size range of 1-100 nm raised the interest of toxicologists. The evaluation of the frequency of micronuclei (MN) as a result of the genotoxic events is a broadly utilized and well-established approach in in vitro studies for testing the risk of chemical exposure. Nevertheless, properties of the NMs give rise to the questions concerning the optimal methodological variants of the MN assay. \n\nIn our study, five types of well-characterized NMs (TiO2: NM-101 and NM-103; SiO2: NM-200; Ag: NM-300K and NM-302) of specific size, shape, or e.g. dimensions of aggregates were involved in the genotoxicity testing using four variants of protocols differing in the time of NM exposure, application of cytochalasin-B combined with simultaneous and delayed co-treatment with nanoparticles (NPs). Bronchial epithelial cells (BEAS-2B) were used in this study to fulfil these tasks. Presence of NPs was controlled by transmission electron microscopy (TEM). \n\nObtained results showed the different genotoxic potential of the various TiO2 and Ag NMs (NM-101< NM-103 and NM-300K> NM-302, respectively). Comparison of all testing strategies revealed, that the level of DNA damage can differ based on the time of exposure and the methodological approach. In general, using cytochalasin-B led most frequently to the increase of the genotoxic potential of the tested NMs. Keywords: BEAS-2B cells; genotoxicity; micronucleus assay; nanomaterials Available at various institutes of the ASCR
GENOTOXICITY OF NANOMATERIALS IN BEAS-2B CELLS ANALYZED BY THE IN VITRO MICRONUCLEUS ASSAY

The tremendous increase of the use of nanomaterials (NMs) has been witnessed during the last decade in many areas of human life including the chemical industry, cosmetics, biomedicine or food ...

Rössnerová, Andrea; Červená, Tereza; Brzicová, Táňa; Vrbová, Kristýna; Sikorová, Jitka; Topinka, Jan; Rössner ml., Pavel
Ústav experimentální medicíny, 2018

Use of the nanofiber scaffold for transfer of stem cells onto the injured ocular surface in mouse experimental model
Kössl, Jan; Zajícová, Alena; Heřmánková, Barbora; Javorková, Eliška; Boháčová, Pavla; Holáň, Vladimír
2018 - English
Corneal damage is one of the most common causes of impaired vision or even blindness. When the injury is more extensive and the limbal region is involved, the natural regeneration of the cornea is not sufficient. Such damage can lead to the limbal stem cell deficiency (LSCD). The only option for LSCD treatment is transplantation of the limbal tissue or a transfer of limbal stem cells (LSCs) cultured from the healthy eye. The allogenic transplantation of the limbus or cultivated LSCs with a systemic administration of immunosuppressive drugs is needed in the case of bilateral LSCD. Nevertheless, the cell therapy is very promising approach for LSCD treatment. Transplantation of mesenchymal stem cells (MSCs) seeded on an appropriate scaffold turned out to be a suitable therapy of the LSCD. In our experimental model of LSCD we use nanofiber scaffold for MSC and LSC cultivation and for transplantation of these cells onto the chemically injured mouse eye. MSCs have immunosuppressive and immunomodulatory properties. We showed that MSCs have the ability to inhibit production of molecules associated with the inflammation and support epithelial regeneration in the damaged cornea. These inhibitory properties were confirmed in both in vitro and in vivo mouse model. Results thus showed beneficial effects of stem cell transplantation for murine corneal healing and for suppression of a local immune reaction which can impede the healing process. Such similarity of in vivo and in vitro results allows us further experiments to clarify mechanisms of MSC regenerative and healing properties after the transplantation onto the injured cornea. Keywords: Cornea; nanofiber scaffold; limbal stem cell deficiency; mesenchymal stem cells Available at various institutes of the ASCR
Use of the nanofiber scaffold for transfer of stem cells onto the injured ocular surface in mouse experimental model

Corneal damage is one of the most common causes of impaired vision or even blindness. When the injury is more extensive and the limbal region is involved, the natural regeneration of the cornea is not ...

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

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