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Metodika využití nanovlákenných nosičů a kmenových buněk pro léčbu závažných poškození oka
Zajícová, Alena; Javorková, Eliška; Holáň, Vladimír
2019 - český
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. Klíčová slova: nanofiber carriers; stem cells; ocular surface injuries; therapeutic methods; veterinary medicine Plné texty jsou dostupné na jednotlivých ústavech Akademie věd ČR.
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 - anglický
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. Klíčová slova: neural stem cells; nanoparticles; oxidative stress; ascorbic acid Plné texty jsou dostupné na jednotlivých ústavech Akademie věd ČR.
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 - anglický
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. Klíčová slova: Zinc oxide nanoparticles; inhalation; gene expression; Immune response Plné texty jsou dostupné na jednotlivých ústavech Akademie věd ČR.
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 - anglický
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. Klíčová slova: nanomaterials; toxicity; THP-1 macrophages; gene expression profiling Plné texty jsou dostupné na jednotlivých ústavech Akademie věd ČR.
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 - anglický
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. Klíčová slova: TiO2; nanoparticles; macrophages; cytotoxicity Plné texty jsou dostupné na jednotlivých ústavech Akademie věd ČR.
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 - anglický
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. Klíčová slova: nanofiber scaffold; immunosuppressive drug CsA; cytokines; stem cells Plné texty jsou dostupné na jednotlivých ústavech Akademie věd ČR.
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 - anglický
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. Klíčová slova: BEAS-2B cells; genotoxicity; micronucleus assay; nanomaterials Plné texty jsou dostupné na jednotlivých ústavech Akademie věd ČR.
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 - anglický
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. Klíčová slova: Cornea; nanofiber scaffold; limbal stem cell deficiency; mesenchymal stem cells Plné texty jsou dostupné na jednotlivých ústavech Akademie věd ČR.
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

L01 DNA damage formation and DNA repair following an intervention of colorectal cell lines with ganoderma lucidum
Vodička, Pavel; Opattová, Alena; Čumová, Andrea; Slíva, D.
2016 - anglický
Colorectal cancer (CRC) is the third most common malignancy in the world and second most common cause of cancer related deaths in Europe. CRC is complex disease that develops as consequence of environmental and health risk factors with involvement of suboptimal DNA repair, resulting in an accumulation of DNA damage. Reactive oxygen species (ROS) are highly reactive molecules strictly controlled by cellular antioxidant system. Disturbance in the prooxidation–antioxidation homeostasis increases an extent of ROS and consequently an accumulation of DNA damage as well as apoptosis. \nMany natural compounds possess anti-cancer activities tentatively mediated by the generation of ROS. Cancer cells are more sensitive to oxidative DNA damage than non-malignant ones. Modulation of oxidative DNA damage and its repair by natural compounds may lead to selective cancer cell-death and further sensitization of cancer cells to the treatment. Ganoderma Lucidum (GLC) (Reishi, Ling-Zhi), a mushroom used in Chinese medicine for thousands of years, represents an example of a natural compound with empirically recorded anti-cancer as well as anti-proliferative effects. \nThe aim of our study is to define effect of Ganoderma lucidum (GLC) extract on DNA damage and DNA repair system in colorectal cell lines with different genetic backgrounds.\nOur results suggest that GLC extract decreases activity of the cellular antioxidant system which leads to oxidative DNA damage. GLC extract increases genotoxic burden in colorectal cancer cell lines, highlighted by the suppressed base excision repair capacity. These data indicate that specific oxidative DNA damage caused by natural compounds may become a potential tool for the improvement of specific anti-cancer treatment.\n Klíčová slova: colorectal cancer; natural compound; oxidative DNA damage; DNA repair; 5-fluorouracil; colorectal cell lines Plné texty jsou dostupné na jednotlivých ústavech Akademie věd ČR.
L01 DNA damage formation and DNA repair following an intervention of colorectal cell lines with ganoderma lucidum

Colorectal cancer (CRC) is the third most common malignancy in the world and second most common cause of cancer related deaths in Europe. CRC is complex disease that develops as consequence of ...

Vodička, Pavel; Opattová, Alena; Čumová, Andrea; Slíva, D.
Ústav experimentální medicíny, 2016

Natural compounds and their effect on 5-fluorouracil in colorectal cancer cell lines
Čumová, Andrea; Opattová, Alena; Vodenková, Soňa; Horák, Josef; Slíva, D.; Vodička, Pavel
2016 - anglický
Colorectal cancer (CRC) is the second most common type of cancer and the second most common cause of cancer related deaths in Europe. 5-Fluorouracil (5-FU) is widely used in treatment of various cancers including CRC, but apart from the cytotoxic effect on cancer cells may also cause adverse toxic side effects. 5-FU is an anti-metabolite with chemical structure similar to that of the pyrimidine molecules of DNA and RNA. However, response to chemotherapy is often limited by drug resistance. The p53 protein is one of the most widely studied tumour suppressors and mutations in TP53 gene are frequently detected in different types of tumours. \nGanoderma Lucidum (GLC) is a mushroom used in Traditional Eastern Medicine which exhibits anti-cancer and anti-proliferative effects in vitro\nThe aim of our study is to define the role of p53 in the interaction between 5-FU and GLC extract and their simultaneous effect on survival in CRC cell lines.\nOur results suggest that GLC extract significantly increases cytotoxicity and genotoxicity of 5-FU in CRC lines with different p53 status and may potentially modulate the response of p53 knock-out cells which are less sensitive to 5-FU treatment. Interaction of conventional chemotherapeutics with natural compounds introduces a novel aspect in cancer research and therapy.\n\n Klíčová slova: colorectal cancer; natural compound; p53; DNA repair; 5-fluorouracil; drug resistance Plné texty jsou dostupné na jednotlivých ústavech Akademie věd ČR.
Natural compounds and their effect on 5-fluorouracil in colorectal cancer cell lines

Colorectal cancer (CRC) is the second most common type of cancer and the second most common cause of cancer related deaths in Europe. 5-Fluorouracil (5-FU) is widely used in treatment of various ...

Čumová, Andrea; Opattová, Alena; Vodenková, Soňa; Horák, Josef; Slíva, D.; Vodička, Pavel
Ústav experimentální medicíny, 2016

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