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Regeneration and structural changes of the nerve tissue after the extracellular matrix modification
Vallová, Jana ; Machová Urdzíková, Lucia (advisor) ; Hampl, Aleš (referee) ; Hubálek Kalbáčová, Marie (referee)
Regeneration and structural changes of the nerve tissue after the extracellular matrix modification. Abstract ECM modification may contribute to changes in nerve tissue plasticity. Therefore, 4- methylubulliferone is used in our study, which disrupted the structure of the perineuronal networks surrounding some types of neurons responsible for the formation of memory traces. Mice were fed a 4-MU diet (6.7 mg / g / day) for 6 months, which improved their memory skills in a spontaneous recognition test without a significant adverse effect on the kidneys, liver, or joints.In the next part of the study, an extracellular matrix (UC-ECM) was derived from fetal human umbilical cord tissue also generated as biomimetic hydrogel. Due to the generated UC-ECM's low stability and rapid degradation, the structure was stabilized by covalent genipin bonding. Stabilization with 1 mM genipine increased the biological stability of the material. UC-ECM as well as ECM/G didn't show toxicity in vitro in mesenchymal stem cell proliferation; axonal budding or neural stem cell growth and differentiation were not adversely affected. The biocompatibility of both materials was verified in vivo by applying the material to an intracortical photothrombotic rat lesion, where gelation and infiltration of the lesion and hydrogel by host...
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Studies of properties of viral capsid proteins and development of recombinant vaccines and diagnostic components based on artificial viral structures
Fraiberk, Martin ; Forstová, Jitka (advisor) ; Hubálek Kalbáčová, Marie (referee) ; Hejnar, Jiří (referee)
The aim of this study was to develop a system for easy production of different veterinary chimeric vaccines based on stable mouse polyomavirus (MPyV) structures. The system is designed for antigens that are problematic in production or stability. First, universal vectors for baculovirus-directed production of chimeric MPyV VLPs or pentamers based on the major capsid protein VP1 were designed to be exploited as vaccines against other pathogens. The different strategies used in this study are based on: A) exposure of selected immunogenic epitopes on the surface of MPyV VLPs by inserting them into a surface loop of the VP1 protein, B) insertion of foreign protein molecules inside the VLPs, or C) fusion of a foreign protein or its part with the C-terminus of VP1 protein, thus forming giant pentamers of a chimeric protein. Candidate vaccine antigens against porcine circovirus 2 (PCV2), the causative agent of porcine circovirus 2 systemic diseases (PCV2-SD) which causes significant economic losses in swine breeding, were prepared using the constructed vectors. All candidate vaccines induced the production of antibodies against the capsid protein of PCV2 after immunization of mice. The candidate vaccine Var C based on fusion of MPyV and PCV2 capsid proteins, is able to induce production of antibodies with...
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The effect of carbon nanostructures on human cell behavior and the role of fetal bovine serum in cell adhesion
Verdánová, Martina ; Hubálek Kalbáčová, Marie (advisor) ; Brábek, Jan (referee) ; Smetana, Karel (referee)
Graphene (G) and nanocrystalline diamond (NCD) are carbon allotropes and promising nanomaterials with an excellent combination of their properties, such as high mechanical strength, electrical and thermal conductivity, possibility of functionalization and very high surface area to volume ratio. For these reasons, G and NCD are employed next to electronics in biomedical applications, including implant coating, drug and gene delivery and biosensing. For a fundamental characterization of cell behavior on G and NCD, we studied osteoblast adhesion and proliferation on differently treated G and NCD. Generally, both G and NCD exhibited better properties for osteoblast cultivation than control tissue culture polystyrene. Better cell adhesion but lower cell proliferation were observed on NCD compared to G. The most surprising finding was that hydrophobic G with nanowrinkled topography enhanced cell proliferation extensively, in comparison to hydrophilic and flat G and both NCDs (hydrophobic and hydrophilic) with slightly higher roughness. Promoted cell proliferation enables faster cell colonization of G and NCD substrates, meaning faster new tissue formation which is beneficial in biomedical applications. Furthermore, it was shown that osteoblast adhesion was promoted in the initial absence of fetal bovine...
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The Cell-Nanomaterial Interactions and Their Application in Biomedicine
Vrabcová, Lucie ; Hubálek Kalbáčová, Marie (advisor) ; Pešta, Martin (referee) ; Benson, Veronika (referee)
Nanomaterials entered the biomedicine already at the beginning of the millennium and they still bring new and unique advances and possibilities for treatment, diagnosis and regeneration, thus they continue to facilitate the development of personalized medicine. Interactions at nanoscale allow until then unconceivable opportunities to influence processes on molecular level. The completely new perspectives of nanomaterial applications jointly raise attention concerning health and environmental safety issues. Although a lot of novel biomedical applications of nanomaterials have emerged, the vast majority is still found to be at the stage of a concept. The consistent basic in vitro research of elemental interactions of nanomaterial with biological environment should represent an essential part of its development. The concern of this thesis was to describe the cytocompatibility and interactions of two types of nanomaterials with different human cells. First, ultra-fine grain titanium was tested for prospective use in implant development. We confirmed its positive effect mainly on the growth of osteoblasts and recommended the further pre-clinical trials of this material in a form of a bone or dental implant. Second, several types of ultra-small (˂ 5 nm) nanoparticles of different origin (silicon, gold...
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Adhesion, growth and differentiation of osteoblasts and mesenchymal stromal cells on biocompatible nanomaterial surfaces
Brož, Antonín ; Hubálek Kalbáčová, Marie (advisor) ; Černý, Jan (referee) ; Kylián, Ondřej (referee)
The thesis is based on articles describing the fundamental research of carbon based nanomaterials for their possible utilization in biomedicine. The aim of this thesis was to describe the way how human osteoblasts (SAOS-2 cell line) and primary human mesenchymal stem cells (hMSC) adhere, grow and behave on surfaces made of several carbon allotropes - nanocrystalline diamond (NCD), single walled carbon nanotubes (SWCNTs) films and graphene. The utilization of carbon as the basic material promised good biocompatibility and possibility of useful surface modifications. The NCD had modified surface nanotopography (nanoroughness and nanostructuring prepared by dry ion etching). All the materials had modified surface atomic termination with oxygen and hydrogen which changes the surface electrical conductivity, surface charge and wettability. It was hypothesized that the surface termination can also influence the cell adhesion and growth. It turned out that all the studied materials were suitable as substrates for cultivation of mentioned cell types. Various nanoroughnesses of NCD surface had different effect on the cell adhesion and cell metabolic activity. Nanostructuring of the NCD influenced the formation of focal adhesions. The surface terminations of NCD and the other studied nanomaterials in...
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L1CAM and its role in cellular senescence
Mrázková, Blanka ; Hodný, Zdeněk (advisor) ; Hubálek Kalbáčová, Marie (referee) ; Rohlena, Jakub (referee)
Cellular senescence, originally defined as irreversible cell cycle arrest, was shown to act in organism as a double-edged sword. On the one hand, cellular senescence is considered an anti-cancer barrier and it beneficially contributes to processes such as wound healing and tissue regeneration. On the other hand, its longer persistence in the organism, usually when not eliminated by the immune system in elder age, cellular senescence contributes to age-related diseases and ageing itself. Moreover, senescent cells emerge as a result of radio- and chemo- therapy and can lead to detrimental effects when not eliminated. There is also accumulated evidence that senescent cells can overcome the proliferation barrier and become malignant (often after a cancer therapy) rendering senescence original definition invalid. In effort to improve the quality of health and life and to minimize the cancer risk after therapies, senescent cells have become one of the most interesting subjects for a targeted therapy. There is a promising potential in developing effective tools, namely drugs specifically killing senescent cells or reducing their detrimental effect (senolytics) that focus on senescent cells elimination in order to rejuvenate the organism, to extend a life span, and to prevent ageing-associated diseases and...
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The biocompatibility and potential cytotoxicity of materials for joint replacement manufacturing and coating
Kopová, Ivana ; Bačáková, Lucie (advisor) ; Hubálek Kalbáčová, Marie (referee) ; Jendelová, Pavla (referee)
Currently used prostheses for total joint replacement still have numerous disadvantages: extreme stiffness or elastic modulus of the bulk metallic material; insufficient integration of the implant into the host bone; and a high wear and corrosion rate, which causes an accumulation of mostly metallic or polymeric wear debris. Because of these reasons, many patients experience increasing local pain, swelling, allergic reactions, and inflammation resulting in bone loss and the aseptic loosening of the implant leading to the need for painful and expensive revision surgery. To address the mechanical issues of commonly used orthopaedic alloys, this thesis presents the development of the new β-type titanium alloy Ti-35Nb-7Zr-6Ta-2Fe-0.5Si with a relatively low elastic modulus (up to 85 GPa), increased tensile strength (880 MPa), and enhanced biocompatibility and osteoconductivity. Considering the generally low osteoinductivity of metallic implants, various surface modifications and coatings have been developed to improve the cell-material interaction, e.g. carbon-based coatings. Among these coatings, C60 fullerene layers have emerged as a great candidate for coating orthopaedic implants due to their therapeutic potential in arthritis. The potential cytotoxicity and DNA damage response of fullerenes have...
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Application on nanoparticles in bio-medicine.
Bělinová, Tereza ; Hubálek Kalbáčová, Marie (advisor) ; Beranová, Jana (referee)
During last few years nanoparticles of different origin have been used in biomedicine. Their interactions with cells are however a big point of concern and so further research is needed to be done regarding their properties, internalization into cells and their fate within the cell. All of this is needed for correct selection of proper nanoparticles for bioimaging and also for future use in human medicine. One of the main parts of research is deep understanding of interactions of nanodiamonds and silicon based nanoparticles with cells, because of their highly unique properties (autofluorescence), biocompatibility and in case of silicon nanoparticles also their degradability.
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Interactions of cells with nanoparticles for bio-medical applications
Bělinová, Tereza ; Hubálek Kalbáčová, Marie (advisor) ; Benson, Veronika (referee) ; Topinka, Jan (referee)
In the past decades, nanoparticles have been viewed as a potentially powerful platform for various applications in biomedical sciences. The possible application of nanoparticles varies from drug delivery agents to novel imaging platforms and surely, some application potential still remains hidden. Thus, it is necessary to broadly study their in vitro behavior in order to assess the precise theranostic potential as well as to distinguish possible threats to human health. Even though nanoparticles are getting more and more attention in current research, still only a limited amount of information is available, especially regarding interactions of ultra-small (< 5 nm) nanoparticles with biological environment and cells. The aim of the work presented herein is to provide the reader with information concerning interactions of various ultra-small nanoparticles (silicon-based, gold, nanodiamonds) with biological environment and human cells. Dose- and time-dependent influence of the various nanoparticles on behavior of different human cells (osteoblasts, monocytes, keratinocytes, mesenchymal stem cells) was established under different conditions, stressing out the importance of protein corona (a layer of proteins originating from cultivation medium attached to nanoparticles). Biocompatibility of two...
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The role of extracellular matrix in mesenchymal stem cell biology
Šimková, Zuzana ; Chudíčková, Milada (advisor) ; Hubálek Kalbáčová, Marie (referee)
The aim of this bachelor thesis is to describe the mutual relations of mesenchymal stem cells and extracellular matrix, their communication and to point out their significance in living body. First part is focused on description of extracellular matrix, its components and characteristics. The interaction between extracellular matrix and surrounding cells is using both receptors and enzymes. Second part addresses the description of mesenchymal stem cells which are crucial producers of extracellular matrix. The matrix is also decisive part of stem cells niche which envelopes them, protects them and decides their future. The last section demonstrates possible utilizations of differentiation potential and paracrine secretion of MSC in therapy and identifies some of the risks bound to it.
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