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Targeting of viral nanoparticles to cancer specific receptors
Žáčková Suchanová, Jiřina ; Španielová, Hana (advisor) ; Němečková, Šárka (referee) ; Ulbrich, Pavel (referee)
The aim of this thesis is to reveal the potential of mouse polyomavirus (MPyV) based virus-like particles (VLPs) as possible nanocarriers for directed delivery of therapeutic or diagnostic compounds to specific cells or tissues. We have chosen mouse polyomavirus VLPs because they do not contain viral DNA and are considered safe for utilization in bio-applications. In our research, we used a chemical approach for retargeting of MPyV based VLPs from their natural receptor to cancer cells. The chemical modification of the capsid surface exposed lysines by an aldehyde-containing reagent enabled conjugation of VLPs to selected molecules: transferrin and inhibitor of glutamate carboxypeptidase II (GCPII). Transferrin, as a transporter of iron to metabolically active cells, targeted VLPs to numerous types of cancer cells overexpressing the transferrin receptor. On the other hand, GCPII serves as a transmembrane marker specific for prostate cancer cells and conjugation of its inhibitor to VLPs resulted in successful recognition of these cells. Electron microscopy was used for visualization of modified VLPs and flow cytometry together with confocal microscopy for investigation of cell specific interactions and VLP uptake. Furthermore, we explored the influence of serum proteins on VLPs. The abundance of...
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Coat protein-RNA interaction in vivo and the biotechnological use of VLPs
Kratochvílová, Kateřina ; Moravec, Tomáš (advisor) ; Hála, Michal (referee)
The Tobacco mosaic virus (TMV) is a simple and frequently used model virus which has been studied already more over than 130 years. Due to the intensive study of this virus the details of its infectious cycle, genomic information and also the structure of the created viral particle as well as the mechanism of its creation are known today. The process of encapsidation (viral particle formation) is sufficiently described in the in vitro conditions. In the in vitro conditions the origin of assembly (OAS) was also described. The OAS was identified in the coding sequence of the gene for the movement protein (MP). The importance of replication centers (replication factories) has also been supposed. The aim of the diploma thesis was to study the specificity of the interaction of RNA and coat protein in the process of the particle assembly taking place directly inside the plants. The experiments were performed to verify the necessity of presence of OAS sequence in process of initiation of viral encapsidation. The effect of the cell compartmentation on this process has also been studied. Based on several viral systems (the Tobacco mosaic virus, the Potato virus X, the Bean yellow dwarf virus and Cowpea mosaic virus) gene constructs were created. These constructs enables to study this idea at the molecular...
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Design and characterization of advanced polymer-coated upconversion nanoparticles
Kostiv, Uliana ; Horák, Daniel (advisor) ; Kaman, Ondřej (referee) ; Brožek, Jiří (referee)
Lanthanide-based upconversion nanoparticles are able to absorb low-energy near-infrared (NIR) photons and emit visible or ultraviolet light. This unique optical property enables luminescent detection without autofluorescence from biological samples that is crucial for bioimaging and diagnostics. Moreover, NIR irradiation allows to deliver light deep into the tissue, which can be used for drug or gene delivery and NIR-induced photodynamic therapy (PDT) of tumors. Despite huge effort during the last 10 years devoted to the upconversion nanoparticle synthesis and their surface engineering, preparation of the particles suitable for biomedical applications still remains a big challenge. The current work is focused on synthesis of the upconversion nanoparticles with controllable morphology, size, distribution, crystallinity, and high upconversion efficiency, as well as on desirable surface modification by biocompatible inorganic or organic polymers. Neat and functionalized homogeneous silica shell was introduced on the NaYF4:Yb3+ /Er3+ and NaGdF4:Yb3+ /Er3+ nanoparticles to enhance their biocompatibility and colloidal stability in water. To render the nanoparticles with targeting moieties, their surface was decorated with cell-adhesive RGD or cell-penetrating TAT peptides. The RGD- and TAT- conjugated...
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Preparation of uniform superparamagnetic particles with polymer coating for biomedical applications
Patsula, Vitalii ; Horák, Daniel (advisor) ; Sysel, Petr (referee) ; Matějíček, Pavel (referee)
Aim of this thesis was to design and prepare polymer-coated monodisperse Fe3O4 nanoparticles as a safe and non-toxic contrast agent for magnetic resonance imaging (MRI) and heat mediator for hyperthermia. Uniform superparamagnetic Fe3O4 nanoparticles were synthesized by thermal decomposition of Fe(III) oleate, mandelate, or glucuronate in high- boiling solvents at temperature >285 řC. Size of the particles was controlled in the range of 8- 27 nm by changing reaction parameters, i.e., temperature, type of iron precursor, and concentration of stabilizer (oleic acid and/or oleylamine), while preserving uniformity of the nanoparticles. Because particles contained hydrophobic stabilizer on the surface, they were dispersible only in organic solvents. To ensure water dispersibility, oleic acid on the particle surface was replaced by hydrophilic and biocompatible methoxy-poly(ethylene glycol) (PEG) and poly(3-O-methacryloyl-α-D-glucopyranose) by ligand exchange. Polymers were previously terminated with anchoring-end groups (hydroxamic or phosphonic) to provide firm bonding to iron atoms on the particle surface. Fe3O4 nanoparticles were also hydrophilized by encapsulation into a silica shell by reverse microemulsion method. Tetramethyl orthosilicate was used to prepare Fe3O4@SiO2 nanoparticles, which were...
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Influence of size and geometry of nanoparticles on cellular internalization pathways
Číhařová, Barbora ; Španielová, Hana (advisor) ; Lišková, Petra (referee)
Nanoparticles can be used in biomedical disciplines as carriers for transport of diagnostic as well as therapeutic substances into cells. Variety of different shapes, sizes and different compositions are used experimentally. Despite the discoveries already made in this area, the exact nature of the interaction between a nanoparticle and a cell has not been fully understood yet. The objective of this thesis is to provide the knowledge about possibilities of utilisation and aspects influencing the interaction between the cell membrane and several types of nanoparticles: liposomes, gold nanoparticles and virus-like nanoparticles. The comparison shows that generalisation of the mechanism of nanoparticle entry into the cell is problematic, although it seems that the spherical nanoparticles with the diameter of 50 nm provide the most efficient entry.
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Plasma polymers in the nanostructured and nanocomposite coatings
Shelemin, Artem ; Biederman, Hynek (advisor) ; Čech, Vladimír (referee) ; Vyskočil, Jiří (referee)
Title: Plasma polymers in the nanostructured and nanocomposite coatings Author: Artem Shelemin Department / Institute: Department of the Macromolecular Physics Supervisor of the doctoral thesis: Prof. RNDr. Hynek Biederman, DrSc. Abstract: The thesis represents the main results of my research work aimed to study nanostructured and nanocomposite films of plasma polymer. A few alternative experimental approaches were developed and investigated which ranged from low pressure (gas aggregation cluster sources and glancing angle deposition) to atmospheric pressure (dielectric barrier discharge and plasma jet) plasma processing. The metal/metal oxide Ti/TiOx, AlOx and plasma polymer SiOx(CH), Nylon 6,6 nanoparticles were prepared. The analysis of morphology of deposited plasma polymer coatings was performed by AFM and SEM. The chemical composition of films was studied by XPS and FTIR. Keywords: plasma polymer, nanoparticle, thin film, nanostructures
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Preparation of graphene quantum dots and study of their properties
Zdražil, Lukáš ; Mikula, Milan (referee) ; Zmeškal, Oldřich (advisor)
Current methods for preparing graphene quantum dots (GQDs) rely on oxidation and reducing agents or require energy-intensive and technologically demanding preconditions. Applying microwave expansion and liquid phase exfoliation (LPE) in a sample of graphite powder enabled us to prepare GQDs that exhibit strong luminescence in the blue region of the visible spectrum. The proposed technique for synthesizing GQDs is energetically undemanding and does not necessitate additional chemical components.
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The issue of the use of carbon nanotubes in cement composites
Pacltová, Klára ; Hela, Rudolf (referee) ; Bodnárová, Lenka (advisor)
The bachelor thesis deals with the analysis of the efficiency of carbon nanoparticles in cement composites. The theoretical part of the bachelor thesis is focused on the research of information about carbon nanotubes and their influence on cement composites. The practical part follows the research of the theoretical part, where is verified the effectiveness of carbon nanotubes in cement composites at different dosages.
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Preparation and characterization of diamond-based nanocarriers for transfection of siRNA
Majer, Jan ; Cígler, Petr (advisor) ; Fišer, Radovan (referee)
Although nanodiamonds were discovered and produced tens of years ago, they have been utilized in medical and biological fields just recently, particularly in drug and gene delivery into a cell and in bioimaging methods. Nanodiamonds can be modified with specific positively charged moieties for complexation with negatively charged nucleic acids. These complexes afterwards overcome extracellular and intracellular barriers and transport the nucleic acid either into cytosol or into the nucleus. Owing to fluorescence centres nitrogen- vacancy, which can be formed in the nanodiamonds, nanodiamonds exhibit excelling optical properties, as they emit stable fluorescence without "photoblinking" or "photobleaching". This thesis reviews properties, synthesis and modifications of nanodiamonds and other selected nanoparticles and their in vitro applications. This thesis also compares their cytotoxicity and gene knockdown efficiency.
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