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The co-culture of keratinocytes and fibroblasts on a multi-layered polyester nanofibrous membrane enriched with platelet lysate
Blanquer, Andreu ; Filová, Elena ; Jenčová, V. ; Brož, Antonín ; Kuželová Košťáková, E. ; Lisnenko, M. ; Procházková, R. ; Bačáková, Lucie
The prevalence of chronic wounds is increasing due to the population ageing and specific illnesses like diabetes mellitus and vascular diseases. Nanofibrous membranes fabricated using synthetic polymers are promising materials to enhance skin wound healing. PCL and PVA membranes are being studied to be used as scaffolds for skin tissue engineering and hydrogels for controlled drug delivery, respectively. The present study considers the development of a multi-layered membrane made of PCL and PVA loaded with platelet lysate (PL). PCL nanofibers allowed cell adhesion and growth, whereas PVA acted as a hydrogel that releases the bioactive compounds of platelet lysate. The cytocompatibility of the membranes containing PL and without it was demonstrated on two cell types involved in wound healing, i.e. keratinocytes and fibroblasts. Both cell types were able to adhere and proliferate on the membranes. In addition, the membrane containing PL enhanced the proliferation of fibroblasts. A co-culture study was also performed by seeding each cell type on one side of the membrane. The cells were co-cultured for 7 days and the results showed that PL increased the proliferation of cells achieving a monolayer of keratinocytes or fibroblasts on each side of the membrane. Thus, the beneficial effect of PCL-PVA+PL membranes on monocultures and co-cultures of skin cells was demonstrated, and these membranes can be considered potential scaffolds for treatment of chronic wounds.
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Degradation of polycaprolactone electrospun materials - methods of analysis
Havlíčková, K. ; Řezanka, M. ; Kobera, Libor ; Kuželová Košťáková, E. ; Lukáš, D. ; Jenčová, V.
Biodegradable electrospun materials are widely used for medical application. Polycaprolatone is polymer suitable for electrospinning technology and is very often used to create nanofibrous scaffolds for tissue engineering. The time to disintegration or biodegradation of such materials is very important here. However, testing is not entirely easy. It is not possible to create exactly body-like conditions in vitro. Moreover, it is not easy to find suitable analytical methods that would show exactly what happens in the nanofibrous polycaprolactone electrospun samples at certain stages of degradation, ie how the internal structure of decaying nanofibers changes. This paper describes the traditional use of methods for testing polycaprolatone nanofibers by enzymatically catalysed degradation. Morphological changes are studied using scanning electron microscope images. However, it also offers a non-traditional analysis of polycaprolactone electrospun materials using the ssNMR method.
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A COMPARISON OF THE CENTRIFUGAL FORCE SPINNING AND ELECTROSPINNING OF COLLAGEN UNDER DIFFERENT CONDITIONS
Hlavatá, J. ; Suchý, Tomáš ; Šupová, Monika ; Pokorný, M. ; Košťáková, E.
Collagen makes up one of the most important parts of the extracellular matrix and connective tissue. Collagen type I accounts for up to 90% of the collagen found in the body and it can be isolated and purified in large quantities and processed into a variety of forms. With respect to the process of the treatment of collagen into the nano-fibrous and submicron-fibrous forms, severe conditions leading to the denaturation of the collagen and thus the gelatin content should be avoided. Centrifugal force spinning and electrospinning processes appear to present promising methods for the spinning of collagen solutions. This paper deals with a comparison of the various methods applied in the processing of collagen, i.e. principally needle and needleless centrifugal spinning technology and electrospinning technology. In addition, the effect of process parameters and collagen solution conditions on the final form of the material was analyzed. Spun layers were prepared based on collagen type I isolated from calf skin. Nano-structured layers were prepared employing the spinning of 4-16 wt% collagen solutions in phosphate buffer saline and ethanol. The layers thus prepared were characterized by means of scanning electron microscopy and Fourier transform infrared spectroscopy.
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Budgeting of Taxes
Houdková, Kamila ; Sedmihradská, Lucie (advisor) ; Košťáková, Eliška (referee)
This diploma thesis examines the impact of the amendment to the Act on Budgeting of Taxes, effective from 1 January 2013, on the financing of municipalities setting up a school in their territory. The main aim of the diploma thesis is to find out what influence the amendment to the School Act and the Act on the Budgeting of Taxes has on the financing of municipalities establishing school and municipalities that do not establish school in their territory. Furthermore, whether the municipality, which establishes a school in its territory, is able to cover the costs incurred by education when fees are abolished, and the pupils from the municipalities in whose territory the school is not established are also able to attend. The diploma thesis deals with the issue of financing the municipality, which establishes a school and a municipality that does not establish a school in its territory. Then the issue is related to the whole Pardubice region, where I find out whether the municipalities setting up the school in their territory will be able to cover the costs related to the operation of the school after the amendment of the Act on Budgeting of Taxes.
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In vitro testing of carrier system based on nanofibres for vitiligo treatment
Kodedová, Barbora ; Amler, Evžen (advisor) ; Kuželová Košťáková, Eva (referee)
Vitiligo is a skin disease with 2 % prevalence in a worldwide population. It is characterised by loss or decrease in activity of epidermal melanocytes, which lead to skin and hair depigmentation. It has negative impact on psyche, social relationships of patients and reduces the protection of the organism against UV radiation. One of the treatment methods is autologous transplantation of melanocytes or suspension of melanocytes with keratinocytes. Use of the biocompatible membrane, which allows the cultivation of these cells with resulting transplantation on the depigmented lesion, could improve treatment and make it more efficient. The main goal of this work was to create the biocompatible membrane from nanofiber layers of polyvinylalcohol (PVA) which can stand as a carrier for cell transplants in vitiligo therapy. PVA scaffolds were prepared by electrostatic spinning and later on modified by cold methane plasma (CH4) for lowering their hydrofility. Samples of modified nanofiber carriers were analysed according to their physical and chemical characteristics (visualization fiber morphology by SEM, XPS and surface Zeta potential analysis and contact angle). Consequently, adhesion, proliferation and metabolic activity of cultivating mice cell lines of melanocytes (Melan-a) and keratinocytes (XB2) were examined...
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Tax Evasion and Transfer Pricing
Košťáková, Eliška ; Vítek, Leoš (advisor) ; Zídková, Hana (referee)
The aim of the diploma thesis is to answer the question of where is the line between tax evasion, tax avoidance and tax fraud. Furthermore to characterize methods of measurement tax evasion and introduced actions against tax evasion and tax avoidance at the national and international level. This aim is achieved through analytical and synthetic methods used from the first to third chapter of the diploma thesis. The main goal of diploma thesis is to use external sources of information to determining transfer pricing methods in small economic models. To achieve this goal is at first step explain the Arm's length principle, comparative analysis and subsequently introduced each transfer pricing method. The attention is also paid to assessment of each transfer pricing method with using analytical, synthetic and comparative methods.
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