|
|
Development of Material Based on Hyaluronic Acid s Hydrogels for Myocardial Regeneration
Kovářová, Lenka ; Kubala,, Lukáš (referee) ; Prokš,, Vladimír (referee) ; Pekař, Miloslav (advisor)
The thesis is focused on material development based on hyaluronic acid usable in regenerative medicine, especially for heart tissue regeneration after myocardial infarction. The object of the study is the oxidized form of hyaluronic acid (HA-Ox) and hydroxyphenyl derivative of HA (HA-TA). HA-Ox can be crosslinked with a bifunctional alkoxyamine POA and HA-TA undergoes an enzymatic reaction in the presence of hydrogen peroxide catalysed by horseradish peroxidase leading to gel formation. To describe the materials, chemical and physical properties, gelation kinetics and conditions of crosslinking reactions were studied. Hydrogels were characterized by mechanical and viscoelastic properties, degradability or stability in simulated body fluids. These hydrogels serve as scaffolds for the selected cell type. To promote cell adhesion and viability, an RGD sequence has been bonded to the structure of HA-TA. This resulting material is also compatible with selected applicators. Its viscosity and extrusion force are low enough to allow application with a catheter with a very small internal diameter. The applicability of the material through the supply tube to the hydrogel reservoir of the second SPREADS device showed good homogeneity, cell distribution and viability. Finally, the material was applied in vivo using these devices during a preclinical study.
|
|
|
Simulation of adhesion and transmigration imune-cells through capillary wall
Morgaenko, Katsiarina ; Čmiel, Vratislav (referee) ; Chmelíková, Larisa (advisor)
Teoretická část této práce obsahuje biofyzikální popis endoteliální vrstvy a transmigrace buněk přes tuto vrstvu. Dále popisuje charakteristiky všech důležitých komponent pro vytvoření modelu cévy in vitro (endoteliální buňky, kmenové buňky, leukocyty, Calcein AM, PKH26). Praktická část práce je věnována sestavení in-vitro modelů dvojrozměrných a trojrozměrných endoteliálních vrstev, jejich zobrazení pomoci mikroskopu a testování interakce leukocytů a kmenových buněk s těmito endoteliálními vrstvami. V závěru je navrhnut nejlepší tvar a velikost mikrofluidních systémů simulujících kapiláry.
|
|
|
3D bioprinting of stem cells and analysis of microscopic images
Kandra, Mário ; Svoboda, Ondřej (referee) ; Kolářová, Jana (advisor)
In this diploma thesis we are discussing about using 3D bioprinting in tissue engineering. We are discribing using biomaterials for construction scaffolder and aplication stem cells in 3D bioprinting. Last section of theoretical part deals with very often used techniques of 3D bioprinting and we are focused on extrusion technique. In the practical part we propose a method for print vasculars structures. We realized prototype of print head, her design and 3D printing of individual parts. To mechanical part we create a control system for printing control. At the end we visualize the organization of the cells using program modules.
|
|
|
Proteomic analysis of posttranslation modifications in breast cancer cell line profiles
Predná, Nikola ; Laštovičková,, Markéta (referee) ; Strouhalová,, Dana (advisor)
Estrogenové a progesteronové receptory, stejně jako HER2 protein, jsou v současnosti klinicky nejužitečnějšími metabolickými markery u karcinomu prsu. Tyto markery umožňují určit typ nádoru a nejlepší možnosti jeho léčby. Jeden z nejagresivnějších typů tohoto onemocnění, triple-negative breast cancer (TNBC), však tyto klinicky stanovené biomarkery postrádá. To znamená, že hormonální terapie nebo cílené léky nepřicházejí v úvahu, takže je na výběr méně možností léčby. Aby bylo možné vyvinout nové léky na míru, je zásadní pochopení molekulárního základu onemocnění. V poslední době se mnoho studií zaměřuje na hledání biomarkerů na úrovni proteinů pomocí proteomiky. Proteiny, zejména jejich post-translační modifikace (PTM), jsou jádrem mnoha buněčných událostí a jejich odhalení může pomoci při pochopení mechanismů rakoviny prsu. Pro objevení molekulárních rysů TNBC, je cílem této studie porovnat proteomická data neléčených rakovinných buněčných linií s buňkami, které podstoupily retinoidní terapii. Důraz bude kladen na PTM, zejména glykosylaci a fosforylaci, Vimentinu a CD44, které byly navrženy jako potenciální biomarkery TNBC v předchozích studiích. Proteinová separace bude provedena pomocí 1D a 2D gelové elektroforézy nebo pomocí SEC-HPLC. Vzorky budou také podrobeny enzymatickému štěpení před identifikací pomocí MALDI-TOF hmotnostní spektrometrie. V případě fosfoproteinového selektivního záchytu bude obohacení provedeno afinitní chromatografií s použitím hrotů pro obohacení fosfopeptidu TiO2 (TopTip). Glykosylované proteiny budou obohaceny pomocí WGA lektinové afinitní chromatografie. Proteiny s významnými rozdíly v PTM mezi ošetřenými a neošetřenými buňkami budou blíže hodnoceny pomocí proteinových databází (MASCOT, STRING a další). Data získaná ze studie budou případně použita k navržení potenciálních biomarkerů pro TNBC.
|
|
|
Proteomic approach for the study of cancer cell line profiles.
Predná, Nikola ; Langová, Denisa (referee) ; Strouhalová,, Dana (advisor)
Triple-negativní karcinom prsu (TNBC), velice agresivní podtyp rakoviny prsu, je známý svou nepříznivou prognózou a omezenými možnostmi léčby. V tuto chvíli je chemoterapie považována za hlavní způsob léčby. Za účelem vyvinutí nových účinných léčiv je snaha pochopit molekulární základ této nemoci. V důsledku toho bylo již několik potenciálně aktivních látek pro tento konkrétní typ rakoviny prsu podrobeno výzkumu. V poslední době se mnoho studií zabývající touto záležitostí provádí za použití proteomiky jakožto prostředku ke studiu proteomů rakovinných buněk. Rakovinové buňky obsahují klíčové rozdíly v proteinech, které regulují mechanismy buňky. Mapování těchto mechanismů může nakonec umožnit diagnostikovat stav organismu. Tato práce se zaměřuje na proteomické studium buněk TNBC a porovnává neošetřené buňky s buňkami, které byly podrobeny léčbě retinoidy. Separace proteinů a peptidů byla úspěšně provedena elektroforézou na 1D a 2D gelu. Kromě toho byly vzorky podrobeny enzymatickému štěpení vybraných proteinů, které byly poté identifikovány pomocí hmotnostní spektrometrie MALDI-TOF (MS). Proteiny, které se podílejí na procesu epiteliálně-mezenchymálního přechodu (EMT), byly poté kvantifikovány a porovnány mezi vzorky.
|
|
|
Modern therapy of chondral defects
Neckař, Pavel ; Havlas, Vojtěch (advisor) ; Zeman, Petr (referee) ; Tuček, Michal (referee)
The thesis describes the application of cultured bone marrow stem cells in the therapy of focal chondral defect of the knee joint. In the experimental part of the work, the goal was to quantitatively and qualitatively compare two sampling sites of bone marrow monocytic aspirates, the iliac crest bone and the proximal tibia, in order to determine a suitable cell source for advanced cell therapy. The sample analysis showed that the amount of monocytic cells and the yield of stem cells from the aspirate obtained were significantly higher in the bone marrow from the iliac crest. We did not find significant qualitative differences between the two sources of stem cells. In the clinical part of the work, I present a description of the surgical procedure and the results of a 1-year follow-up of patients after the implantation of cultured stem cells from the bone marrow, under the name BiCure® orthoMSCp (Bioinova, Prague, Czech Republic), fixed on a commercially available 3D scaffold Chondrotissue® (BioTissue AG, Geneva, Switzerland) using coagulated autologous platelet-rich plasma. The primary objective of the clinical study included the evaluation of the short-term and long-term safety of the applied medical product. The secondary objective of the work included the assessment of the effectiveness of the...
|
|
|
In vitro models for studying Syncytin-1-induced fusion of trophoblast cells
Jech, Lukáš ; Trejbalová, Kateřina (advisor) ; Zíková, Martina (referee)
Trophoblast cell fusion is essential for human placenta development. Apart from initiating blastocyst implantation, syncytialization is critical for optimal nutrient and gas exchange between mother and fo- etus. Multicellular syncytia called syncytiotrophoblast covers the surface of the branched structure of chorionic villi, which is in direct contact with maternal blood. Impairment of the syncytialization process leads to insufficient fetal nutrition and severe pregnancy complications. Syncytia formation is induced by the interaction of the surface glycoprotein of retrovital origin, Syncytin-1, with its receptor. Despite the significance of these processes, the details of cell fusion and trophoblast differentiation remain unk- nown. Furthermore, because of its uniqueness, the human placenta cannot be covered by animal models. As a result, research into human placental development, especially Syncytin-1-induced trophoblast cell fusion, is limited to in vitro trophoblast models. These models include primary trophoblast cell cultures and trophoblast cell lines, which can be obtained by immortalizing cell cultures or extracted from trophoblast tumours. Dedifferentiated trophoblast stem cell cultures were also established. The most recent approach, however, involves the direct reprogramming of dermal...
|
|
|
Potential of mesenchymal stem cells and nanoparticles in the treatment of retinal disorders
Kettner, Ondřej ; Heřmánková, Barbora (advisor) ; Filová, Eva (referee)
Retinal degenerative diseases are recognized worldwide as one of the predominant causes underlying irreversible visual impairment and even blindness. The current standard treatment has helped in both the prevention and supportive treatment of several pathologies, especially if they relate to the retina. However, many of these methods are rather invasive in terms of procedure with the subsequent possibility of infection, inflammation or even retinal detachment with the potential risk of vision loss. These restrictions represent a significant cost to patients' quality of life and also have an economic impact on the healthcare system, so new treatment options are being sought. Mesenchymal stem cells (MSCs) are a promising cell therapy tool for the treatment of many previously untreatable defects and diseases. In addition to MSCs, the use of nanoparticles and nanofibrous carriers is currently being studied in the diagnosis and therapy of retinal diseases. Combining the immunomodulatory and regenerative properties of MSCs and nanotechnologies offers a promising therapeutic strategy for a number of diseases. This thesis is therefore devoted to the current state of the use of nanomaterials, MSCs and their combination that may provide new opportunities in the treatment of retinal degenerative diseases.
|
|
|
Mechanisms of Vascularization in Skin Tissue Engineering
Futóová, Terézia ; Brož, Antonín (advisor) ; Šuca, Hubert (referee)
Tissue engineering is a multidisciplinary field dealing with the fabrication of artificial tissue substitutes for regenerative medicine. Current regenerative medicine uses various types of tissue grafts, which have different advantages and disadvantages depending on their origin, such as insufficient amount of replacement tissue when using autologous grafts or immunogenicity of allogeneic or xenogeneic grafts. An alternative could be artificial tissue replacement. Artificial tissue constructs may consist of a non-living matrix and a cellular component. The cellular component may remodel the construct, form a functional part of the construct, or help integrate the construct into the host body. A significant problem in the formation of such replacements is sufficient vascularization. It is essential to keep cells in larger tissue constructs alive. Vascularization can be enhanced by the addition of vascular endothelial cells that can form capillaries independently within the construct. Vascular formation can also be aided by angiogenic growth factors by their direct application to the construct or by their formation, e.g. in stem cells cultured in the construct. Another approach is 3D bioprinting, allowing direct placement of specific cell types, growth factors or biomaterials in the construct. This...
|
|
|
Awareness of pregnant women about cord blood donation
KOLMANOVÁ, Michaela
This bachelor's thesis is dedicated to donating umbilical cord blood. The thesis consists of a theoretical and practical part. The theoretical part contains a description of the placenta and its functions, umbilical cord blood, stem cells. Furthermore, the theoretical part contains a description of the technique of cord blood collection, contraindications of the collection and processing of umbilical cord blood. It also describes the education of midwives on this issue. Umbilical cord blood banks and legislative provisions are listed here. The aim of the bachelor's thesis was to find out how knowledgeable pregnant women are about this issue. A quantitative research survey was used in the practical part of this work. The questionnaire created contained 18 closed questions. The research team was pregnant women. This set consisted of 244 respondents. Of these, 155 (64%) first-time parents 89 (36%) multi-parents. Of these, 41.39% of respondents know the use of umbilical cord blood. Only 18.03% of respondents know what they need to do to be able to take umbilical cord blood. 19.67% of respondents reported the correct time of collection of umbilical cord blood. 13.52% of respondents reported that the maximum storage period of donated umbilical cord blood is 20 years. Two hypotheses were established in this work. The first hypothesis focused on whether pregnant women are given more information about this issue by a midwife or gynecologist. 5.70% of respondents received information from a midwife. 3.70% of respondents received information from a gynecologist. Statistical processing of the hypothesis has shown us that there is no difference in who gives information. Women are informed from the midwife as well as from the gynecologist. The second hypothesis was whether multi-parents are more informed about umbilical cord blood donation than first-time parents. Multi-parents accounted for 36%, and first-time parents 64%. 46.06% of multi-parents and 38.71% of first-time parents said they knew the use of umbilical cord blood. 20.22% of multi-parents and 19.35% of first-time parents know the time of collection of umbilical cord blood. 50.56% of multi-parents and 39.35% of first-time parents reported that the donation was not risky for newborns. We confirmed the second hypothesis. The results of this work show that women have very little information on the issue of umbilical cord blood donation. The midwife is a competent person, so women could be more education about this issue. This work can be used in seminars for midwives.
|
guest ::
