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Hydrogel Scaffolds for Tissue Engineering
Chocholatá, Petra ; Babuška, Václav (advisor) ; Kovařík, Tomáš (referee) ; Pešta, Martin (referee)
Bone tissue engineering attempts to mimic the natural behavior of hard connective tissues. Currently, there is an active research and development of new functional materials in this field. The main direction in the treatment of bone tissue defects is the use of biodegradable composite materials based on biopolymers enriched with an inorganic component, possibly in combination with a cellular component. In general, hydrogels play an important role in tissue engineering due to their properties, as they can respond relatively well to changes in environmental conditions and are used as a suitable medium for cells to grow and proliferate. Structure of hydrogels also allows nutrients and waste products to diffuse, and the last but not least, thanks to this they can also serve as drug carries. A major advantage of hydrogels is that they can be injected with subsequent in situ gelation. In this work, physically cross-linked hydrogels based on polyvinyl alcohol (PVA) and hyaluronic acid (HA) in different volume ratios enriched with hydroxyapatite (HAp), an inorganic component inherent to bone tissue, were compared. Incorporation of HAp into the organic matrix was performed in two different ways, by in situ synthesis directly in the PVA/HA matrix and by physical mixing of pre-prepared HAp. The composition of...
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The Role of MicroRNAs in the Progression of Cardiovascular Diseases
Černá, Václava ; Pešta, Martin (advisor) ; Vítovec, Jiří (referee) ; Chottová Dvořáková, Magdaléna (referee)
Current applied research of microRNA molecules is focused on the identification of their roles in the pathogenesis of various types of diseases, including cardiovascular (CV), with the assumption of their use as biomarkers or treatment targets. Cardiovascular diseases have a high incidence and prevalence in the population, are predominantly chronic, and are also a leading cause of hospitalization and death, and therefore represent a major burden on patients and the health system. Many metabolic parameters are currently used in the diagnosis of cardiovascular diseases and cardiovascular risk assessment, mostly proteins and lipids, while parameters from the group of nucleic acids, whether in the form of mutations in DNA molecules, or various RNA molecules, are very limited so far. Despite the undeniable importance and contribution of metabolic biomarkers in the diagnosis and prognosis of cardiovascular diseases, these markers still do not allow effective identification of all patients at high risk, and therefore additional molecules are being sought that could contribute to the accuracy of this identification. Finding other potential diagnostic and prognostic markers to contribute to the more precise CV risk assessment, and subsequently also to the more effective treatment of patients at high risk,...
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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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Prognostic and Predictive Factors in Patients with Metastatic Colorectal Cancer Treated with Targeted Therapy
Šorejs, Ondřej ; Fiala, Ondřej (advisor) ; Pešta, Martin (referee) ; Poprach, Alexandr (referee)
Introduction: In recent years, significant progress has been made in the treatment of metastatic colorectal cancer (mCRC) due to the use of targeted therapy. Bevacizumab is a monoclonal antibody (mAb) against vascular endothelial growth factor (VEGF) and cetuximab and panitumumab are monoclonal antibodies against epidermal growth factor receptor (EGFR). MicroRNAs (miRs) are single-stranded non-coding RNA chains of 18-25 nucleotides in length that are involved in the regulation of gene expression and thus significantly influence the biological behaviour of cancer cells. Dysregulations at the miR level are considered promising candidates for prognostic and predictive biomarkers. Aims: The aim of this work, divided into two parts, was to evaluate the association of dysregulation of selected miRs with the effect of targeted therapy in patients with mCRC. Methods: The first study included 63 mCRC patients treated with chemotherapy and bevacizumab. The second study included 46 mCRC patients treated with chemotherapy and cetuximab or panitumumab. Assessment of treatment effect included progression-free survival (PFS), overall survival (OS), objective response rate (ORR) and disease control rate (DCR). Samples of frozen tumor and adjacent non- tumor tissue obtained during primary tumor surgery were used...
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Prediction of Treatment Effect in Solid Tumors
Houfková, Kateřina ; Pešta, Martin (advisor) ; Fiala, Ondřej (referee) ; Šána, Jiří (referee)
Current research of treatment of oncological diseases is focused on the identification of changes that occur in the process of carcinogenesis and on the search for therapeutics that would make it possible to eliminate the manifestation of these changes. Early diagnosis, estimating prognosis, monitoring of the course of the disease and prediction of the effect of treatment are increasingly associated with the determination of appropriate biomarkers. The assessment of predictive biomarkers should become an integral part of treatment decisions and, as a result, help patients to prolong life and improve quality of life. The aim of my thesis was to find prognostic and predictive biomarkers for more effective treatment of cancer patients. In cooperation with a number of clinical departments of the University Hospital in Pilsen, I focused on the research of these biomarkers at the Department of Biology, especially in non-small cell lung cancer, endometrial cancer, colorectal cancer, liver metastases of colorectal cancer, clear cell renal cell carcinoma and sarcomas. We focused on the research of molecules that are involved in the process of carcinogenesis and whose levels in body fluids or directly in tumor tissue would be able to predict further disease development or therapeutic response. Using molecular...
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Assessment of Biological Activity of Colorectal Carcinoma using RT REAL-TIME PCR
Pešta, Martin ; Topolčan, Ondřej (advisor) ; Pešek, Miloš (referee) ; Pecen, Ladislav (referee)
The author in his doctoral thesis, proposed primers and conditions optimized design Quantitative PCR for determining the gene expression of MMP-7, TIMP-1, MMP-2 and TIMP- second Revealed the presence of gene expression of GAPDH TIMP-1 and TIMP-2 in cancer line HT-29 SW480 and SW620. Noted the high level expression of MMP-7 in line HT-29th Expression of the 2-3 orders of magnitude higher than the lines SW480 and SW620. MRNA expression of MMP-2 in line HT-29 noticed. MRNA expression of MMP-2 by the detected lines SW480 and SW620. He found that determining absolute and relative expression in tumor lines is ekvivalentní. significantly higher mRNA expression of MMP-7, TIMP-1, MMP-2 and TIMP-2 in tumor tissue compared to normal tissue. It can be used in therapy. Noticed the presence of correlation of gene expression MMP-7, TIMP-1, MMP-2 and TIMP-2 with survival and DFI. He showed that a higher stage cancer correlated with a higher median momery MMP-2/TIMP-2. 7, Noticed the difference in gene expression between TS, TP and DPD in control and tumor tissue. Scored marginally significant increase in TS expression in colon tumors compared to tumors rectosigmoid and rectum. This finding can be used in treatment decisions. scored marginally significant correlation between the expression of TS and DPD. This finding can be...
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Epigenetic Alternations in Head and Neck Squamous Cell Carcinoma: Potential Biomarkers
Kovaříková, Helena ; Palička, Vladimír (advisor) ; Friedecký, David (referee) ; Pešta, Martin (referee)
5 1 Summary ENG Head and neck squamous cell carcinomas are a group of heterogenic tumors arising from epithelial tissue of aerodigestive tract characterized by difficult diagnosis, treatment and prognosis. Alcohol consumption, smoking and high-risk human papillomavirus infection are very well described risk factors of head and neck cancer development. Sinonasal carcinomas are group of malignancies developing in nasal and paranasal sinuses. Oropharyngeal carcinomas are malignancies developing in the throat area downstream to oral cavity. microRNAs (miRNAs) are short (~23 nucleotides) non-coding RNA molecules participating in regulation of gene expression. Primary function of miRNAs is negative translation regulation as part of RISC (RNA-induced silencing complex) by translational repression and mRNA degradation. miRNAs are involved in various disease pathologies such as neurodegenerative diseases, metabolic disorders and cancer. Moreover, microRNAs have been recognized as key molecules in cancer development and progression in various types of tumors. My doctoral study research activities were focused on investigation of relative expression of several preselected miRNAs (using real-time PCR) and the relationship between their expression and clinicopathological characteristics of the squamous cell carcinoma...
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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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