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Catecholamine modified polymer-composite bone cement: preparation, characterization and in-vitro evaluation
Raszková, Alena ; Vojtová, Lucy (referee) ; Michlovská, Lenka (advisor)
Tato bakalářská práce je zaměřena na přípravu, charakterizaci a in-vitro hodnocení cytotoxicity katecholaminem modifikovaných fosfátových cementů. Teoretická část práce se zabývá charakteristikou cementů na bázi fosforečnanu vápenatého a jejich materiálových vlastností. Obsahuje také přehled aditiv používaných pro modifikaci cementů za účelem zlepšit jak jejich chemofyzikální, tak biologické vlastnosti. Rovněž je zde diskutováno, dnes stále častější, využití katecholaminu v tkáňovém inženýrství. První část experimentální práce se zaměřuje zejména na samotnou optimalizaci přípravy a charakterizaci katecholaminem modifikovaný fosfátový cement. Kinetika fázového složení byla hodnocena pomocí rentgenové difrakční analýzy a chemické složení pomocí infračervené spektroskopie. Druhá část práce je poté zaměřena na optimalizaci a hodnocení in-vitro cytotoxicity modifikovaného cementu. V závěru bylo zjištěno, že modifikace katecholaminem zrychluje míru konverze cementu během vytvrzování. U takto připravených vzorků byl stupeň konverze vypočten na 93 % po 3 dnech vytvrzování, zatímco u nemodifikovaných vzorků dosahoval ve stejný čas pouze 79 %. Pro in-vitro testování byla navržena nová metoda přípravy vzorků. Vzhledem k výsledkům testování je nutná další optimalizace této nové metody a podmínek testů. Díky svým atraktivním vlastnostem se katecholaminem modifikovaný fosfátový cement zdá být zajímavým materiálem pro regeneraci kostí.
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Preparation of bioactive wound dressings and testing their interaction with human cells
Bendová, Agáta ; Márová, Ivana (referee) ; Skoumalová, Petra (advisor)
The thesis was focused on the preparation and optimization of the preparation of wound dressing from materials with bioactive ingredients. In this work were prepared nanofiber dressings based on polyhydroxybutyrate and non-fibrous dressings from alginate and chitosan. Nanofibers were prepared by electrospinning and forcespinning methods. The bioactive components, which were used to functionalize the prepared dressings, were plant extracts, clotrimazole, ampicillin, lysozyme, and proteolytic enzymes. The theoretical part is focused on the description of the use of nanofibrous and non-fibrous materials in medicine, characterization of materials for the production of wound dressings and bioactive components. Furthermore, this section describes the methods used to prepare and characterize wound dressings. In the practical part were prepared aqueous and oil extracts from selected plants. Extracts were characterized for polyphenols content and antioxidant activity. PHB-based nanofibers were prepared using electrospinning and forcepinning methods. Nanofibers were enriched with selected plant oil extracts and clotrimazol. Modified nanofibres were detemined for antioxidant activity, short-term and long-term stability. Non-fibrous wound dressings were prepared from alginate and chitosan. These dressings were functionalized by the addition of selected aqueous extracts, ampicillin, lysozyme, papain, bromelain, and collagenase. Non-fibrous wound dressings were determined for antioxidant activity, short-term stability and proteolytic activity. The prepared wound dressings were tested for their antimicrobial effects on cultures of Micrococcus luteus, Serratia marcescens, Staphylococcus epidermidis and Escherichia coli. In conclusion, successfully prepared bioactive wound dressings with antioxidant and antimicrobial agents were tested for safety on human cells. The determination was performed using the MTT cytotoxicity test on human keratinocytes.
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Preparation and characterization of modern wound covers
Balášová, Patricie ; Pernicová, Iva (referee) ; Skoumalová, Petra (advisor)
This diploma thesis is focused on the study of bioactive wound dressings. During the thesis, hydrogel, lyophilized and nanofiber wound dressings were prepared. Hydrogel and lyophilized wound dressings were prepared on basis of two polysaccharides – alginate and chitosan. Nanofiber wound dressings were prepared by spinning polyhydroxybutyrate. All prepared wound dressings were enriched with bioactive substances, which represented analgesics (ibuprofen), antibiotics (ampicillin) and enzymes (collagenase). Into hydrogel and lyophilized wound dressings were all the mentioned active substances incorporated, whereas nanofiber wound dressings were only with ibuprofen and ampicillin prepared. The theoretical part deals with the anatomy and function of human skin. There was explained the process of wound healing and also there were introduced available modern wound dressings. The next chapter of the theoretical part deals with materials for preparing wound dressings (alginate, chitosan, polyhydroxybutyrate) and with active substances, which were used during the experimental part of this thesis. In the theoretical part, the methods of preparation of nanofiber wound dressings and also the methods of cytotoxicity testing used in this work were presented. The first part of the experimental part of this thesis was focused on preparing already mentioned wound dressings. Then, their morphological changes over time and also the gradual release of incorporated active substances into the model environment were monitored. The gradual release of ampicillin was monitored not only spectrophotometrically, but also by ultra-high-performance chromatography. In wound dressings, in which collagenase was incorporated, was also the final proteolytic activity of this enzyme monitored. The effect of the active substances was observed on three selected microorganisms: Escherichia coli, Staphylococcus epidermidis and Candida glabrata. The cytotoxic effect of the active substances on the human keratinocyte cell line was monitored by MTT test and LDH test. A test for monitoring the rate of wound healing – a scratch test – was also performed.
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Properties of aminoclay complexes and biologically active substances
Dušek, Jakub ; Pekař, Miloslav (referee) ; Mravec, Filip (advisor)
This paper builds on previous research of aminoclay complexes in undergraduate studies. Theoretical part deals with study of current problems of aminoclay complexes with bioactive substances and the choice of substances for complexing with aminoclay. The experimental part consists of preparation of aminoclay complexes with selected bioactive agents at various concentrations. Verification of binding of bioactive agents to the aminoclay matrix was performed by Elemental Analysis (EA) and Fourier-transform infrared spectroscopy (FT-IR). For finding of the bound amount of bioactive substance were used the Ultra Performance Liquid Chromatography (UPLC®) and analysis of the cytotoxic properties of the formed complexes by used by the MTT assay. The main motivation of this study is to create new complexes with improved characteristics that would replace existing forms of substances used in pharmaceutical and biomedical applications.
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Vliv proteinové korony na toxicitu částic
Jemelíková, Vendula
When foreign particles enter the organism, they come into contact with the physiological environment and subsequent adsorption of naturally occurring biomolecules on the surface of the particles. Protein corona is formed during the adsorption of proteins, which fundamentally changes the physico-chemical properties of the particles. The aim of this work was to focus on the change in properties and toxicity of particles caused by the formation of a protein corona around selected particles. The formation of protein corona around different particles based on hydroxyapatites (CaNPs), iron (FeNPs), platinum (PtNPs) and titanium (TiNPs), was characterized and proved using electrophoretic methods, and subsequently the change in size, zeta potential and cytotoxicity was monitored depending on the formation of a protein corona. Based on the results, it is possible to claim that in all cases particles with a protein corona were less toxic to breast tumor cells, than particles without a corona and CaNPs were the most cytotoxic. In general, it is possible to say that the formation of protein corona around the tested nanoparticles affects their properties and reduces cytotoxicity and oxidative stress.
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Comparison of in vitro methods for the study of cytotoxicity
Eliášová, Pavla ; Maixnerová, Jana (advisor) ; Bárta, Pavel (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacology and Toxicology Student: Pavla Eliášová Supervisor: RNDr. Jana Maixnerová, Ph.D. Title of diploma thesis: Comparison of in vitro methods for the study of cytotoxicity People are exposed to a growing number of toxic substances from the environment. Endocrine disrupting chemicals (EDCs) are a broad category of molecules that are thought to cause adverse effects on the endocrine system by interfering with the synthesis, transport, degradation or action of endogenous ligands. One of the aims of this thesis was to determine the in vitro toxicity of 17 selected endocrine disruptors on the human hepatocellular carcinoma HepG2 cell line. Cell viability was determined using the CellTiter96® AQueous One Solution Cell Proliferation Assay colorimetric method, the principle of which is the reduction of MTS to the colored product formazan by mitochondria in viable cells. The cytotoxic potential of the compounds was expressed by using the toxicological parameter IC50, which was measured in three time intervals (6, 12 and 24 hours). For 14 substances: Atrazine, DHEP, Bisphenol A, Carbofuran, 3-hydroxycarbofuran, Cypermethrin, DDE, DES, MEHP, PCB 118, PCB 153, PFOA, PFOS, Propiconazole, IC50 > 100 µM (respectively > 250 µM) was...
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Study of the expression and toxicity of catechol derivates in MCF-7 cell line
Kleplová, Dominika ; Carazo Fernández, Alejandro (advisor) ; Vokřál, Ivan (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacology and Toxicology Student: Dominika Kleplová Supervisor: PharmDr. Alejandro Carazo, PhD. Title of the Diploma thesis: Study of the expression and toxicity of catechol derivatives in MCF-7 cell line Estrogens perform many important functions in the human body. They act by binding to estrogen receptors (ER) and thus regulate reproduction, the menstrual cycle, bone density, cholesterol metabolism or brain function. They also play an important role in the development and onset of breast cancer, where the amount of ER expressed is used as a very important biomarker in patients suffering from this disease. Despite ongoing research, breast cancer is considered to have the highest mortality rate. Catechols are organic compounds. In the human body, they can occur as metabolites in the degradation of benzene and estrogens or other endogenous compounds such as neurotransmitters and their precursors. Catechols are known to be involved in redox processes in the body, to exert antioxidant and toxic effects, to interfere with protein function and to cause DNA strand breaks. Their positive effect on breast cancer therapy is the subject of research, but it has not yet been fully explored. The subject of this thesis is to study the...
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Cytotoxic activity of selected Narcissus cultivars in vitro.
Maršálková, Aneta ; Hradiská Breiterová, Kateřina (advisor) ; Jošt, Petr (referee)
Charles University, Faculty of Pharmacy in Hradec Králové Department of Pharmacognosy and Pharmaceutical Botany (16-16230) Author: Aneta Maršálková Supervisor: PharmDr. Kateřina Hradiská Breiterová, Ph.D. Title of thesis: Cytotoxic activity of selected Narcissus cultivars in vitro. Key words: Narcissus, Amaryllidaceae alkaloids, cytotoxicity, HepG2 The main focus of this diploma thesis is the screening of summary alkaloidal extracts of selected cultivars of the genus Narcissus for their cytotoxic activity against HepG2 liver carcinoma cell line. In the second phase, the cytotoxic activity of the majorly represented Amaryllidaceae alkaloids (AmA) was screened, followed by an IC50 value determination. All studied alkaloidal extracts underwent also GC-MS and GC-FID qualitative and quantitative analysis to identify as many alkaloids as possible. A total of 23 alkaloids, namely 11,12-didehydroanhydrolycorine, 1-O-acetyl-10-norpluviine, 9-O-methylpseudolycorine, anhydropseudolycorine, assoanine, dehydroassoanine, dihydrolycorine, epi-norgalanthamine, galanthamine, galanthine, haemanthamine, hippeastrine, incartine, caranine, lycoramine, lycoraminone, lycorine, narwedine, norpluviine, pluviine, pseudolycorine, sanguinine, tazettine and several unspecified alkaloids of the homolycorine structural type were...
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Determination of renal toxicity of antineoplastics in vitro
Zádrapová, Marie ; Maixnerová, Jana (advisor) ; Smutná, Lucie (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacology and Toxicology Student: Bc. Marie Zádrapová Supervisor: RNDr. Jana Maixnerová, Ph.D. Title of diploma thesis: Determination of renal toxicity of antineoplastics in vitro BRAF inhibitors are important antineoplastics. They work on the principle of inhibition of certain types of protein kinases and turned out to be very efficient for the treatment of melanoma. One of their disadvantages is relatively early onset of resistance; thus, it is important to look for new combinations of drugs that are already in use or work on the development of new structures with similar inhibition efficacy on melanoma cells. Encorafenib and its combination with binimetinib have been shown to be very promising drugs from the group of BRAF inhibitors, however, potential renal toxicity may be a therapeutic limitation. This thesis was focused on the determination of in vitro cytotoxicity of encorafenib on different types of renal cells in three time intervals and on its comparison with two drug standards - amphotericin B and paracetamol. Three types of morphologically and functionally different kidney cells (PODO / TERT256, HK-2 and HEK293) were used for this purpose. The cytotoxic potential was measured by colorimetric method CellTiter 96®...
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Catecholamine modified polymer-composite bone cement: preparation, characterization and in-vitro evaluation
Raszková, Alena ; Vojtová, Lucy (referee) ; Michlovská, Lenka (advisor)
Tato bakalářská práce je zaměřena na přípravu, charakterizaci a in-vitro hodnocení cytotoxicity katecholaminem modifikovaných fosfátových cementů. Teoretická část práce se zabývá charakteristikou cementů na bázi fosforečnanu vápenatého a jejich materiálových vlastností. Obsahuje také přehled aditiv používaných pro modifikaci cementů za účelem zlepšit jak jejich chemofyzikální, tak biologické vlastnosti. Rovněž je zde diskutováno, dnes stále častější, využití katecholaminu v tkáňovém inženýrství. První část experimentální práce se zaměřuje zejména na samotnou optimalizaci přípravy a charakterizaci katecholaminem modifikovaný fosfátový cement. Kinetika fázového složení byla hodnocena pomocí rentgenové difrakční analýzy a chemické složení pomocí infračervené spektroskopie. Druhá část práce je poté zaměřena na optimalizaci a hodnocení in-vitro cytotoxicity modifikovaného cementu. V závěru bylo zjištěno, že modifikace katecholaminem zrychluje míru konverze cementu během vytvrzování. U takto připravených vzorků byl stupeň konverze vypočten na 93 % po 3 dnech vytvrzování, zatímco u nemodifikovaných vzorků dosahoval ve stejný čas pouze 79 %. Pro in-vitro testování byla navržena nová metoda přípravy vzorků. Vzhledem k výsledkům testování je nutná další optimalizace této nové metody a podmínek testů. Díky svým atraktivním vlastnostem se katecholaminem modifikovaný fosfátový cement zdá být zajímavým materiálem pro regeneraci kostí.
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