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Study of the effect of zinc oxide nanoparticles on cell migration
Kutějová, Monika ; Zumberg, Inna (referee) ; Chmelíková, Larisa (advisor)
Zinc oxide nanoparticles (ZnO NPs) are widely used across various industries due to their properties, and with the development of nanomaterials, they are also finding applications in medicine. This bachelor’s thesis investigates the cytotoxic effects of these nanoparticles on cultured cell lines. The study focused on the human osteosarcoma cell line Saos-2 and the fibroblast cell line NIH-3T3. During in vitro experiments, the cells were exposed to different concentrations of ZnO NPs. The cytotoxicity of each concentration was assessed using the MTT assay, which provided information on cell viability. Based on the MTT assay results, appropriate concentrations of ZnO NPs were selected for conducting a Scratch assay, with the highest concentration corresponding to the determined inhibitory concentration for NIH-3T3. The cytotoxic effects were thus evaluated by studying cell migration. Subsequent statistical analysis determined the extent of cytotoxic effects of ZnO NPs depending on the concentration and cell type.
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Interaction of yeast and microalgae extracts with cell cultures
Serikbay, Adilkhan ; Holub, Jiří (referee) ; Márová, Ivana (advisor)
The objective of this bachelor thesis was to optimize the extracting of bioactive compounds from microorganisms for the purpose of characterizing and evaluating their cytotoxicity on human cell line HaCaT. During the characterization phase, extract samples were analyzed using spectrophotometric techniques. After exposing the cell line to selected extracts, a cytotoxicity test was performed to assess cell viability using MTT analysis. The theoretical part encompassed an extensive literature review focused on cultivation techniques, human cell lines, cytotoxicity analysis, and viability assessments. The practical section provided a detailed exploration of contemporary extraction techniques utilizing a wide range of both polar and non-polar solvents to enhance the efficiency of extracting active compounds from the yeast strain Rhodotorula toruloides and microalgae Chlorella vulgaris. Active compounds such as ubiquinone, ergosterol, lycopene, and torularhodin were identified in the yeast, while the microalgae were found to contain lutein, chlorophylls, as confirmed by HPLC analysis. The biocompatibility of the optimized extracts from selected strains was verified using the MTT assay on HaCaT cell lines. The aqueous extract from microalgae Chlorella vulgaris exhibited lower toxicity to HaCat cell line, with viability rates exceeding 85%. Yeast extracts from Rhodotorula toruloides initially showed higher viability rates, which, however, decreased with increasing extract concentrations.
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Study of biological effects of some plant materials
Drabíková, Nela ; Byrtusová, Dana (referee) ; Márová, Ivana (advisor)
The presented bachelor thesis deals with the preparation of aqueous, ethanolic and hexane extracts obtained from plant sources with increased content of oil, their characterization and cytotoxicity testing. In the theoretical part, a review focused on basic information about the used plants and their effects on the human body was elaborated. Furthermore, the active ingredients of the extracts, methods of their determination and the principle of the cytotoxicity test were described. In practical part the content of polyphenols and flavonoids was determined spectrophotometrically. Further, total antioxidant activity was measured. The fatty acids composition and the percentage of individual fatty acids were determined by gas chromatography. Next, MTT cytotoxicity assay was performed using human cell culture to confirm the safety of extracts for potential use in food, cosmetic and pharmaceutical industries.
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Use of cell cultures to test cytotoxicity of natural substances
Blažková, Veronika ; Skoumalová, Petra (referee) ; Márová, Ivana (advisor)
This thesis is aimed on study of cell cultures, primarily on introduction of techniques for cultivation of human cells and optimization of their usage in cytotoxicity tests. The thesis is aimed preferably on natural substances and liposomes that are suitable to be used in cosmetics. Theoretical part contains literature review focused on overview of cell cultures, appropriate cultivation conditions for in vitro testing and summary of cytotoxicity tests. Experimental part contains characteristics of natural extracts with antimicrobial effect in terms of content of polyphenols, flavonoids and antioxidant activity. Further, these extracts were encapsulated into the liposomes. Encapsulation effectivity was determined. Distribution of particles size by dynamic light scattering and finally also stability of prepared particles were also determined. Long-term stability in water and gel environment was tested too. This thesis also contains study of optimization of cell culture of human skin keratinocytes cultivation and use of those cells for testing cytotoxicity of selected natural substances by MTT method.
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Testing the effects of a mixture of beta-glucans and antimicrobial extracts
Mihálová, Veronika ; Bendová, Agáta (referee) ; Němcová, Andrea (advisor)
The bachelor thesis aims to describe the effects of a mixture of selected natural substances in conjunction with -glucan obtained from the cell wall of the studied yeast species. The theoretical part describes bioactive substances and compounds with antioxidant activity. Subsequently, selected species of plants and herbs are characterized as direct sources of bioactive substances. In parallel, the following chapters of the theoretical part present as sources of -glucan representatives of yeast species with their morphological characterization. - glucan, its structure, biological effects, as well as the principles of methods used in the determination and preparation of bioactive mixtures of plant extracts and -glucan are described. In the experimental part of the work, attention is paid to the preparation, cultivation of yeast, isolation, and determination of -glucan content using the enzymatic method. The amount of polyphenols, flavonoids, and antioxidant activity in aqueous plant extracts is determined. The content of fatty acids, carotenoids, and coenzyme Q is determined in yeast. Subsequently, the antimicrobial and antioxidant effects of mixtures of plant extracts and -glucan are tested.
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Testing of the cytotoxicity of potential drugs in the cell lines together with the measurement of their passage through the blood brain barrier by the PAMPA method
Šinaľová, Lucia ; Hofman, Jakub (advisor) ; Jansová, Hana (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacology & Toxicology Student: Lucia Šinaľová Supervisor: RNDr. Jakub Hofman, Ph.D. Title of diploma thesis: Testing of the cytotoxicity of potential drugs in the cell lines together with the measurement of their passage through the blood brain barrier by the PAMPA method This master thesis focuses on the prediction of tacrine derivatives permeability through blood- brain barrier and their cytotoxicity. We studied tacrine-benzothiazole and tacrine- thiaquinazoline derivates as potential drugs for treatment of Alzheimer's disease. The reason of testing new tacrine derivates was effort to find substances with same effect and lower toxicity, for which was the tacrine discarded from clinical practice. The probable permeability of the studied substances was determined in vitro by the PAMPA method. Based on the permeation coeficient values we have identified that tacrine-benzothiazole derivates A-D (Pe= 8,31- 16,9×10-6 cm/s) and tacrine-thiaquinazoline derivates 1, 3, 4 (Pe= 8,59-14,9×10-6 cm/s) were permeable through blood-brain barrier, tacrine-thiaquinazoline derivate 2 (Pe= 3,79×10-6 cm/s) had uncertain permeability and tacrine-thiaquinazoline derivate 5 (Pe= 2,0×10-6 cm/s) was not permeable. In the pre-clinical evaluation of...
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Testing the effects of a mixture of beta-glucans and antimicrobial extracts
Mihálová, Veronika ; Bendová, Agáta (referee) ; Němcová, Andrea (advisor)
The bachelor thesis aims to describe the effects of a mixture of selected natural substances in conjunction with -glucan obtained from the cell wall of the studied yeast species. The theoretical part describes bioactive substances and compounds with antioxidant activity. Subsequently, selected species of plants and herbs are characterized as direct sources of bioactive substances. In parallel, the following chapters of the theoretical part present as sources of -glucan representatives of yeast species with their morphological characterization. - glucan, its structure, biological effects, as well as the principles of methods used in the determination and preparation of bioactive mixtures of plant extracts and -glucan are described. In the experimental part of the work, attention is paid to the preparation, cultivation of yeast, isolation, and determination of -glucan content using the enzymatic method. The amount of polyphenols, flavonoids, and antioxidant activity in aqueous plant extracts is determined. The content of fatty acids, carotenoids, and coenzyme Q is determined in yeast. Subsequently, the antimicrobial and antioxidant effects of mixtures of plant extracts and -glucan are tested.
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Study of biological effects of some plant materials
Drabíková, Nela ; Byrtusová, Dana (referee) ; Márová, Ivana (advisor)
The presented bachelor thesis deals with the preparation of aqueous, ethanolic and hexane extracts obtained from plant sources with increased content of oil, their characterization and cytotoxicity testing. In the theoretical part, a review focused on basic information about the used plants and their effects on the human body was elaborated. Furthermore, the active ingredients of the extracts, methods of their determination and the principle of the cytotoxicity test were described. In practical part the content of polyphenols and flavonoids was determined spectrophotometrically. Further, total antioxidant activity was measured. The fatty acids composition and the percentage of individual fatty acids were determined by gas chromatography. Next, MTT cytotoxicity assay was performed using human cell culture to confirm the safety of extracts for potential use in food, cosmetic and pharmaceutical industries.
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Testing of the cytotoxicity of potential drugs in the cell lines together with the measurement of their passage through the blood brain barrier by the PAMPA method
Šinaľová, Lucia ; Hofman, Jakub (advisor) ; Jansová, Hana (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacology & Toxicology Student: Lucia Šinaľová Supervisor: RNDr. Jakub Hofman, Ph.D. Title of diploma thesis: Testing of the cytotoxicity of potential drugs in the cell lines together with the measurement of their passage through the blood brain barrier by the PAMPA method This master thesis focuses on the prediction of tacrine derivatives permeability through blood- brain barrier and their cytotoxicity. We studied tacrine-benzothiazole and tacrine- thiaquinazoline derivates as potential drugs for treatment of Alzheimer's disease. The reason of testing new tacrine derivates was effort to find substances with same effect and lower toxicity, for which was the tacrine discarded from clinical practice. The probable permeability of the studied substances was determined in vitro by the PAMPA method. Based on the permeation coeficient values we have identified that tacrine-benzothiazole derivates A-D (Pe= 8,31- 16,9×10-6 cm/s) and tacrine-thiaquinazoline derivates 1, 3, 4 (Pe= 8,59-14,9×10-6 cm/s) were permeable through blood-brain barrier, tacrine-thiaquinazoline derivate 2 (Pe= 3,79×10-6 cm/s) had uncertain permeability and tacrine-thiaquinazoline derivate 5 (Pe= 2,0×10-6 cm/s) was not permeable. In the pre-clinical evaluation of...
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