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Study of rose hip bioactive compounds
Konderla, Patrik ; Skoumalová, Petra (referee) ; Veselá, Mária (advisor)
Purpose of this thesis was research the content of biologically active substances in herbal rosehip extracts and also determination of antimicrobial and antioxidant activity of these extracts. Teoretical part describes informations about Rosa canina L., detailed specification of biologically active substances, specification of antioxidation and overview of methods determine antioxidant compound and antimicrobial activity. In the practical part was by combination of different solvents, procedures and herbal materials, made 21 extracts of Rosa canina L.. From measuring of these extracts, was made extraction curves and also was determined antimicrobial efficiency against bacterial strains Serratia marcescesns, Bacillus subtilis, Bacillus cereus and Micrococcus luteus. Results shows, that also all extracts has some antimicrobial activity, but only against microorganisms Micrococcus luteus and Bacillus cereus. Also was found that bud extracts contains more bioactive compounds soluble in ethanol, meanwhile rose hip extracts contains more bioactive compounds soluble in water.
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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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Encapsulation of plant extracts containing phenolic compounds into nanoparticles and nanofibers
Petrželková, Markéta ; Bokrová, Jitka (referee) ; Skoumalová, Petra (advisor)
The presented bachelor thesis was focused on the encapsulation of extracts containing phenolic substances in order to apply these materials in cosmetics. At work were prepared aqueous, ethanol and lipid extracts of coffee and cocoa. All extracts were characterized on the content of all polyphenols and flavonoids. Their antioxidant aktivity and SPF were also determined. Then the selected extracts were encapsulated into liposomes and polyhydroxybutyrate nanofibres. The basic characteristics of the prepared liposomes were also observed and also their SPF and antioxidant aktivity were monitored. The highest antioxidant aktivity and SPF had liposome particles containing ethanol cocoa extracts. The liposome particles were applied to cosmetic emulsion. These creams evince high antioxidant acitivity and stability determined by analytical centrifugation. Also the prepared nanofibres had high antioxidant aktivity and confirmed the gradual release of the active substances. In conclusion the test cytotoxicity on human keratinocytes confirmed the safety of prepared nanomaterials using the MTT test, which are therefore suitable for cosmetic applications.
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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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Plant-based cosmetic products
Horváthová, Iveta ; Pavelková, Renata (referee) ; Skoumalová, Petra (advisor)
The aim of this bachelor thesis was studying an antimicrobial activity of selected herbal extracts and their encapsulated forms as liposomes. Furthemore, the thesis focuses on determonation of biologically active compounds, natural pigments and antioxidant activity. 6 kinds of herbs were studied - Humulus lupulus, Matricaria chamomilla, Taraxum officinale, Bellis Perennis, Cymbopogon citratus and Rubus Fruticosus. In teoretical part, the general characteristics of herbs and their use are described, it also includes the description of active compounds, microorganisms and used methods. Practical part is focused on characterisation of herbs from the perspective of antioxidant activity, determination of totaly phenolic content and flavonoids, chlorophyll and carotenoids. For analysis of antioxidant activity, concentration of phenolic compounds and flavonoids and antimicrobial activity aqueous and 20% ethanol extracts were prepared. Based on the obtained results, selected extracts were encapsulated into various types of liposomes consisting egg yolk, soy or sunflower lecithin. Generally, ethanol extracts showed higher content of active compounds. For chlorophyll and total carotenoid content determination the highest content of natural pigments was found in Taraxum officinale. Furthemore, antimicrobial activity of extracts and liposomes against 6 strains were determined - Micrococcus luteus, Serratia marcescens, Escherichia coli, Propionibacterium acnes, Staphylococcus epidermidis and Candida glabrata. The highest inhibitory effect was found in Matricaria chamomilla and Cymbopogon citratus samples. Encapsulation effectivity, size, stability and long-term stability of liposomes after a week and two months were determined. From the perspective of zeta potential all the particles were stable and reached a good encapsulation effectivity. It turned out that after two monts of storage the particles decomposed and active compounds into surroundings were released. Selected herbs generally had high content of antioxidants and active compouds. The most suitable lecithin for encapsulation was shown to be sunflower lecithin.
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Use of plasma jet for biomedical application
Doubravová, Anna ; Márová, Ivana (referee) ; Skoumalová, Petra (advisor)
This master´s thesis is focused on the utilization of the sterilization effects of low temperature plasma towards the bacterial microorganisms that occur mainly on the human skin. The plasma sterilization process is fast efficient, non-toxic, environmentally friendly, cost-effective and safe for the operating staff as well as for the patient. Another advantage of using low temperature plasma is to support cell proliferation and wound healing. By combining these advantages, an effective method can be obtained, which would sterilize the wounds sparingly with regard to the surrounding healthy tissue and support the regeneration of the damaged tissue at the same time. In the experimental part, gram positive and gram negative bacteria were used to prove the sterilization effects with respect to different cell wall structure. Staphylococcus epidermidis and Propionibacterium acnes, which cause purulent skin inflammations, were used as gram-positive microorganisms. Serratia marcescens and Escherichia coli were selected from gram positive bacteria. These model organisms were inoculated at various concentrations on culture broths and treated by plasma at a distance of 1 mm from the agar surface. The microwave discharge was generated in argon at a power of 9 W, a gas flow rate of 5 l / min and water cooling to avoid thermal effects on the treated surface. Subsequently, model skin cells of HaCaT were exposed to low temperature plasma and tested for plasma cytotoxicity to demonstrate its healing effects. The obtained results make it possible to state that the sterilizing effects of low-temperature plasma in all tested gram-positive and gram-negative bacterial strains are verified in this work. Finally, tests were demonstrated using a suitable method of the treatment on human skin cells, where the safety and usefulness of the tested low-temperature plasma was demonstrated when applied to shorten the healing process.
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Using different methods of DNA isolation of lactic acid bacteria in molecular biological methods
Chvalkovská, Eva ; Skoumalová, Petra (referee) ; Brázda, Václav (advisor)
This thesis focused on the probiotic bacteria, DNA isolated from these bacteria by three different methods and the effect of isolation on DNA identification using molecular biological methods. Probiotic bacteria are an important part of human intestinal tract. They have an important role in the function of the immune system due to adhesion to the mucosa of the intestinal flora. They create a inhostile environment for pathogens. Probiotic bacteria are commonly taken in the food like dairy products or food supplements. However, overuse of antibiotics is at risk of passing on the intrinsic resistance that probiotic bacteria have to the pathogenic bacteria. The intrinsic resistence they have to maintain the natural homeostasis of the intestinal tract. It is important to effectively identify risky probiotic bacteria that have the ability to transmit resistance to eliminate their presence in food and dietary supplements. Three methods of DNA isolation like phenol extraction method, magnetic particle isolation and commercial kit isolation were used in the experimental part. DNA was isolated from three dietary supplements, namely Biopron 9 premium, Linex forte and GS Lactobacily forte 21. The purity and concentration of the isolated DNA was detected spectrophotometrically. The presence of individual DNA strains in dietary supplements was confirmed by real-time polymerase chain reaction. The best method of isolation in terms of purity and concentration of isolated DNA was evaluated by RT-PCR and spectrophotometry using a commercial kit isolation method.
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Application of natural antibiotics components in hygienic gels
Kubišová, Veronika ; Pavelková, Renata (referee) ; Skoumalová, Petra (advisor)
This bachelor thesis was focused on the preparation and characterization of gels containing a natural component exhibiting an antifungal effect. A part of this work was the research, which was focused on the description of superficial and systemic fungal diseases, antifungal drugs and plant products with a potential antifungal effect. Moreover, biologically active substances found in selected plants was described. Liposome particles that were part of the experimental part were also described and characterized. Considering the subject of this thesis was the preparation of antifungal gel, the basic characteristics of gels such as their drug delivery systems were described. Descriptions of basic methods for the determination of active substances and antifungal activity were not omitted. In the experimental part, three types of extracts (aqueous, ethanolic and lipidic) were prepared from selected plants, which were subsequntly characterized. Also, all kinds of prepared extracts were encapsulated into liposome particles, in which encapsulation efficiency, size and stability by electrokinetic potential were subsequently determined. Prepared extracts in free and encapsulated form were tested for antifungal activity, when testing was performed on Candida glabrata yeast. Moreover, the antimycotic effect of these natural substances was compared with a commercially available antifungal drug, where the drug was also tested in free and encapsulated form. Extracts in which higher active compound content and higher percent inhibitory activity of C. glabrata cells were determined were selected for incorporation into the prepared gels. The result of this work are natural-based gels that could be used to treat infectious fungal diseases. An example of a prepared gel is a gel containing a lipidic extract of marigold or containing liposomes prepared from a lipidic extract of elm pincushion.
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Isolation, characterization and application of active components of prickly pear and aloe vera
Kocúrová, Petra ; Vysoká, Marie (referee) ; Skoumalová, Petra (advisor)
The main goal of this bachelor thesis was the study of antioxidant and antibacterial effect of natural plant extracts, specifically from pulp and peel of leaf of Aloe vera and Opuntia ficus–indica. Liposomes with encapsulated active compounds were prepared and their use in food and cosmetics industry was analyzed. The theoretical part was focused on botanical with and phytochemical description of natural plant and their use in nutrition and medical practice. Also encapsulation and methods used to analyze encapsulated substances were described. In the experimental part basic characteristics of extracts from two kinds of plantusing ethanol as a solvent in its different concentration (0 %, 20 %, 40 %, 80 %) were analyzed. Mainly totalcontent of phenolics, flavonoids and total antioxidant activity were determined. Selected extracts were also encapsulated into liposomes and the encapsulation efficienty, size, stability and long–term stability of liposomes were determined. Antibacterial activity was tested against Grampositive Micrococcus luteus, Staphylococcus epidermidis and Propionibacterium acnes and Gramnegative Serratia marcescems a Escherichia coli. Moreover, antibacterial acvivity against yeast Candida glabrata was tested. Finally, 2 ethanol extracts of plant samples were tested in terms of cytotoxicity by using cell line Caco–2 and profile of selected phenolic compounds ere determined in the selected extract samples by using high performance liqiud chromatography. 20% ethanol solution was chosen as the most efficient extract solvent which contained the highest concentration of total phenolic content and concurrently all tested extracts showed antioxidant activity. However, there was minimal or none antibacterial effect. Prepared liposomes from extracts were stable. Also product for cosmetics and food industry was designed, specifically healing gel with moisturizing effect and paste made of liposomes as food supplement for athletes.
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Preparation and characterization of nanofiber wound dressings
Jiroušková, Pavla ; Uhlířová, Renata (referee) ; Skoumalová, Petra (advisor)
The presented bachelor thesis deals with the preparation and characterization of nanofiber wound dressings. The materials used for the preparation of nanofibers were poly(3-hydroxybutyrate) and gelatin. Active substances, such as ampicillin and ibuprofen, were incorporated into the wound dressings. In the theoretical part of the work skin, wound healing, and nanofibers and their properties were described. This part also deals with the use of nanofibers as wound dressings. Additionally, it depicts materials suitable for the preparation of nanofiber wound dressings. Furthermore, this section contains a review focused on various methods of preparation of nanofibers, drugs that can be incorporated into the fibers and the subsequent determination of the safety of the prepared dressings. In the experimental part, nanofiber wound dressings containing ampicillin and ibuprofen were produced. The maximum possible amount of active substance feasible to be incorporated into the nanofibers has been determined. Subsequently, combined wound dressings were prepared to study the release rate of active substances into the model environments. The release in the time intervals was determined spectrophotometrically. Finally, antimicrobial activity assays were performed to test the effect of dressings containing ampicillin on selected strains of microbiirganism: S. Epidermidis, E. Coli and M. Luteus
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