|
|
Preparation of nanoparticles and nanofibers with antimicrobial components
Kubišová, Veronika ; Slaninová, Eva (referee) ; Skoumalová, Petra (advisor)
This thesis addresses the problem of inadequate current wound therapy and presents a solution in the form of nanomaterial-based wound dressings (coverings). Specifically, it focuses on the development and characterization of various nanofibrous materials with integrated liposome particles that could serve as a source of therapeutic agents and be useful specifically in the field of a wound therapy. The review in the introductory part of the thesis first focused on the mentioned problem, which describes the shortcomings of existing conventional dressing materials. A description of human skin from an anatomical and functional point of view was not omitted, nor was the wound healing process itself. Different types of covering materials were also presented. However, a large part was focused on nanomaterials and their use in the field of the skin wound therapy. The nanomaterials mentioned were mainly liposome particles and nanofibres, as the experimental part of the work was focused on these structures. The description dealt with the characterization of these structures as well as the biopolymers used for their preparation. However, the aim was to prepare nanofibers with liposome content, so the method of forming such systems was described. Various therapeutic agents are also an integral part of the covering materials, especially those that suppress the development of infection and reduce wound pain; therefore, the search focused on the antibiotic ampicillin and the analgesic ibuprofen. The experimental part was devoted to the actual production of nanofibers with liposomes and also to the qualitative demonstration of the presence of liposomes in electrostatically prepared nanofibers. The selected polymeric components of these systems were polyhydroxybutyrate (PHB) and gelatine. However, the nanofibers and liposome particles (as well as combined liposomes with PHB) were first prepared in their own form and characterized mainly in terms of the gradual release of the drug substances. The results obtained were then compared with those of therapeutic drug delivery via combined nanofibrous structures with liposomes. For these combined structures, the aim was to achieve synergy in drug delivery between these systems. The aim of this work was to create a biomaterial covering with the controlled drug release. The drugs contained in these materials were the aforementioned ampicillin and ibuprofen. An important part of the work was then to determine the safety of the prepared materials which were tested for cytotoxicity, where the tests performed were MTT test and LDH test. And the actual wound healing ability of the nanofibers was then monitored in a scratch test or a "wound healing" test. At the end of the paper, recommendations for future work on this topic are given.
|
|
|
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.
|
|
|
Vliv kovů v potravinářství na bakteriální virulenci
Pavezka, Pavel
The bachelor's thesis deals with the issue of gene transfer of resistance of β-lactam antibiotics by means of horizontal gene transfer, specifically transformation. It deals with the transfer of resistance genes through the food chain, when Escherichia coli can subsequently cause foodborne illness. It also points to the current risks of resistance to β-lactam antibiotics and an alternative method of replacing antibiotics with an iron complex together with an antibiotic. The experimental part is demonstrating the antibacterial effect of the Fe16+AMP complex on the ampicillin-resistant bacterial strain E. coli CCM 4225 and the sensitive strain E. coli CCM 3953. The minimum inhibitory concentration was experimentally determined for these samples. Morphological changes upon exposure of the Fe16+AMP complex to E. coli CCM 3953 were also observed using a cryo-SEM scanning electron microscope. The development of resistance to the model bacterium E. coli TOP10 by transformation was also investigated and verified by polymerase chain reaction PCR and gel electrophoresis. Based on the results, it was proven that the Fe16+AMP complex has an inhibitory effect on the bacterium E. coli CCM 3953 and does not have a significant effect on the resistance spreading via transformation mechanism.
|
|
|
Preparation of nanoparticles and nanofibers with antimicrobial components
Kubišová, Veronika ; Slaninová, Eva (referee) ; Skoumalová, Petra (advisor)
This thesis addresses the problem of inadequate current wound therapy and presents a solution in the form of nanomaterial-based wound dressings (coverings). Specifically, it focuses on the development and characterization of various nanofibrous materials with integrated liposome particles that could serve as a source of therapeutic agents and be useful specifically in the field of a wound therapy. The review in the introductory part of the thesis first focused on the mentioned problem, which describes the shortcomings of existing conventional dressing materials. A description of human skin from an anatomical and functional point of view was not omitted, nor was the wound healing process itself. Different types of covering materials were also presented. However, a large part was focused on nanomaterials and their use in the field of the skin wound therapy. The nanomaterials mentioned were mainly liposome particles and nanofibres, as the experimental part of the work was focused on these structures. The description dealt with the characterization of these structures as well as the biopolymers used for their preparation. However, the aim was to prepare nanofibers with liposome content, so the method of forming such systems was described. Various therapeutic agents are also an integral part of the covering materials, especially those that suppress the development of infection and reduce wound pain; therefore, the search focused on the antibiotic ampicillin and the analgesic ibuprofen. The experimental part was devoted to the actual production of nanofibers with liposomes and also to the qualitative demonstration of the presence of liposomes in electrostatically prepared nanofibers. The selected polymeric components of these systems were polyhydroxybutyrate (PHB) and gelatine. However, the nanofibers and liposome particles (as well as combined liposomes with PHB) were first prepared in their own form and characterized mainly in terms of the gradual release of the drug substances. The results obtained were then compared with those of therapeutic drug delivery via combined nanofibrous structures with liposomes. For these combined structures, the aim was to achieve synergy in drug delivery between these systems. The aim of this work was to create a biomaterial covering with the controlled drug release. The drugs contained in these materials were the aforementioned ampicillin and ibuprofen. An important part of the work was then to determine the safety of the prepared materials which were tested for cytotoxicity, where the tests performed were MTT test and LDH test. And the actual wound healing ability of the nanofibers was then monitored in a scratch test or a "wound healing" test. At the end of the paper, recommendations for future work on this topic are given.
|
|
|
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.
|
|
| |
guest ::
