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Encapsulation of active substances into nanofibers and possibilities of their application
Procházková, Lucie ; Pernicová, Iva (referee) ; Skoumalová, Petra (advisor)
The master thesis was based on the optimization of the production of nanofiber covers and to gaine the product for subsequent functional use. The production of nanofiber covers was made by electrospinning and forcespinning from selected materials. Polyhydroxybutyrate, gelatin, chitosan and alginate were used as starting materials. After successful optimization, these materials were enriched with active ingredients ampicillin and ibuprofen for the functionalized use of covers for more effective wound healing. The theoretical part was focused on the issue of skin, healing processes, types of wounds and nanofibers, the characterization of selected starting materials for the formation of nanofibers was also mentioned. The practical part was based on the lengthy optimization of the preparation of fiber covers and later enriched with active ingredients. Furthermore, combined covers made of different materials with contents of both active ingredients were designed. This was followed by the characterization of all prepared covers from the point of view of stability in the short and long term. The gradual release of active ingredients was determined spectrophotometrically and by hifh performance liquid chromatography. It was also important to determine the antimicrobial activity of selected active substances. At the end of all testing, combined coatings containing both active ingredients were used for safety testing with human keratinocyte cells (HaCaT). Safety testing was based on determining the viability of human cells using the MTT test, to verify the LDH test. A scratch test was also performed, a wound healing test after the application of devised combined covers.
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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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Porous hydrogels from natural gum modified with bio-active substances for accelerated wound healing
Černá, Eva ; Brtníková, Jana (referee) ; Vojtová, Lucy (advisor)
Hlavní téma této diplomové práce se zaměřuje na navržení, přípravu a optimalizaci složení hydrogelového krytu ran s vlastnostmi podobnými kůži. Finančně nenáročný hydrogelový kryt s vlastnostmi, který podporuje proces hojení, zabraňuje bakteriální infekci a snadná manipulace, je myšleným výsledkem této práce. Teoretická část je zaměřena na charakterizaci jednotlivých vrstev kůže, polymerní materiály vhodné pro léčbu ran, aditiva které podporují hojení a mají antimikrobiální vlastnosti, které tvoří aseptické prostředí v ráně. Experimentální práce zahrnuje několik kroků. Prvním z nich je optimalizace složení hydrogelového krytu ran složeného z přírodního polymeru, pryskyřice Gum Karaya (GK) modifikovaného se syntetickými hydrofilními gelujícími polymery a důležitou změkčující složkou pro získání měkkého, elastického a transparentního materiálu získaného metodou sušení, lyofilizace. Připravené filmy byly testování z hlediska botnání, hydrolitické stability, transparentnosti, pevnost v tahu v suchém i mokrém stavu, chemické složení a morfologie. Druhý krok zahrnoval integraci bioaktivního antiseptika do struktury hydrogelu, který zajišťuje vhodné antimikrobiální podmínky pro hojení ran. Připravené transparentní, hydrolyticky stabilní filmy se ukázaly být efektivní proti oběma testovaným druhům bakteriálních kmenů, gram-pozitivním i gram-negativním, mezi které patřily: Staphylococcus aureus methicillin-senzitivní, Staphylococcus aureus methicillin-rezistentní, Staphylococcus epidermidis, Klebsiella pneumoniae, Enterobacter cloacae, Enterococcus faecalis, Pseudomonas aeruginosa, Candida albicans and Escherichia coli. Následná práce bude zaměřena na testování in vitro a in vivo testy biocompatibility.
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Changing the role of a nurse in chronic wounds
LUSKOVÁ, Pavlína
The bachelor thesis is focused on the care of chronic wounds. It seeks to map the education of nurses in this field, to present the cooperation between physician and nurse, to introduce relationships in the workplace, to examine what influences physician and nurse in choosing material, such as success in wound healing, and last but not least to announce more products of wet healing. A chronic wound is a disorder of the integrity of the skin cover, which, despite adequate treatment for the period of 6-9 weeks, fails to show some signs of healing. Chronic wounds are a nursing problem and the care of a patient with a chronic wound carries certain specifics with it. Knowledge of phase healing and modern products are the main pillar of success, thanks to which it is possible to substantially help in the treatment of chronic wounds. The objective of the thesis was to underline the remarkable progress in the care of chronic wounds and also to get deeper into the whole issue. The bachelor thesis was divided into two parts, theoretical and empirical. The first part of this work introduces the concepts of chronic wound, the types of chronic wounds and the healing of wounds in general as well as the stages of healing and the means by which healing can be influenced. It also focuses on wound evaluation, wound dressing, document management, modern wet healing products, the role of nurses and, finally, education in this area. The second part of this work was dedicated to research investigation. Two goals were set in the practical part of the work. The first one: To map the cooperation between a physician and a nurse in the care of chronic wounds. The second one: To map the nurse's decision-making in the choice of material in the care of chronic wounds. Therein, four research questions have been identified: 1. How do nurse and physician work together in the care of chronic wounds? 2. What material do nurses most often use to heal chronic wounds? 3. What factors influence the nurse when choosing material in the care of chronic wounds? 4. What role does the nurse play in caring for a chronic wound? The research part was processed by a qualitative method using a semi-structured interview with selected nurses and physicians who take care of chronic wounds day-to-day. The interview consisted of 16-18 questions that were more elaborated during the interview so that respondents could present their ideas and impressions in relation to the topic more. The results obtained from the responses were then categorized and further processed. The results of the investigation showed that the cooperation between the nurse and the physician is generally effective, communication and trust between them is at a good level, and therefore the nurses are very strongly involved in the decision-making process regarding the choice of material. However, the cost of some products does not allow staff to work with them, although they find it particularly useful. Nevertheless, the wet healing method is economically less demanding, very effective and more comfortable for patients. It can also be stated that nurses are well trained with individual products and their workplaces offer them many opportunities in the development of education, which most of them are interested in. The conclusion of the work summarizes that a number of new products in the care of chronic wounds is still increasing, and therefore it is vital to constantly learn in this area. This work can serve as information material for people who are interested in this issue. It also brings with it useful opinions from nurses and physicians.
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Interactions of skin and stem cells with polymer nanofibres for construction of skin substitutes
Tomšů, Júlia ; Bačáková, Lucie (advisor) ; Sedmera, David (referee) ; Jendelová, Pavla (referee)
The skin is the largest organ of a human body with a crucial role in the maintenance of homeostasis; therefore any extensive skin injury leads to severe complications. Since the application of auto-, allo- and xeno-grafts is accompanied by severe problems like the source limitation and the graft rejection, a bioengineered skin substitute seems to be one of the promising healing approach. This work is focused mainly on the construction of a pre- vascularized skin substitute consisting of a collagen hydrogel reinforced by a biodegradable nanofibrous membrane. Another strategy described in this work is the development of temporary cellulose-based wound dressings. For both research strategies, various cell types were utilized, i.e. normal human dermal fibroblasts (NHDFs), human keratinocytes (hKs), adipose tissue-derived stem cells (ADSCs) and human umbilical vein endothelial cells (HUVECs). In order to enhance the cell adhesion and growth, the synthetic nanofibrous membranes were improved by protein nanocoatings. It was found out that NHDFs and ADSCs preferred fibrin nanocoatings, mainly thin fibrin homogeneous mesh on the surface of the membrane. Keratinocytes rather adhered and stratified on collagen substrates. These observations further motivated the construction of the bi-layered construct, where...
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Encapsulation of active substances into nanofibers and possibilities of their application
Procházková, Lucie ; Pernicová, Iva (referee) ; Skoumalová, Petra (advisor)
The master thesis was based on the optimization of the production of nanofiber covers and to gaine the product for subsequent functional use. The production of nanofiber covers was made by electrospinning and forcespinning from selected materials. Polyhydroxybutyrate, gelatin, chitosan and alginate were used as starting materials. After successful optimization, these materials were enriched with active ingredients ampicillin and ibuprofen for the functionalized use of covers for more effective wound healing. The theoretical part was focused on the issue of skin, healing processes, types of wounds and nanofibers, the characterization of selected starting materials for the formation of nanofibers was also mentioned. The practical part was based on the lengthy optimization of the preparation of fiber covers and later enriched with active ingredients. Furthermore, combined covers made of different materials with contents of both active ingredients were designed. This was followed by the characterization of all prepared covers from the point of view of stability in the short and long term. The gradual release of active ingredients was determined spectrophotometrically and by hifh performance liquid chromatography. It was also important to determine the antimicrobial activity of selected active substances. At the end of all testing, combined coatings containing both active ingredients were used for safety testing with human keratinocyte cells (HaCaT). Safety testing was based on determining the viability of human cells using the MTT test, to verify the LDH test. A scratch test was also performed, a wound healing test after the application of devised combined covers.
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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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Moist wound healing in a confrontation with the classical method of wound dressing
DVOŘÁKOVÁ, Vendula
The main focus of this thesis is the comparison of the classical wound healing method and the wet healing method. The latter method is being widely used nowadays but there are still departments healing wounds the classical way. And while the wet method is by far the more favourite one there are nurses who adamantly promote the classical one. The combination of both is also being used very often. The bachelor's thesis is divided in to two parts theoretical and empiric. First part is focusing on the anatomy of skin, wounds in general and the history of their categorization as well as the healing of wounds its phases and influencing factors. Furthermore, there are dressing options, evaluation and documentations of wounds, materials used for wet healing and education of nurses described. Second part contains the case study. Two goals were set in the empiric section. The first is to find a time difference in healing wounds using the classical and the wet method. The second one is to set a difference in the caretaking the methods demand. There were four research questions stated to support the goals. They are as follows: What issues are the nurses facing while using the classical wound healing method? What issues are the nurses facing while using the wet wound healing method? What kind of a dressing the nurses prefer? What is the time difference in healing while using these methods? Qualitative survey was used in processing the research part of the thesis. To collect data semi structured interviews were held with nurses dealing with wound dressing and chronical wound patient's case histories. There were 16 questions in the interview (Attachment 2) that were spontaneously extended by sub-questions. The resulting findings were categorized and further processed. The case studies focused on patients with chronic wounds, hospitalized in the aftercare unit and were extended by photo documentation. The results state that nurses prefer wet wound healing. This method was judged faster andmore comfortable for the patient as well as more efficient from the economical point of view. To be able to adequately work with the materials used for wet wound healing it is very important for the nurses to study the topic continuously. Further findings also show that the classical method or its elements are still being used to treat wounds and even there are positive aspects to be found. The nurses are still fond of certain classical wound healing materials therefore they keep coming back to this method. The research also shows that by combining both methods excellent results can be reached. This thesis is capable of functioning as informational material for those, who are interested in the topic of wound healing. Readers will find basic information about both methods of wound healing accompanied by opinions on this matter provided by professional nurses tasked with wound dressings. The case study part talks about the procedure of healing and the photos provided are suitable for comparing both wound healing methods.
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