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Preparation and characterization of nanostructured resorbable substitutes for accelerated skin healing
Kacvinská, Katarína ; Muchová, Johana (oponent) ; Vojtová, Lucy (vedoucí práce)
Together with the increasing demands on the quality of treatment in the field of burn and plastic surgery, there is the possibility of applying new technological solutions in the treatment of defects with full loss of skin thickness. This thesis deals with a preparation of a nanostructured bilayer scaffold for skin tissue engineering, which substitutes a skin dermis (lower porous layer) and a basal membrane (upper nanofibrous thin layer)). The porous layer is based on collagen, which is also characterized in presence of different polysaccharide additives: chitosan, oxidized cellulose calcium salt (CaOC), carboxymethyl cellulose sodium salt (NaCMC)), as well as a dopamine and fibroblast growth factor (FGF) addition, in order to improve biomechanical properties, regulate and promote skin healing. Thin nanofibers layer consists of electrospun gelatin in combination with polycaprolactone (PCL) and CaOC. Two different fabrication mechanisms differing in cross-linking between the porous and nanofibrous layer are proposed. The scaffolds were evaluated in terms of biomechanical, structural and in-vitro properties. A uniaxial strain test has shown that the upper nanofibrous layer provides mechanical support, which is significantly enhanced with the polydopamine (PDA)-coated surface. Swelling test of porous layer showed adequate spaces to allow cells infiltration, what has been shown as decrease in presence of PDA. Degradation with collagenase and lysozyme has shown significant time prolongation and also proliferation and viability of the mouse fibroblast cells seeded on the scaffolds were significantly enhanced with PDA and FGF modification. Novel nanostructured bilayer scaffold possess good biomechanical properties and exhibis potential in skin tissue engineering by allowing cells to adhere, proliferate and generate expracellular matrix.
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Keramické materiály používané v medicíně - výzvy a příležitosti
Vitásek, Ladislav ; Sláma, Martin (oponent) ; Novotná, Lenka (vedoucí práce)
Tato bakalářská práce se zabývá keramickými materiály používanými v medicíně, zejména v ortopedických a dentálních aplikacích. Úvodní část práce se zaměřuje na požadavky a současný stav biomateriálů používaných v kostním tkáňovém inženýrství, jejich výhody a nevýhody. V další podkapitole jsou popsány kompozitní materiály, kde alespoň jednu složku tvoří keramický materiál. V samostatné části je popsána hutná keramika na bázi oxidu hlinitého a zhouževnatělého oxidu zirkoničitého. Hlavní část této práce je zaměřena na porézní keramické materiály na bázi kalciumfosfátů, jsou zde vymezeny požadavky na skafoldy, mezi které patří pórovitost, mechanické a biologické chování. Další část se zabývá způsoby přípravy keramických skafoldů. Experimentální část této práce byla věnována přípravě bioaktivního porézního keramického materiálu na bázi hydroxyapatitu a oxidu křemičitého se zlepšenými vlastnostmi. Replikační technikou byly vyrobeny 4 sady vzorků s obsahem oxidu křemičitého od 6 do 21 hm%. Připravené bioaktivní skafoldy se strukturou odpovídající spongiózní kosti se mohou uplatnit jako potenciální náhrady tvrdých tkání v kostním tkáňovém inženýrství.
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Bioceramic Materials for Advanced Medical Applications
Novotná, Lenka ; Lapčík,, Lubomír (oponent) ; Drdlíková, Katarina (oponent) ; Cihlář, Jaroslav (vedoucí práce)
The aim of this thesis was to prepare three-dimensional scaffolds that can be potentially used for the reconstruction and regeneration of damaged bone tissues. Two techniques were used to create the porous ceramic scaffolds – polymer replica technique and polymerization in situ. A variety of bioceramic materials were studied, mainly alumina, zirconia and calcium phosphates. The effect of processing parameters, such as composition of suspensions, as well as the effect of heat treatment on structure and final properties of the prepared scaffolds were evaluated. Morphology of the sintered scaffolds was characterised by scanning electron microscopy; in particular pore size, pores interconnection, total porosity and density were described. Phase composition, compressive strength and biological properties such as bioactivity and cytotoxicity were also discussed. The dissertation is divided into several sections. The literature review briefly summarizes the structure and properties of bones, the requirements for scaffolds, advantages and disadvantages of currently used materials and methods of ceramic foam preparation. The first part of experiments dealt with scaffolds prepared by polymer replica technique. All fabricated foams had interconnected pores with size in the range of 300 to 2000 m, total porosity was 50–99%. The compressive strength of calcium phosphate foams prepared by replica technique reached about 0.3 MPa (at 80 % porosity). Reinforcement of the scaffolds was reached by using bio-inert cores or by incorporation of silica into the composite structure. The strength of calcium phosphate/silica scaffold increased above 20 MPa. The last section of experimental part discusses in situ blown calcium phosphate scaffolds, created by using the formation process of polyurethane foam with diisocyanate and polyol components. The sintered scaffolds had mainly interconnected macroporous structure with pore size ranging 80 to 550 m. The total porosity was about 76 to 99 %. The advantage of this method compared to the polymer replica technique was that the struts were completely filled. None of the studied materials was cytotoxic and moreover all studied foams exhibited bioactive behaviour. The most promising adept for application in bone tissue engineering seems to be composite material containing calcium phosphates reinforced by silica.
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The effect of biologicaly active substances on the structure and properties of collagenous substrates
Muchová, Johana ; Michlovská, Lenka (oponent) ; Vojtová, Lucy (vedoucí práce)
The thesis deals with the preparation of 3D porous collagen scaffolds by freeze-drying and their modification with bioactive compounds. The natural polysaccharides, chitosan, calcium oxidized cellulose and chitin/chitosan-glucan complex for the modification have been used. The mechanical properties of the scaffolds have been enhanced by crosslinking process with carbodiimides. Growth factors have been delivered in the form of platelet lysate. The influence of biologically active additives, crosslinking agents, and enrichment with growth factors on the properties of the prepared scaffold and their bioactivity in tissues of living organisms have been investigated. Specifically, this study includes the morphological properties, structure, porosity, swelling stability, chemical composition, temperature of denaturation and biological properties. Scanning electron microscopy, infrared specktroscopy, differential scanning calorimetry and confocal microscopy have been used to the characterization. Prepared collagen substrates involving bioactive additive and platelet lysate could be used as scaffold for growing cells in systems with low mechanical loading and which has potential application in biomedicine.
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Effect of bioceramic additives on morphology, physical and biological properties of collagen scaffolds for bone tissue engineering
Klieštiková, Nikola ; Poláček, Petr (oponent) ; Brtníková, Jana (vedoucí práce)
The diploma thesis deals with preparation of three-dimensional porous collagen composite scaffolds for bone tissue engineering and study of the effect of addition of bioceramic particles on morphological, biomechanical and biological properties. Theoretical part describes biomaterials and bioceramic particles used for scaffolds in bone tissue engineering and their fabrications method. As for experimental part, samples were prepared by the freeze-drying method. As tested material, type I collagen from porcine and bovine sources was combined with hydroxyapatite and mixture of -tricalcium phosphate and -tricalcium phosphate in ratios 1 : 1, 1 : 2 and 2 : 1. The effect of bioceramics solubility and particle sizes on scaffolds morphology, biomechanics and biocompatibility was evaluated. Addition of bioceramic particles changed the morphology of the samples. The pore size decreased, whereas the porosity was nearly the same in all tested samples. Bioceramic particles also made the collagen matrix of the scaffolds less hydrophilic, moreover they stabilized the scaffolds against the effect of enzymatic degradation. The biomechanical properties of the samples were tested in both dry and hydrated state. In dry state, the pure bovine collagen scaffolds reached the highest compressive strength, contrary in hydrated state, the samples containing bioceramic particles reached the highest value. None of the samples was cytotoxic and the most preferable environment for cell adhesion and proliferation was in the pure bovine collagen scaffolds and also in the composite scaffolds with ratio HAp : -TCP : 1 : 1.
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Preparation and characterization of nanostructured resorbable substitutes for accelerated skin healing
Kacvinská, Katarína ; Muchová, Johana (oponent) ; Vojtová, Lucy (vedoucí práce)
Together with the increasing demands on the quality of treatment in the field of burn and plastic surgery, there is the possibility of applying new technological solutions in the treatment of defects with full loss of skin thickness. This thesis deals with a preparation of a nanostructured bilayer scaffold for skin tissue engineering, which substitutes a skin dermis (lower porous layer) and a basal membrane (upper nanofibrous thin layer)). The porous layer is based on collagen, which is also characterized in presence of different polysaccharide additives: chitosan, oxidized cellulose calcium salt (CaOC), carboxymethyl cellulose sodium salt (NaCMC)), as well as a dopamine and fibroblast growth factor (FGF) addition, in order to improve biomechanical properties, regulate and promote skin healing. Thin nanofibers layer consists of electrospun gelatin in combination with polycaprolactone (PCL) and CaOC. Two different fabrication mechanisms differing in cross-linking between the porous and nanofibrous layer are proposed. The scaffolds were evaluated in terms of biomechanical, structural and in-vitro properties. A uniaxial strain test has shown that the upper nanofibrous layer provides mechanical support, which is significantly enhanced with the polydopamine (PDA)-coated surface. Swelling test of porous layer showed adequate spaces to allow cells infiltration, what has been shown as decrease in presence of PDA. Degradation with collagenase and lysozyme has shown significant time prolongation and also proliferation and viability of the mouse fibroblast cells seeded on the scaffolds were significantly enhanced with PDA and FGF modification. Novel nanostructured bilayer scaffold possess good biomechanical properties and exhibis potential in skin tissue engineering by allowing cells to adhere, proliferate and generate expracellular matrix.
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Effect of bioceramic additives on morphology, physical and biological properties of collagen scaffolds for bone tissue engineering
Klieštiková, Nikola ; Poláček, Petr (oponent) ; Brtníková, Jana (vedoucí práce)
The diploma thesis deals with preparation of three-dimensional porous collagen composite scaffolds for bone tissue engineering and study of the effect of addition of bioceramic particles on morphological, biomechanical and biological properties. Theoretical part describes biomaterials and bioceramic particles used for scaffolds in bone tissue engineering and their fabrications method. As for experimental part, samples were prepared by the freeze-drying method. As tested material, type I collagen from porcine and bovine sources was combined with hydroxyapatite and mixture of -tricalcium phosphate and -tricalcium phosphate in ratios 1 : 1, 1 : 2 and 2 : 1. The effect of bioceramics solubility and particle sizes on scaffolds morphology, biomechanics and biocompatibility was evaluated. Addition of bioceramic particles changed the morphology of the samples. The pore size decreased, whereas the porosity was nearly the same in all tested samples. Bioceramic particles also made the collagen matrix of the scaffolds less hydrophilic, moreover they stabilized the scaffolds against the effect of enzymatic degradation. The biomechanical properties of the samples were tested in both dry and hydrated state. In dry state, the pure bovine collagen scaffolds reached the highest compressive strength, contrary in hydrated state, the samples containing bioceramic particles reached the highest value. None of the samples was cytotoxic and the most preferable environment for cell adhesion and proliferation was in the pure bovine collagen scaffolds and also in the composite scaffolds with ratio HAp : -TCP : 1 : 1.
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Modulace interakcí interleukinů a jejich receptorů
Nepokojová, Tereza ; Schneider, Bohdan (vedoucí práce) ; Obšilová, Veronika (oponent)
Skafoldy jsou proteiny s vysokou konformační stabilitou, díky kterým mohou být do určitých úseků proteinu zaváděny vícečetné mutace. I přes tyto mutace zůstává celková strukturální integrita proteinu zachována, stejně tak jeho fyzikálně-chemické vlastnosti. Mutacemi získává daný skafold nové vlastnosti, přičemž ve většině případů se jedná o vazebnou specifitu vůči předem určené molekule. Hlavní výhodou skafoldů je malá velikost, stabilita, nízká výrobní cena a snadnost přípravy. Skafold, se kterým bylo v této práci pracováno, je unikátní tím, že měl navržené dva vazebné povrchy, na kterých mohl být mutován. Každý z obou vazebných povrchů je možné nezávisle mutovat tak, aby na nich vzniklo vazebné místo pro jiný protein. V našem případě šlo o mutace vedoucí k vazbě dvou různých receptorů lidského cytokinu. Mutace se provádí pomocí kvasinkového displeje, jednou z metod řízené evoluce. Předmětem této diplomové práce je přenesení vazebných proteinů z kvasinkového expresního systému do systému bakteriálního, jejich produkce a purifikace a jejich následná charakterizace pomocí vybraných biofyzikálních metod. Pomocí těchto metod byla hodnocena stabilita struktury vazebných proteinů, jejich teplotní stabilita a vazebná afinita vůči oběma receptorům. Širším cílem projektu, jehož je tato práce součástí, je...
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The effect of biologicaly active substances on the structure and properties of collagenous substrates
Muchová, Johana ; Michlovská, Lenka (oponent) ; Vojtová, Lucy (vedoucí práce)
The thesis deals with the preparation of 3D porous collagen scaffolds by freeze-drying and their modification with bioactive compounds. The natural polysaccharides, chitosan, calcium oxidized cellulose and chitin/chitosan-glucan complex for the modification have been used. The mechanical properties of the scaffolds have been enhanced by crosslinking process with carbodiimides. Growth factors have been delivered in the form of platelet lysate. The influence of biologically active additives, crosslinking agents, and enrichment with growth factors on the properties of the prepared scaffold and their bioactivity in tissues of living organisms have been investigated. Specifically, this study includes the morphological properties, structure, porosity, swelling stability, chemical composition, temperature of denaturation and biological properties. Scanning electron microscopy, infrared specktroscopy, differential scanning calorimetry and confocal microscopy have been used to the characterization. Prepared collagen substrates involving bioactive additive and platelet lysate could be used as scaffold for growing cells in systems with low mechanical loading and which has potential application in biomedicine.
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Bioceramic Materials for Advanced Medical Applications
Novotná, Lenka ; Lapčík,, Lubomír (oponent) ; Drdlíková, Katarina (oponent) ; Cihlář, Jaroslav (vedoucí práce)
The aim of this thesis was to prepare three-dimensional scaffolds that can be potentially used for the reconstruction and regeneration of damaged bone tissues. Two techniques were used to create the porous ceramic scaffolds – polymer replica technique and polymerization in situ. A variety of bioceramic materials were studied, mainly alumina, zirconia and calcium phosphates. The effect of processing parameters, such as composition of suspensions, as well as the effect of heat treatment on structure and final properties of the prepared scaffolds were evaluated. Morphology of the sintered scaffolds was characterised by scanning electron microscopy; in particular pore size, pores interconnection, total porosity and density were described. Phase composition, compressive strength and biological properties such as bioactivity and cytotoxicity were also discussed. The dissertation is divided into several sections. The literature review briefly summarizes the structure and properties of bones, the requirements for scaffolds, advantages and disadvantages of currently used materials and methods of ceramic foam preparation. The first part of experiments dealt with scaffolds prepared by polymer replica technique. All fabricated foams had interconnected pores with size in the range of 300 to 2000 m, total porosity was 50–99%. The compressive strength of calcium phosphate foams prepared by replica technique reached about 0.3 MPa (at 80 % porosity). Reinforcement of the scaffolds was reached by using bio-inert cores or by incorporation of silica into the composite structure. The strength of calcium phosphate/silica scaffold increased above 20 MPa. The last section of experimental part discusses in situ blown calcium phosphate scaffolds, created by using the formation process of polyurethane foam with diisocyanate and polyol components. The sintered scaffolds had mainly interconnected macroporous structure with pore size ranging 80 to 550 m. The total porosity was about 76 to 99 %. The advantage of this method compared to the polymer replica technique was that the struts were completely filled. None of the studied materials was cytotoxic and moreover all studied foams exhibited bioactive behaviour. The most promising adept for application in bone tissue engineering seems to be composite material containing calcium phosphates reinforced by silica.
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