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Interaction of bone and vascular cells with materials constructed for tissue replacement and drug delivery system
Vandrovcová, Marta ; Bačáková, Lucie (advisor) ; Smetana, Karel (referee) ; Jirka, Ivan (referee)
Interactions between biomaterials and cells play an important role in tissue engineering. It has been repeatedly shown that the cell behaviour strongly depends on physical and chemical properties of the material surface. In our study we focused on materials used for the construction of bone implants and replacements to support cell adhesion, growth and osteogenic diferenciation and to lead to an integration between an implant and a bone tissue. First we tested an influence of different micrpattern of fullerens' C60 and composites' C60/Ti films to adhesion of bone cells MG 63, their initial spreading, growth, viability and formation of cytosceletal protein actin. Some of these films were additionally iradiated with Au+ ions, which led to the conversion of some C60 molcules into amorphous carbon (a-C). We confirmed that pattern influenced the distribution of cells without decrease viability. Thus, the production of patterns could be used to direct cell adhesion. In the second study, we also observed the influence of the thickness of coated layer. We found that more selective cell growth can be achieved by depositing of thicker fullerens' film. Our study with osteoblast-like MG 63 cells cultured on PLGA films revealed, that surface coated with chosen components of extracellular matrix (colagen I and...
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Studium interakce buněk s biomimetickým materiálem a jeho využití v biomedicíně
Sauerová, Pavla ; Hubálek Kalbáčová, Marie (advisor) ; Vandrovcová, Marta (referee) ; Jendelová, Pavla (referee)
Biomaterials are considered as very promising tools for regenerative medicine. They have compensatory or supporting function in organism and they are often developed to support specific conventional medical procedures. So-called biomimetic materials are developed to imitate natural environment of organism and to induce positive innate responses of organism. An essential part of biomaterial development is in vitro biological evaluation, which characterizes (often for the first time) the potential of developed material for its clinical application. This Ph.D. thesis deals with in vitro biological evaluation of three different biomimetic materials. In all three cases, the comprehensive evaluation was an integral part of the material development and optimization processes. Each material was in vitro characterised at the level of cell-material interactions with respect to its intended specific application.. In the first part, cell response to potential drug delivery system based on colloidal complexes of cationic surfactants with hyaluronic acid (HyA) was characterized. HyA protection ability and its limits were described; also the role of fetal bovine serum (FBS) in cell response to the stress stimuli was confirmed. Results considered surfactant-HyA complexes as promising system for drug delivery. In...
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Interaction of bone and vascular cells with materials constructed for tissue replacement and drug delivery system
Vandrovcová, Marta
Interactions between biomaterials and cells play an important role in tissue engineering. It has been repeatedly shown that the cell behaviour strongly depends on physical and chemical properties of the material surface. In our study we focused on materials used for the construction of bone implants and replacements to support cell adhesion, growth and osteogenic diferenciation and to lead to an integration between an implant and a bone tissue. First we tested an influence of different micrpattern of fullerens' C60 and composites' C60/Ti films to adhesion of bone cells MG 63, their initial spreading, growth, viability and formation of cytosceletal protein actin. Some of these films were additionally iradiated with Au+ ions, which led to the conversion of some C60 molcules into amorphous carbon (a-C). We confirmed that pattern influenced the distribution of cells without decrease viability. Thus, the production of patterns could be used to direct cell adhesion. In the second study, we also observed the influence of the thickness of coated layer. We found that more selective cell growth can be achieved by depositing of thicker fullerens' film. Our study with osteoblast-like MG 63 cells cultured on PLGA films revealed, that surface coated with chosen components of extracellular matrix (colagen I and...
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Development of extracellular-matrix scaffolds for CNS repair
Výborný, Karel ; Kubinová, Šárka (advisor) ; Hampl, Aleš (referee) ; Vandrovcová, Marta (referee)
Výborný, Karel. Vývoj materiálů na bázi extracelulární matrix pro léčbu centrálního nervového systému. [Development of extracellular-matrix scaffolds for CNS repair]. Praha, 2020. 105 stran, 3 přílohy. Dizertační práce (Ph.D.). Univerzita Karlova, 2. lékařská fakulta, Ústav experimentální medicíny, AV ČR v.v.i. Abstract Brain and spinal cord injury are a serious traumata and despite intensive research, there is still no effective treatment for patients. One of the studied approaches is to use various biomaterials to repair the damaged neural tissue. The aim of this thesis is to study the regenerative and neurotrophic effects of injectable extracellular matrix (ECM) hydrogels prepared by decellularization from porcine tissue (brain, spinal cord, bladder) and human umbilical cord (UC) in reconstruction of damaged neural tissue of the brain and spinal cord in rats. We characterized ECM hydrogels in terms of mechanical and rheological properties, structure and composition, cell adhesion, migration and proliferation. In a model of spinal cord hemisection, we compared the regenerative effect of ECM hydrogels derived from porcine spinal cord and urinary bladder. We found no benefits of tissue specific ECM prepared from the tissue of neural origin in terms of its neurotrophic properties in vitro or in vivo....
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Studium interakce buněk s biomimetickým materiálem a jeho využití v biomedicíně
Sauerová, Pavla ; Hubálek Kalbáčová, Marie (advisor) ; Vandrovcová, Marta (referee) ; Jendelová, Pavla (referee)
Biomaterials are considered as very promising tools for regenerative medicine. They have compensatory or supporting function in organism and they are often developed to support specific conventional medical procedures. So-called biomimetic materials are developed to imitate natural environment of organism and to induce positive innate responses of organism. An essential part of biomaterial development is in vitro biological evaluation, which characterizes (often for the first time) the potential of developed material for its clinical application. This Ph.D. thesis deals with in vitro biological evaluation of three different biomimetic materials. In all three cases, the comprehensive evaluation was an integral part of the material development and optimization processes. Each material was in vitro characterised at the level of cell-material interactions with respect to its intended specific application.. In the first part, cell response to potential drug delivery system based on colloidal complexes of cationic surfactants with hyaluronic acid (HyA) was characterized. HyA protection ability and its limits were described; also the role of fetal bovine serum (FBS) in cell response to the stress stimuli was confirmed. Results considered surfactant-HyA complexes as promising system for drug delivery. In...
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Growth factors and other bioactive substances for osteogenic differentiation of mesenchymal stem cells
Blahnová, Veronika ; Rampichová, Michala (advisor) ; Vandrovcová, Marta (referee)
The main function of mesenchymal stem cells in the body is to facilitate the restoration and regeneration of damaged tissues. They are known for the ability to differentiate into tissue originating from the mesoderm, which among others includes connective tissues. Due to this feature are MSCs being intensively examined. Different directions of differentiation can be induced by treatment of specific polypeptides, so called growth factors. In the field of tissue engineering are growth factors used to induce and accelerate the healing processes. They may be incorporated into the nanofiber carrier which is inserted into the site of injury. Cells in this area would thus be stimulated by surrounding 3D microenvironment. At the same time the scaffold provides a supply of growth factors which are able to affect metabolism, motility and differentiation of present cells. In order to induce osteogenic differentiation of human MSCs the following bioactive substances were used: TGF-β, bFGF, HGF, IGF-1, VEGF and the BMP-2 and the organic acid taurine. During 21 days lasting experiments, were these molecules added to the medium in various combinations and in the case of taurine also at two different concentrations. Cells were cultured on plastic. The best effect on cellular metabolism of MSCs, evaluated by MTS...
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Interaction of bone and vascular cells with materials constructed for tissue replacement and drug delivery system
Vandrovcová, Marta
Interactions between biomaterials and cells play an important role in tissue engineering. It has been repeatedly shown that the cell behaviour strongly depends on physical and chemical properties of the material surface. In our study we focused on materials used for the construction of bone implants and replacements to support cell adhesion, growth and osteogenic diferenciation and to lead to an integration between an implant and a bone tissue. First we tested an influence of different micrpattern of fullerens' C60 and composites' C60/Ti films to adhesion of bone cells MG 63, their initial spreading, growth, viability and formation of cytosceletal protein actin. Some of these films were additionally iradiated with Au+ ions, which led to the conversion of some C60 molcules into amorphous carbon (a-C). We confirmed that pattern influenced the distribution of cells without decrease viability. Thus, the production of patterns could be used to direct cell adhesion. In the second study, we also observed the influence of the thickness of coated layer. We found that more selective cell growth can be achieved by depositing of thicker fullerens' film. Our study with osteoblast-like MG 63 cells cultured on PLGA films revealed, that surface coated with chosen components of extracellular matrix (colagen I and...
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Interaction of bone and vascular cells with materials constructed for tissue replacement and drug delivery system
Vandrovcová, Marta ; Bačáková, Lucie (advisor) ; Smetana, Karel (referee) ; Jirka, Ivan (referee)
Interactions between biomaterials and cells play an important role in tissue engineering. It has been repeatedly shown that the cell behaviour strongly depends on physical and chemical properties of the material surface. In our study we focused on materials used for the construction of bone implants and replacements to support cell adhesion, growth and osteogenic diferenciation and to lead to an integration between an implant and a bone tissue. First we tested an influence of different micrpattern of fullerens' C60 and composites' C60/Ti films to adhesion of bone cells MG 63, their initial spreading, growth, viability and formation of cytosceletal protein actin. Some of these films were additionally iradiated with Au+ ions, which led to the conversion of some C60 molcules into amorphous carbon (a-C). We confirmed that pattern influenced the distribution of cells without decrease viability. Thus, the production of patterns could be used to direct cell adhesion. In the second study, we also observed the influence of the thickness of coated layer. We found that more selective cell growth can be achieved by depositing of thicker fullerens' film. Our study with osteoblast-like MG 63 cells cultured on PLGA films revealed, that surface coated with chosen components of extracellular matrix (colagen I and...
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Growth and differentiation of human osteoblast-like cells on TI-6AL-4V alloy modified with anodization
Doubková, Martina ; Bačáková, Lucie ; Pařízek, Martin ; Vandrovcová, Marta ; Musílková, Jana ; Lisá, Věra ; Gabor, R. ; Marvan, J.
Titanium and its alloys are widely used as hard tissue implants for their high biocompatibility and suitable physical properties. Along with other surface treatment methods, anodization technique is known to enhance osseointegration. The aim of this study is to evaluate the adhesion, growth and osteogenic differentiation of human osteoblast-like SAOS-2 cells on Ti-6Al-4V samples anodized in electrolytes composed of a stable volume of KOH (336.48 g/l) and variable volumes of liquid glass (124.47 g/l, 84,12 g/l and 37.38 g/l; samples s5, s6 and s7, respectively). Non-anodized Ti-6Al-4V samples, cell culture polystyrene (PS) and microscopic glass coverslips served as control materials. On days 2 and 4 after seeding, the cell number did not differ significantly among the tested samples. However, on day 7, the cell number on s6 samples reached the lowest values, which could be attributed to a non-homogeneous TiO2 film on s6 samples formed during anodization. Nevertheless, the osteogenic differentiation, estimated by the intensity of fluorescence of collagen I in cells grown in a differentiation medium, was the highest on s6 samples. On s5 samples, coated with homogeneous TiO2 films, both cell numbers and intensity of fluorescence of collagen I was relatively high. The bone matrix mineralization, evaluated by Alizarin Red staining, was the highest on s5 samples in standard culture medium, and similar on all tested samples in differentiation medium. Thus, the surface modification of s5 samples could be considered the most suitable for application in bone implants.
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