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Adhesion, growth and differentiation of skin cells on nanofibrous polymer membranes
Pajorová, Júlia ; Bačáková, Lucie (advisor) ; Eckhardt, Adam (referee)
Our study contributes to the tissue engineering, mainly to the construction of appropriate scaffolds for regeneration of damaged skin. Simultaneously, it brings valuable insights for basic research in the field of molecular mechanisms of adhesion, proliferation and phenotypic maturation of cells and the control of the cell behavior through the cell extracellular matrix (ECM), represented by synthetic nanofibrous material. Nanofibrous polylactic-co-glycolic acid (PLGA) membranes were prepared by needle-less electrospinning technology. These membranes were further modified with cell adhesion-mediating biomolecules, e.g. collagen, fibronectin and fibrin in order to increase their affinity to colonizing cells. Adhesion, growth and differentiation of keratinocytes (HaCaT) and fibroblasts, i.e. major cell types of epidermis and dermis, were evaluated on these nanofibrous membranes. The results show that the membrane modification using fibrin structures improved adhesion and proliferation of human dermal fibroblasts. The collagen structure on the surface of membranes improved the adhesion and proliferation of human HaCaT keratinocytes. Furthermore, fibrin structure stimulated fibroblasts to produce collagen, which is a major component of ECM in the natural skin dermis. Fibronectin enhanced cell attachment...
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Methodological study of changes in mechanical properties of collagen tissues from pes equinovarus congenitus
Červený, Gustav ; Lopot, František (advisor) ; Eckhardt, Adam (referee)
The goal of this thesis was to invent and perform a testing protocol, witch can detect connective tissue mechanical and structural properties simultaneously in patients with clubfoot. Based on literature findings, it was presumed that connective tissue differ between medial and lateral side of foot. Because of low availability of specimen, it was important to draw and mention crucial alterations of testing protokol, for minimization of failed measurments. For reasons above, two specimens diveded into two samples was used for experiment. Described and discused methodics may enable of reader to insigt to drawbacks of this examination. For usability of this protocol, a setting and testing of few hypotheses was performed. One axis tensial testing with SHG microscopy examination was used in combination for experiment. One of the main result was finding, that structural differencies witch were expected, were not distinctive in samples in untensioned state. But, distingtive differencies may be drawed in tensioned samples. This differences for low number of specimen, cannot show any tendence. Results of tensial testing showed, that samples from medial side of foot can have higher toughness and higher fragility. For future acquisition of tendencies in specimen differencies, it is important to set particular...
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Application of the stem cells in bone tissue engineering
Kročilová, Nikola ; Bačáková, Lucie (advisor) ; Eckhardt, Adam (referee)
Problems with musculoskeletal system, such as of developmental disorders, fractures or damage of the bone by age, inflammatory or tumor diseases, are still increasing in orthopaedics. Sometimes the bone tissue is not capable to completely regenerate to exert its physiological function in the organism. For this reason, using the bone replacements is necessary and common nowadays. Despite of an intensive research and testing of a wide range of the potential biomaterials and their combinations, the usage of metal materials for construction of the bone implants, still remains to be the gold standard. Ti-6Al-4V alloy is one of the commercialy used metal materials, which is known for the high mechanical and chemical resistance and a good biocompatibility. For a good biological response of the patient's organism for the bone implant, is an ability of osteointegration into the surrounding bone tissue, the key. This ability can be influenced in the case of the metals, by their surface structure. As it is known from earlier studies, the surface topography of the material is very important for the adhesion and proliferation of the bone cells, which are able to discriminate, very sensitively, between various stages of the material surface roughness. For this reason we have focused on studying of an influence...
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Cell growth on biomaterials for skin replacements and wound dressings
Kudláčková, Radmila ; Bačáková, Lucie (advisor) ; Rösel, Daniel (referee) ; Eckhardt, Adam (referee)
Tissue engineering is an emerging interdisciplinary field developing new ways of treatment of patient's tissue defects using artificial substitutes. Skin tissue engineering is developing skin substitutes and wound dressings that would replace current treatment using autologous, allogeneic or xenogenic substitutes. There are high demands on materials which should serve as a scaffolds for dermal fibroblasts and keratinocytes. They must be non-cytotoxic and biodegradable with a rate proportional to formation of a new tissue. The materials should support adhesion and proliferation of the cells and even they could release growth factors and antimicrobial substance to enhance healing and new tissue formation. In this master thesis, the cell adhesion and proliferation were evaluated on sodium carboxymethyl cellulose (Hcel® NaT), poly-ε-caprolactone (PCL), poly-L-lactide-co-ε-caprolactone (PLA/PCL) and cellulose acetate (AC) nanofiber membranes. Primary human dermal fibroblasts and HaCaT cell line keratinocytes were selected for evaluation. The cell adhesion was observed by fluorescent microscopy, the proliferation was determined by metabolic assay (WST-1) and the material cytotoxicity was evaluated in xCELLigence® system. Materials did not show cytotoxic effects on the cells. However, the materials did...
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