Modifikace nanovlákenných tkáňových nosičů L-Argininem
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Cílem této práce byl vývoj, výroba a testování materiálů určených k hojení kožních poranění a použití pro kardiovaskulární systém. Pro testování byly vyrobeny nanovlákenné polykaprolaktonové vrstevy s aktivní složkou, viz grafický abstrakt. Aktivní látkou byl L-Arginin, který je přirozeným donorem oxidu dusnatého. Byly použity polykaprolaktonové vrstvy s koncentrací L-Argininu od 0 do10 hm%, byla charakterizována morfologie vrstev, byla provedena kvantifikace uvolňování L-Argininu z vrstev a biologické testování.L-Arginin měl vliv na morfologii nanovlákenné polykaprolaktonové vrstvy a zvyšoval plošnou hmotnost vrstev a hydrofobicitu. Oplach ethanolem a pufrem snižoval uvolněné množství aminokyseliny. Uvolňování aktivní látky ze sterilizovaných nanovlákenných vrstev bez oplachu probíhalo v pufru pozvolněji a výsledná koncentrace byla větší než u uvolňování v kompletním médium. Byl zkoumán vliv samotného L-Argininu i extraktů z materiálů modifikovaných L-Argininem na tři buněčné linie. Samotný L-Arginin byl pro fibroblasty a endotelové buňky toxický při koncentraci vyšší než 1 % a vyšší než 0,75 % pro hladskosvalové buňky. U nižších koncentracích L-Arginin nezvyšoval viabilitu buněk. Extrakty nanovlákenných vrstev byly odebírány v průběhu 14denního experimentu. Buňky byly inkubovány s extrakty, které byly odebrány v rozmezí 1 h až 14 dní. Viabilita buněk neklesla pod mez cytotoxicity kromě 14denního extraktu vrstvy s 10 hm% L-Argininu u fibroblastů.
The aim of the thesis was development, fabrication and testing materials designed for wound healing and cardiovascular applications. Nanofibrous polycaprolactone layers with an active component were produced, see the graphic abstract below. The active component was L-Arginine which is a natural donor of nitric oxide.Polycaprolactone layers with concentration of L-Arginine from 0 to 10 w% were used, its morphology was characterized, quantification of L-Arginine released from layers was assessed and biological testing was carried out.L-Arginine had an impact on morphology of the nanofibrous layer and increased its surface density and hydrophobicity. The amount of released amino acid was reduced after ethanol and buffer rinsing. The release of active component from sterilized nanofibrous layers without rinse processed in buffer gradually and the total concentration was higher than with release in complete media.The influence of L-Arginine dissolved in medium as well as extracts from electrospun materials was tested using three cell lines. L-Arginine itself was toxic for fibroblasts and endothelial cells in concentration higher than 1 % and higher than 0,75 % for smooth muscle cells. Lower concentration of L-Arginine did not increased viability of cells. Extracts of nanofibrous layers were stored for viability testing during 14 days experiment. Cells were incubated with materials extracts taken out in range of 1 hour to 14 days. Viability of cells decreased under cytotoxic 70 % only with fibroblasts incubated in 14 days extracts of layer containing 10 w% L-Arginine.
The aim of the thesis was development, fabrication and testing materials designed for wound healing and cardiovascular applications. Nanofibrous polycaprolactone layers with an active component were produced, see the graphic abstract below. The active component was L-Arginine which is a natural donor of nitric oxide.Polycaprolactone layers with concentration of L-Arginine from 0 to 10 w% were used, its morphology was characterized, quantification of L-Arginine released from layers was assessed and biological testing was carried out.L-Arginine had an impact on morphology of the nanofibrous layer and increased its surface density and hydrophobicity. The amount of released amino acid was reduced after ethanol and buffer rinsing. The release of active component from sterilized nanofibrous layers without rinse processed in buffer gradually and the total concentration was higher than with release in complete media.The influence of L-Arginine dissolved in medium as well as extracts from electrospun materials was tested using three cell lines. L-Arginine itself was toxic for fibroblasts and endothelial cells in concentration higher than 1 % and higher than 0,75 % for smooth muscle cells. Lower concentration of L-Arginine did not increased viability of cells. Extracts of nanofibrous layers were stored for viability testing during 14 days experiment. Cells were incubated with materials extracts taken out in range of 1 hour to 14 days. Viability of cells decreased under cytotoxic 70 % only with fibroblasts incubated in 14 days extracts of layer containing 10 w% L-Arginine.
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L-Arginin, Oxid dusnatý, Polykaprolakton, Elektrostatické zvlákňování, Cílené dodávání léčiv, L-Arginine, Nitric oxide, Polycaprolactone, Electrospinning, Drug delivery system