Studium adsorpce proteinů na polyesterové mikro a nanovlákenné materiály
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Význam proteinové adsorpce při tvorbě extracelulární hmoty, jejím osazení buňkami a následné obnově tkání je zcela zásadní. Zatímco v oblasti nanočástic je tento jev již dobře prozkoumán, v oblasti vláken-ných scaffoldů je tato disciplína stále prozkoumána nedostatečně. Různé vlastnosti specifické vlákenným materiálům ovlivňují průběh proteinové adsorpce a nepřímo tak určují reakci organismu. V této diplomové práci byla testována adsorpce albuminu z hovězího séra na několika polyesterových nano- a mikro-vlákenných materiálech. Byl hledán vhodný postup adsorpce a desorpce, ale také nejvhodnější způsob následné analýzy adsorbovaných proteinů. Několik metod analýzy bylo zkoumáno jak z hlediska náročnosti, tak z hlediska správnosti výsledků. Na základě výsledků byly mezi sebou porovnány jak materiály, tak analytické metody. Kromě standardních metod elektroforézy a chromatografie se ukázala pro větší množství vzorků být vhodnou metoda fluorimetrická. Z testovaných materiálů se vhodným k dalšímu zkoumání jevil mikrovlákenný PCL 80 pro jeho vysoké hodnoty adsorbovaného albuminu, avšak hydrofobní blend s PLCL v poměru 1:1 váže albumin silněji.
The protein adsorption is a crucial process at the formation of extracellular matrix, its cell adhesion and proliferation and resulting tissue renewal. In connection with nanoparticles, this phenomenon is well understood, whereas in the field of fibrous scaffolds, this phenomenon has yet to be investigated. Diverse properties of fibrous materials affect the process of protein adsorption, indirectly determining the response of an organism. In this diploma thesis, adsorption of bovine serum albumin onto several polyester nano- and micro-fibrous materials has been examined. A process of adsorption and desorption was determined and also the most eligible analysis methods of adsorbed proteins were sought. Several methods of analysis have been investigated in terms of method feasibility and results accuracy. Based on the results, both materials were compared between each other as well as the analytical methods used. In addition to the standard methods of electrophoresis and chromatography, the fluorimetry method proved to be suitable for analysis of large number of samples. From materials available, the most eligible for further research would be micro-fibrous PCL 80 due to high amounts of adsorbed proteins, but hydrophobic blend 1:1 with PLCL bonds with albumin stronger.
The protein adsorption is a crucial process at the formation of extracellular matrix, its cell adhesion and proliferation and resulting tissue renewal. In connection with nanoparticles, this phenomenon is well understood, whereas in the field of fibrous scaffolds, this phenomenon has yet to be investigated. Diverse properties of fibrous materials affect the process of protein adsorption, indirectly determining the response of an organism. In this diploma thesis, adsorption of bovine serum albumin onto several polyester nano- and micro-fibrous materials has been examined. A process of adsorption and desorption was determined and also the most eligible analysis methods of adsorbed proteins were sought. Several methods of analysis have been investigated in terms of method feasibility and results accuracy. Based on the results, both materials were compared between each other as well as the analytical methods used. In addition to the standard methods of electrophoresis and chromatography, the fluorimetry method proved to be suitable for analysis of large number of samples. From materials available, the most eligible for further research would be micro-fibrous PCL 80 due to high amounts of adsorbed proteins, but hydrophobic blend 1:1 with PLCL bonds with albumin stronger.
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proteinová adsorpce, vlákenné scaffoldy, albumin, polyestery, proteinová korona, protein adsorption, fibrous scaffolds, albumin, polyesters, protein corona