Studium vlivu přítomnosti proteinů na degradační chování polyesterových nanovlákenných materiálů
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Tkáňové inženýrství hraje důležitou roli v medicíně a zdravotní péči tím, že nabízí nové možnosti léčby a regenerace tkání. Studium scaffoldů v tkáňovém inženýrství je zásadní, protože scaffoldy jsou základní stavební jednotkou umožňující růst a diferenciaci buněk do specifických tkání. Zkoumání a optimalizace scaffoldů je pro dosažení úspěšných výsledků v oblasti tkáňového inženýrství klíčové. Jedním z důležitých aspektů u scaffoldů je jejich biodegradabilita a tato biodegradabilita může být ovlivněná adsorpcí proteinů na povrch materiálu ke které dochází ihned po implantaci materiálu do tkáně. V této práci byla studovaná enzymaticky katalyzovaná degradace materiálu PCL 80 za přítomnosti modelového proteinu BSA. Tato degradace byla posuzována pomocí gravimetrické metody hmotnostního úbytku a sledováním změny morfologie vláken na snímcích SEM. Dále byla pozorována i adsorpce proteinů na materiál v průběhu degradačního experimentu, a to spektrofotometricky a pomocí metod SDS-PAGE a FTIR. V rámci experimentální části práce byl zjištěn vhodný rozsah koncentrace enzymu v roztoku s proteiny pro pozorování enzymaticky katalyzované degradace za přítomnosti proteinů, tento rozsah byl stanoven na 10-20U/3ml. Bylo zjištěno, že při enzymaticky katalyzované degradaci materiálu za přítomnosti proteinů dochází ke dvěma souběžným procesům. K degradaci materiálu, která vede k restrukturalizaci povrchu materiálu a ke zvětšení jeho volného povrchu. A k adsorpci proteinů, která roste s volným povrchem, avšak adsorbované proteiny mají protektivní účinek a zabraňují přístupu enzymu k materiálu. Dále bylo v rámci experimentální části práce pozorováno, že na materiál se váží jak slabě, tak pevně vázané proteiny a některé pevně vázané proteiny zůstávají adsorbovány na materiál i po desorpci. Na materiál z roztoku enzymu s proteiny se též vážou i další frakce proteinů přítomné v roztoku BSA.
Tissue engineering plays an important role in medicine and healthcare by offering new possibilities for treatment and tissue regeneration. The study of scaffolds in tissue engineering is essential because scaffolds are the basic building blocks that enable the growth and differentiation of cells into specific tissues. Investigation and optimization of scaffolds is crucial to achieve successful results in tissue engineering. One of the important aspects of scaffolds is their biodegradability and this biodegradability can be affected by the adsorption of proteins onto the surface of the material which occurs immediately after implantation of the material into the tissue. In this work, the enzyme-catalyzed degradation of PCL 80 material in the presence of the model protein BSA was studied. This degradation was assessed by gravimetric mass loss method and by monitoring the change in fiber morphology on SEM images. Furthermore, the adsorption of proteins onto the material during the degradation experiment was also observed spectrophotometrically and by SDS-PAGE and FTIR. In the experimental part of the work, a suitable range of enzyme concentration in solution with proteins was determined for observing enzyme-catalyzed degradation in the presence of proteins, this range was set to 10-20U/3ml. It was found that two simultaneous processes occur in the enzyme catalyzed degradation of material in the presence of proteins. The degradation of the material leading to a restructuring of the material surface and an increase in the free surface area. And protein adsorption, which increases with the free surface, but the adsorbed proteins have a protective effect and prevent the enzyme from accessing the material. Further, it was observed in the experimental part of the work that both weakly and tightly bound proteins bind to the material and some tightly bound proteins remain adsorbed on the material even after desorption. Other fractions of proteins present in the BSA solution also bind to the material from the enzyme-protein solution.
Tissue engineering plays an important role in medicine and healthcare by offering new possibilities for treatment and tissue regeneration. The study of scaffolds in tissue engineering is essential because scaffolds are the basic building blocks that enable the growth and differentiation of cells into specific tissues. Investigation and optimization of scaffolds is crucial to achieve successful results in tissue engineering. One of the important aspects of scaffolds is their biodegradability and this biodegradability can be affected by the adsorption of proteins onto the surface of the material which occurs immediately after implantation of the material into the tissue. In this work, the enzyme-catalyzed degradation of PCL 80 material in the presence of the model protein BSA was studied. This degradation was assessed by gravimetric mass loss method and by monitoring the change in fiber morphology on SEM images. Furthermore, the adsorption of proteins onto the material during the degradation experiment was also observed spectrophotometrically and by SDS-PAGE and FTIR. In the experimental part of the work, a suitable range of enzyme concentration in solution with proteins was determined for observing enzyme-catalyzed degradation in the presence of proteins, this range was set to 10-20U/3ml. It was found that two simultaneous processes occur in the enzyme catalyzed degradation of material in the presence of proteins. The degradation of the material leading to a restructuring of the material surface and an increase in the free surface area. And protein adsorption, which increases with the free surface, but the adsorbed proteins have a protective effect and prevent the enzyme from accessing the material. Further, it was observed in the experimental part of the work that both weakly and tightly bound proteins bind to the material and some tightly bound proteins remain adsorbed on the material even after desorption. Other fractions of proteins present in the BSA solution also bind to the material from the enzyme-protein solution.
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tkáňové inženýrství, nanovlákenné materiály, PCL, biodegradace, proteinová adsorpce, lipáza