Výroba hydrogelových scaffoldů v kombinaci s vlákennými nosiči pro tkáňové inženýrství
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Date
2024-06-13
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Abstract
Jedním z klíčových faktorů tkáňového inženýrství (TI) je návrh biologicky aktivních scaffoldů s optimálními vlastnostmi pro danou tkáň. V poslední době se hydrogely těší značnému zájmu jako materiály pro přípravu scaffoldů pro TI díky své jedinečné kompoziční a strukturní podobnosti s přirozenou extracelulární matrix. Vyvinout scaffoldy pro regeneraci nervové tkáně je pro vědce velkou výzvou. Jedním z možných přístupů v tomto směru je využití právě hydrogelů. Ovšem díky velké komplexnosti nervové tkáně a velmi slabé přirozené regeneraci je nutné takovéto scaffoldy dále funkcionalizovat. V rámci této bakalářské práce je představena výroba hydrogelového scaffoldu na bázi želatiny síťovaného beta zářením v
kombinaci s orientovanými strukturami mikrovláken z polykaprolaktonu (PCL). K přípravě želatinového hydrogelu byla využita prasečí želatina s hodnotou Bloom 300, která byla síťována beta zářením dávkou 25 kGy, což je zároveň i sterilizační dávka v biomedicínské praxi. K výrobě mikrovláken byl využit PCL o molekulové hmotnosti 80 000 Da a zařízení na výrobu polymerních vláken tažením. Úspěšně se podařilo připravit kompozitní scaffold na bázi želatiny s mikrovlákny PCL na povrchu. Scaffold byl osazen buňkami. Experimenty ukázaly, že želatina síťovaná zářením beta je v porovnání s želatinou síťovanou glutaraldehydem biokompatibilnější. Fluorescenční mikroskopie dále ukázala, že buňky na materiálu proliferují. Dále byla patrná preference mikrovláken buňkami a orientace buněk podél orientovaných mikrovláken, nicméně došlo i k migraci po hydrogelu. Tyto výsledky naznačují, že vytvořený kompozitní scaffold by mohl sloužit jako základ pro materiál využívaný v tkáňovém inženýrství nervové tkáně.
One of the key factors in tissue engineering (TI) is the design of biologically active scaffolds with optimal properties for a given tissue. Recently, hydrogels have received considerable interest as materials for the preparation of scaffolds for TI due to their unique compositional and structural similarity to the natural extracellular matrix. Developing scaffolds for nerve tissue regeneration is a major challenge for scientists. One possible approach in this direction is the use of hydrogels. However, due to the high complexity of nerve tissue and the very poor natural regeneration, such scaffolds need to be further functionalized. This bachelor thesis presents the production of a gelatin-based hydrogel scaffold crosslinked by beta radiation combined with oriented microfibre structures made of polycaprolactone (PCL). Porcine gelatin with a Bloom value of 300 was used to prepare the gelatin hydrogel, which was cross-linked with beta radiation at a dose of 25 kGy, which is also the sterilization dose in biomedical practice. PCL with a molecular weight of 80,000 Da and equipment for the production of polymer fibers by drawing were used to produce microfibres. A gelatin-based composite scaffold \linebreak with PCL microfibers on the surface was successfully prepared. The Scaffold was seeded with cells. Experiments showed that radiation cross-linked beta gelatin is more biocompatible compared to glutaraldehyde cross-linked gelatin. Fluorescence microscopy further showed that cells proliferate on the material. Furthermore, the preference of the microfibers by the cells and the orientation of the cells along the oriented microfibers was evident; however, migration along the hydrogel also occurred. These results suggest that the composite scaffold created could serve as the basis for a material used in neural tissue engineering.
One of the key factors in tissue engineering (TI) is the design of biologically active scaffolds with optimal properties for a given tissue. Recently, hydrogels have received considerable interest as materials for the preparation of scaffolds for TI due to their unique compositional and structural similarity to the natural extracellular matrix. Developing scaffolds for nerve tissue regeneration is a major challenge for scientists. One possible approach in this direction is the use of hydrogels. However, due to the high complexity of nerve tissue and the very poor natural regeneration, such scaffolds need to be further functionalized. This bachelor thesis presents the production of a gelatin-based hydrogel scaffold crosslinked by beta radiation combined with oriented microfibre structures made of polycaprolactone (PCL). Porcine gelatin with a Bloom value of 300 was used to prepare the gelatin hydrogel, which was cross-linked with beta radiation at a dose of 25 kGy, which is also the sterilization dose in biomedical practice. PCL with a molecular weight of 80,000 Da and equipment for the production of polymer fibers by drawing were used to produce microfibres. A gelatin-based composite scaffold \linebreak with PCL microfibers on the surface was successfully prepared. The Scaffold was seeded with cells. Experiments showed that radiation cross-linked beta gelatin is more biocompatible compared to glutaraldehyde cross-linked gelatin. Fluorescence microscopy further showed that cells proliferate on the material. Furthermore, the preference of the microfibers by the cells and the orientation of the cells along the oriented microfibers was evident; however, migration along the hydrogel also occurred. These results suggest that the composite scaffold created could serve as the basis for a material used in neural tissue engineering.
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tkáňové inženýrství, hydrogely, drawing, želatina, polykaprolakton, síťování beta zářením