Příprava a charakteristika vlákenného nosiče pro náhradu nervové tkáně
Title Alternative:Preparation and characterization of the scaffold for the nerve tissue engineering
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V dnešní době jsou stále nemoci a zranění, která medicína zatím nedokáže vyléčit, včetně poranění míchy. Tato práce se zabývá výrobou a charakteristikou nosiče, který by umožnil regeneraci míchy a pomohl tak obnovit její funkci. Konečný implantát by měl mít podobu hydrogelového válce, ve kterém jsou podélně inkorporovaná elektricky vodivá vlákna. Cílem práce je vyrobit jednotlivá orientovaná vlákna metodou drawing modifikovaná vodivou povrchovou vrstvou, popsat vlastnosti materiálu a otestovat jeho cytokompatibilitu.Metodou drawing byla vyrobena jednotlivá orientovaná PCL vlákna a opatřena vrstvou polypyrrolu. Při charakterizaci materiálu byla měřena viskozita a povrchové napětí roztoku, z něhož byla vlákna vyrobena. Dále byla zkoumána morfologie PCL vláken, jejich pevnost v tahu a degradační chování, na vláknech s povrchovou vrstvou polypyrrolu byla měřena vodivost. Následně byla vlákna podrobena in vitro experimentům, jejichž součástí byl test metabolické aktivity buněk a fluorescenční mikroskopie, kterou bylo možné sledovat i morfologii buněk. Pro in vitro testy byly jako kontrolní vzorky použity také neorientované vrstvy vláken vyrobené metodou elektrospinning a jednotlivá orientovaná vlákna vyrobená z kopolymeru PLCL.Výsledky ukázaly, že buňky na orientovaných vláknech modifikovaných povrchovou vrstvou polypyrrolu dobře proliferují a kopírují orientaci vláken. Tento vlákenný materiál by proto měl být dobrou oporou pro orientovaný růst nervových buněk a tedy vhodnou součástí nosiče pro regeneraci nervové tkáně. Jednotlivá orientovaná mikrovlákna mohou být klíčovým materiálem pro regeneraci míchy i nervové tkáně obecně.
Nowadays, there are still diseases and injuries that medicine cannot cure, including spinal cord injuries. This work is focused on the production and characterization of the scaffold, which would allow regeneration of the spinal cord and help to renew its function. Scaffold should have a form of a hydrogel cylinder with incorporated oriented conductive fibres. The aim of the work is to produce by the drawing method individual oriented fibres, modify them by the conductive surface layer, describe the properties of the material and test the cytocompatibility of the fibres. Individual oriented PCL fibres were produced by the drawing method and they were modified with a polypyrrole layer. When characterizing the material, the viscosity and surface tension of the polymer solution were measured. In addition, the morphology of the fibres, their tensile strength and degradation behaviour were studied, on the polypyrrole-coated fibres the conductivity was measured. The fibres were subjected to in vitro experiments, including cell viability test and fluorescent microscopy, which shows the cell morphology. For in vitro tests were used also fibres with random orientation and individual PLCL oriented fibres. The results showed that cells on polypyrrole-coated oriented fibres proliferate well and they copy the orientation of the fibres. The fibrous material should be a good support for orientated nerve cell growth and therefore a suitable part of the scaffold for nerve tissue regeneration. Individual oriented microfibres can be a key material for nerve tissue regeneration and cure for the spinal cord injury.
Nowadays, there are still diseases and injuries that medicine cannot cure, including spinal cord injuries. This work is focused on the production and characterization of the scaffold, which would allow regeneration of the spinal cord and help to renew its function. Scaffold should have a form of a hydrogel cylinder with incorporated oriented conductive fibres. The aim of the work is to produce by the drawing method individual oriented fibres, modify them by the conductive surface layer, describe the properties of the material and test the cytocompatibility of the fibres. Individual oriented PCL fibres were produced by the drawing method and they were modified with a polypyrrole layer. When characterizing the material, the viscosity and surface tension of the polymer solution were measured. In addition, the morphology of the fibres, their tensile strength and degradation behaviour were studied, on the polypyrrole-coated fibres the conductivity was measured. The fibres were subjected to in vitro experiments, including cell viability test and fluorescent microscopy, which shows the cell morphology. For in vitro tests were used also fibres with random orientation and individual PLCL oriented fibres. The results showed that cells on polypyrrole-coated oriented fibres proliferate well and they copy the orientation of the fibres. The fibrous material should be a good support for orientated nerve cell growth and therefore a suitable part of the scaffold for nerve tissue regeneration. Individual oriented microfibres can be a key material for nerve tissue regeneration and cure for the spinal cord injury.