Příprava tkáňového nosiče pro náhradu nervové tkáně (míšní léze)
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V dnešní době je poranění páteře prakticky neléčitelné. Tato práce se zabývá výrobou náhrady, která by umožnila regeneraci míchy a pomohla obnovit její funkci. Implantát by měl mít podobu hydrogelového válce, ve kterém jsou podélně umístěna elektricky vodivá vlákna. Cílem této práce je vyrobit orientovaná vlákna opatřená vodivou povrchovou vrstvou a otestovat jejich cytokompatibilitu.Nejprve bylo provedeno in vitro testování PCL nanovláken opatřených vrstvami polypyrrolu různé tloušťky, jehož cílem bylo stanovit vhodnou vrstvu polypyrrolu. Poté byla vyrobena orientovaná PCL mikrovlákna metodou drawing. Vlákna byla potažena vrstvou polypyrrolu, zvolenou dle výsledků předchozího testování, a podrobena testování in vitro. Součástí in vitro testů byl test viability buněk, rastrovací elektronová mikroskopie a fluorescenční mikroskopie.Výsledky ukázaly, že vrstva polypyrrolu napomáhá adhezi a proliferaci buněk, příliš velká vrstva však inhibuje buněčný růst. Testování orientovaných vláken potvrdilo hypotézu, že buňky při růstu kopírují orientaci vláken. Tato práce je prvním krokem k výrobě nosiče pro regeneraci míchy. Výsledky potvrdily, že materiály vyrobené a testované v rámci této práce lze použít pro vývoj implantátu.
Nowadays a spinal cord injury cannot be cured. This work is focused on the development of the scaffold which enables regeneration of the spinal cord and helps to renew its function. Scaffold should have the form of a hydrogel cylinder with single conductive fibres inside. The aim of this work is to produce oriented fibres covered with a conductive layer and test its cytocompatibility.At first in vitro testing of electrospun PCL nanofibres with layers of polypyrrole with different thickness was done. Then were produced oriented PCL microfibers using the drawing method. Fibres were coated with a polypyrrole layer, which was elected on the basis of previous tests with electrospun material, and tested in vitro. The part of in vitro testing was cell viability test, scanning electron microscopy and fluorescence microscopy.The results showed that the polypyrrole coating supports the adhesion and cell proliferation, but thick layer inhibits the cell proliferation. Testing of oriented fibres confirms that cells copy the fibre orientation. This work is the first step in development of scaffold for spinal cord regeneration. Results confirmed that materials produced and tested in this work are useful for the implant.
Nowadays a spinal cord injury cannot be cured. This work is focused on the development of the scaffold which enables regeneration of the spinal cord and helps to renew its function. Scaffold should have the form of a hydrogel cylinder with single conductive fibres inside. The aim of this work is to produce oriented fibres covered with a conductive layer and test its cytocompatibility.At first in vitro testing of electrospun PCL nanofibres with layers of polypyrrole with different thickness was done. Then were produced oriented PCL microfibers using the drawing method. Fibres were coated with a polypyrrole layer, which was elected on the basis of previous tests with electrospun material, and tested in vitro. The part of in vitro testing was cell viability test, scanning electron microscopy and fluorescence microscopy.The results showed that the polypyrrole coating supports the adhesion and cell proliferation, but thick layer inhibits the cell proliferation. Testing of oriented fibres confirms that cells copy the fibre orientation. This work is the first step in development of scaffold for spinal cord regeneration. Results confirmed that materials produced and tested in this work are useful for the implant.
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míšní léze, drawing, polypyrrol, tkáňové inženýrství, scaffold, spinal cord injury, drawing, polypyrrole, tissue engineering, scaffold