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The co-culture of keratinocytes and fibroblasts on a multi-layered polyester nanofibrous membrane enriched with platelet lysate
Blanquer, Andreu ; Filová, Elena ; Jenčová, V. ; Brož, Antonín ; Kuželová Košťáková, E. ; Lisnenko, M. ; Procházková, R. ; Bačáková, Lucie
The prevalence of chronic wounds is increasing due to the population ageing and specific illnesses like diabetes mellitus and vascular diseases. Nanofibrous membranes fabricated using synthetic polymers are promising materials to enhance skin wound healing. PCL and PVA membranes are being studied to be used as scaffolds for skin tissue engineering and hydrogels for controlled drug delivery, respectively. The present study considers the development of a multi-layered membrane made of PCL and PVA loaded with platelet lysate (PL). PCL nanofibers allowed cell adhesion and growth, whereas PVA acted as a hydrogel that releases the bioactive compounds of platelet lysate. The cytocompatibility of the membranes containing PL and without it was demonstrated on two cell types involved in wound healing, i.e. keratinocytes and fibroblasts. Both cell types were able to adhere and proliferate on the membranes. In addition, the membrane containing PL enhanced the proliferation of fibroblasts. A co-culture study was also performed by seeding each cell type on one side of the membrane. The cells were co-cultured for 7 days and the results showed that PL increased the proliferation of cells achieving a monolayer of keratinocytes or fibroblasts on each side of the membrane. Thus, the beneficial effect of PCL-PVA+PL membranes on monocultures and co-cultures of skin cells was demonstrated, and these membranes can be considered potential scaffolds for treatment of chronic wounds.
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Degradation of polycaprolactone electrospun materials - methods of analysis
Havlíčková, K. ; Řezanka, M. ; Kobera, Libor ; Kuželová Košťáková, E. ; Lukáš, D. ; Jenčová, V.
Biodegradable electrospun materials are widely used for medical application. Polycaprolatone is polymer suitable for electrospinning technology and is very often used to create nanofibrous scaffolds for tissue engineering. The time to disintegration or biodegradation of such materials is very important here. However, testing is not entirely easy. It is not possible to create exactly body-like conditions in vitro. Moreover, it is not easy to find suitable analytical methods that would show exactly what happens in the nanofibrous polycaprolactone electrospun samples at certain stages of degradation, ie how the internal structure of decaying nanofibers changes. This paper describes the traditional use of methods for testing polycaprolatone nanofibers by enzymatically catalysed degradation. Morphological changes are studied using scanning electron microscope images. However, it also offers a non-traditional analysis of polycaprolactone electrospun materials using the ssNMR method.
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In vitro testování buněčných nosičů na bázi nanovláken pro léčbu vitiliga
Kodedová, Barbora ; Amler, Evžen (vedoucí práce) ; Kuželová Košťáková, Eva (oponent)
Vitiligo je kožní onemocnění s prevalencí 2 % v celosvětové populaci. Je charakteristické sníženou aktivitou či ztrátou melanocytů v epidermis, vedoucí k depigmentaci v kůži a ve vlasech. Zásadně ovlivňuje psychiku, sociální vztahy pacientů a snižuje schopnost obrany organismu proti UV záření. Jednou z metod léčby je autologní transplantace melanocytů, nebo suspenze melanocytů s keratinocyty. Použití biokompatibilní membrány umožňující kultivaci těchto buněk s jejich následnou transplantací na depigmentované léze by mohlo zvýšit úspěšnost a zefektivnit tuto formu léčby. Cílem této práce proto bylo vytvořit biokompatibilní membránu z nanovlákenných vrstev polyvinylalkoholu (PVA), která by mohla sloužit jako nosič pro buněčné transplantáty v terapii vitiliga. Nosiče z PVA byly připraveny elektrostatickým zvlákňováním z hladiny a následně modifikovány studeným metanovým (CH4) plazmatem pro snížení jejich hydrofilnosti. Vzorky modifikovaných nanovlákenných nosičů byly podrobeny analýze fyzikálně-chemických vlastností (vizualizace morfologie vláken pomocí SEM, XPS analýza, smáčivost a Zeta potenciál povrchu). Následně byla sledována adheze, proliferace a metabolická aktivita myších linií melanocytů (Melan-a) a keratinocytů (XB2) pomocí testování in vitro. Z výsledků vyplývá, že krátkodobá modifikace CH4...
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