Online-Marker der Gewebsentwicklung im kardiovaskulären Tissue Engineering : Evaluierung von sulfatierten Glykosaminoglykanen als Online-Marker der Extrazellularmatrix-Synthese im Kardiovaskulären Tissue Engineering
Tümen, Michael; Jockenhövel, Stefan (Thesis advisor)
Dissertation / PhD Thesis
The method of tissue engineering describes an interdisciplinary attempt of replacing injured or damaged tissue based on autologous cell constructs. This doctoral thesis explains the principals of tissue engineering and provides fundamental knowledge of important players of the Extracellular Matrix (ECM), their synthesis and their interactions. Furthermore, a non-destructive method to assess ECM development by detecting sulphated glycosaminoglycans (sGAGs) in the culture medium of statically cultivated cell-seeded fibrin gels is evaluated. The main results can be summarized as followed: - myofibroblasts isolated from human umbilical cord arteries are an appropriate cell source to investigate sGAG synthesis - the amount of sGAGs retained in the fibrin gels increases during the cultivation period - sGAGs cannot be detected in the culture medium with the here proposed assay, when cultivated statically without any mechanical stimulus - burst strength of fibrin gels decreases initially due to fibrinoylsis, but increases from the second quarter of cultivation period, when the synthesis of stabilizing ECM components like collagens or elastin rises - here proposed Blyscan® Assay is suitable for detection of sGAGs in cell-seeded fibrin gels, but has some serious disadvantages concerning the detection of small amounts of sGAGs and the missing possibility to distinguish between sGAG subtypes This doctoral thesis is part of the project „Online-Marker der Gewebsentwicklung im Kardiovaskulären Tissue Engineering“. To our knowledge, it is the first attempt to monitor ECM synthesis non-invasively by measuring biomarkers in the culture medium of cultivated fibrin gels.