Medizinische Universität Graz Austria/Österreich - Forschungsportal - Medical University of Graz
Gewählte Publikation:
Grossmann, T.
Development and application of in vitro cell culture models to investigate vocal fold physiology and pathophysiology on the molecular level.
Doktoratsstudium der Medizinischen Wissenschaft; Humanmedizin; [ Dissertation ] Graz Medical University; 2020. pp. 153
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- Autor*innen der Med Uni Graz:
- Grossmann Tanja
- Betreuer*innen:
- Gugatschka Markus
- Kamolz Lars-Peter
- Obermayer-Pietsch Barbara
- Altmetrics:
- Abstract:
- Cell culture is a valuable, indispensable tool for controlled biomedical research, enabling a better insight into the cellular and molecular mechanisms underlying vocal fold function in health, disease and injury. To date, most of the current in vitro applications has been limited by the use of a single cell type: fibroblasts. This main cellular component of the vocal fold connective tissue, the lamina propria, has been described to produce large amounts of the extracellular matrix, thereby substantially maintaining the unique tissue characteristics. Regarding the complex multilayered and multicellular microstructure of native vocal folds, its mechanical versatility during sound production and biophysical microenvironment, enhancement of traditional two-dimensional and existing three-dimensional in vitro model systems has to be established. The aim of this thesis was to design possible strategies to develop and apply enhanced cell culture models in vitro. Therefore, we established an effective isolation procedure of near primary epithelial cells out of human oral mucosa and used these cells to bioengineer three-dimensional constructs, together with vocal fold fibroblasts, under organotypic conditions. We could show that key morphologic characteristics of co-cultivated constructs resembled the native mucosa, including a mulitlayered epithelium and a continuous basement membrane. Furthermore, we utilized a recently engineered phonomimetic bioreactor to explore the isolated or combined effects of vibration and cigarette smoke on the cellular level of immortalized human vocal fold fibroblasts. Altogether, the utilized and established strategies will provide innovative, stable and reproducible in vitro models. They will substantially contribute to a deeper understanding of vocal fold micro-(patho)physiology, provide an in vitro platform for preclinical drug testing and may serve as a graft for laryngeal mucosal transplantation.