Date

2022-8-29

Author

Culfa, Şeyda

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Cells' interaction with their environment and how their responses to different mechanical cues are regulated have attracted attention for decades. These mechanical cues vary from topography to substrate stiffness. Knowing how topography-induced cell responses are regulated might give useful information about cell behaviors in health and disease states. This knowledge can be utilized to manipulate cell behaviors for different purposes. Topography-dependent cell behaviors rely on various parameters such as cell type, substrate material type, and topographical features. The Neuro-2A (N2a) cell line was used in the present study because of its availability and versatility. Two different silicon substrate types, polished or laserfabricated, were used. Fabrication of Silicon was carried out by ultrafast infrared pulsed laser to create micro- and/or nano-topographies. Scanning Electron Microscopy (SEM) and Atomic Force Microscopy were used to characterize substrates. Cells were grown on a control substrate (plastic dish - 35 mm x 10 mm TC- treated plastic cell culture dish which is made up of polystyrene), polished silicon, and laser-fabricated substrates with different topographies for 24 hours to evaluate cell behaviors. After 24 hours, cell morphological responses were studied with confocal microscopy and SEM. Also, after 24 hours, focal-adhesion-related gene expression levels were measured with qRT-PCR and protein expression levels with Western Blot analysis. On all topographies, there was a significant increase in the majority of focal adhesion-related gene expression levels compared to control cells grown on the plastic dish. It was also observed that actin fibers were aligned with the direction of the lines of topographies, especially when two different topographies were combined on a single substrate. When the cells are differentiated, this directional alignment of actin fibers becomes more prominent.In conclusion, substrate topography did not affect only the cells' morphological behaviors but also had a critical role in the changes at the gene and protein levels. Also, it was found that substrate topography has different effects on different cell behaviors associated with spreading and adhesion. Therefore, manipulating these behaviors at the gene and protein levels might be a promising approach for studying cell behaviors in health and disease states in future studies.

Subject Keywords

Neuro-2A cells, focal adhesion-related genes, surface topography, silicon, imaging, mechanotransduction

URI

https://hdl.handle.net/11511/98757

Collections

Graduate School of Natural and Applied Sciences, Thesis