Abstract
This thesis describes micropatterning of Neuro-2a cell lines on modified chitosan substrates with microstructural pattern made through nanofabrication approaches to form neuronal networks. We performed methods of fabrication which combined with photolithography, inductively coupled plasma reactive ion etching (ICP-RIE), wet chemical etching and solvent casting to prepare six different types of chitosan substrates. Firstly, the neuronal cell-substrate interaction (flat and nanostructural) was investigated. We found that the projected cell area of Neuro-2a cells on flat chitosan substrates was larger than on nanostructural chitosan substrates and Neuro-2a cells preferred to adhere on flat chitosan surface region than on nanostructural chitosan substrates to immobilize and differentiation. Secondly, in order to grow neuronal network including somas patterning and neurites guidance, we have developed chitosan substrates, such as square pattern, single cell pattern, line pattern, negative chitosan substrates, network pattern and polarity induce pattern. We found that the polarity induce pattern was the most suitable approach to form neuronal network by controlling cell polarity in designed constrain geometry, and then, we designed grooves to fit the direction of neurites outgrowth. This research, we believe, would have the potential to study a wide range of neurobiological applications such as neurotransmitters screening and electrophysiological stimulation platforms. This development of chitosan-based platforms would contribute to biomedical engineering.