微圖型化甲殼素基材用於神經元導引之發展

Development of Micropatterned Chitosan Substrates for Neuronal Guidance

宋淳諺(Chun-Yen Sung)

指導教授 ： 葉哲良 ； 鄭兆珉

國立清華大學/工學院/奈米工程與微系統研究所/碩士(2013年)

摘要

本篇論文設計出微圖型化表面之甲殼素基材排列神經母細胞株Neuro-2a，希望在基材上建立神經元連結網路。我們使用微奈米技術，如黃光微影、感應耦合式電漿反應離子蝕刻、濕式化學蝕刻和溶劑鑄造技術來製備六種具奈米結構圖案的甲殼素基材。
首先，我們探討神經母細胞株Neuro-2a和甲殼素基材 （平坦和全奈米結構）之間的相互作用關係。我們發現神經母細胞株Neuro-2a生長在平坦甲殼素基材其攤附面積大於在奈米結構甲殼素基材，且易於在平坦區攤附和生長。其次，我們為了在甲殼素基材上建立神經元連結網路，包含細胞本體排列和神經突觸導引，我們發展了六種型式的微圖型化甲殼素基材，包含矩形圖案、單一細胞圖案、線圖案、負片甲殼素基材、網狀圖案和極化圖案。研究中發現極化圖案是目前建立神經元連結網路最有效果之甲殼素基材。其原理先用表面形貌控制神經細胞極化方向，再設計溝槽方向符合神經突觸之分化方向，導引樹突和軸突連結。此研究將有潛力成為探討神經科學的平台，如神經傳導物質的測試和電生理刺激平台之應用，相信對生物醫學工程將是一大貢獻。

關鍵字

甲殼素基材 ； 黃光微影 ； 感應耦合式電漿反應離子蝕刻 ； 濕式化學蝕刻 ； 溶劑鑄造技術

並列摘要

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.

並列關鍵字

Chitosan substrates ； Neuro-2a cells ； ICP-RIE ； wet chemical etching ； solvent casting

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