本論文係以奈米3D微影技術（Nano 3D Lithography, N3L）製做生物支架，以往以雙光子聚合技術（Two-photo Polymerization, TPP）製作之生物支架因聚焦光點小，導致需要加工時間過長，使生物支架尺寸會被限制在50微米左右，而奈米3D微影技術透過劑量累積效應，可縮短加工時間，進而製作出大尺寸生物支架。
　近年來，隨著醫學與科技的進步，發展出組織工程這項新的研究領域，讓受損的器官或組織可以在體外修復或重建，幫助患者延續健康。細胞是由細胞外基質所構成的網狀支架來固定其生長，本技術製作生物支架來替代細胞外基質功能，並在支架上培養纖維母細胞，分泌生長因子幫助細胞在立體結構上生長。未來希望能應用到傷口修復、黏膜重建，以及人體組織培養上。
　本論文生物支架設計以人體肝小葉實際的形狀為發想，以仿生生物支架引導細胞生長，並加入了維管束設計，幫助細胞團塊培養。未來希望配合轉錄因子及生長激素誘導方式達成體外肝小葉組織重建。
　本研究使用波長450nm的二極體雷射與780nm的飛秒雷射，經由高倍率物鏡聚焦後曝光於具生物相容性之光阻—Ormocomp及PGSA上， PGSA為本實驗團隊研發之可被酵素降解的生物相容性材料，成功以兩種材料製作出生物支架，並將纖維母細胞成功培養與支架上。

關鍵字：奈米3D微影技術、生物相容性光阻、生物支架、肝小葉

In this paper, biological scaffolds fabricated by Nano 3D Lithography (N3L) were fabricated by Two-photo Polymerization (TPP) technology. Due to the small focal spot, the long processing time, the scaffold size was limited around 50μm, and N3L via dose accumulation effect shorten the processing time to produce large-scale scaffold.
The biological scaffold in the paper was designed from the shape of the human liver lobular, with biomimetic scaffold to guide cell growth and the vascular bundle design to improve spheroids culture. In the future, we hope to cooperate with transcription factor and growth hormone to induce in vitro liver lobule tissue reconstruction.
In this study, a diode laser with a wavelength of 450 nm and a femtosecond laser at 780 nm were used to focus on the biocompatible photoresist-Ormocomp and PGSA. PGSA was developed by our team-Jane Wang’s lab. Both of material can be applied to our system successfully.

Key words: Nano-3D lithography, biocompatible photoresist, cell scaffold, hepatic lobule

摘要…………………………………………………………………………………….i
圖目錄…………………………………………………………………………….…...v
表目錄…………………………………………………………………………...…..viii
第一章 緒論 1
第二章 文獻回顧 6
2.1 生物支架 6
2.1.1傳統型生物支架 6
2.1.2雙光子技術製作之生物支架 10
2.2 光阻材料 14
2.2.1 Ormocomp 14
2.2.2 Poly(glycerol-co-sebacate) acrylate (PGSA) 16
2.3 研究動機與目的 19
第三章 研究方法 21
3.1 奈米3D微影系統 21
3.1.1 780nm-3D微影系統 21
3.1.2 450nm-3D微影系統 23
3.2 總體實驗流程 26
3.3 試片準備 28
第四章 結果與討論 32
4.1 生物相容性實驗 32
4.2 生物降解實驗 37
4.3 膠原蛋白改質實驗 39
4.4 光阻與雷射波長敏感度實驗 41
4.5 光點對焦平面與校正 42
4.6 曝光功率範圍及線寬參數實驗 43
4.7 各式柱狀結構 46
4.8 大尺寸肝小葉細胞支架 47
4.8.1 結構設計與製作 47
4.8.2 大尺寸肝小葉細胞支架培養結果 50
4.9 小尺寸肝小葉細胞支架 51
4.9.1 結構設計與製作 51
4.9.2 小尺寸肝小葉細胞支架培養結果 52
第五章 結論與未來規劃 54
參考文獻……………………………………………………………………………..55
附錄…………………………………………………………………………………..60

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