本論文主要利用超快雷射之非線性多光子激發技術行光交聯(photocrosslinking)反應於蛋白質溶液中，聚合加工出二維與三維生醫材料結構，所使用的加工溶液包含孟加拉玫瑰素(rose Bengal，RB)作為光活化劑(photoactivator)，以牛血清白蛋白(bovine serum albumin，BSA)或第一型膠原蛋白(type I collagen)為反應單體。孟加拉玫瑰素經雙光子激發後可有效的產生高活性的單態氧，此單態氧會與蛋白質氨基酸的可氧化端產生反應並形成活化的蛋白質，而該蛋白質將可與另一個蛋白質反應，形成鍵結並完成雙光子交聯作用。該反應機制經由實驗室所發展之超快雷射加工系統，可加工出具有生物相容性(biocompatible)的三維結構。另外也可藉由聲光調變器(acousto-optic modulator，AOM)調控每個加工點雷射光源之脈衝數量，以達到不同聚合程度的灰階加工。論文中也嘗試利用此蛋白聚合結構以物理性方式限制及引導膠原蛋白纖維化方向，並對其纖維化程度利用外加氯化鈣(calcium chloride)方式進一步做些調整，期望使其可均勻分布於結構內；於第一型膠原蛋白實驗中，藉以中和膠原蛋白溶液使孟加拉玫瑰素保有其功能下，成功地直接對膠原蛋白聚合加工出二維大面積結構。這些蛋白質加工結構的研究可擴展應用於複雜細胞外基質(extracellular matrix，ECM)製作與組織工程(tissue engineering)等研究。

In this thesis, nonlinear multiphoton excited technique is utilized to fabricate two-/three-dimensional (2D/3D) structures via photocrosslinking in protein solution. The solution contains photoactivator, rose Bengal (RB), and reactive monomers, bovine serum albumin (BSA) or type I collagen. RB with two-photon excitation can efficiently cause surrounding oxygen into singlet oxygen. The singlet oxygen then reacts with the oxidable species of amino acid and the protein is induced as high activity. Further, this protein reacts with another protein and form a chemical bond. The reaction process by using a home-made femtosecond laser processing system can deliver biocompatible 3D structures. Besides, different gray level laser dose has been adopted to create complex biomaterial structures via laser pulse number modulation for different fabrication regions. The protein fabricated structure is also employed to physically constrain and guide collagen fibril direction. The degree of collagen fibril is also modulated by adding calcium chloride for leading the fibril uniformly distributed in fabricated microstructures. In addition, a large area 2D collagen structure has been successfully fabricated by regulating collagen solution to neutral condition and keeping the photoactivator ability of RB. The study of protein fabricated structure can further apply to complex extracellular matrix (ECM) and tissue engineering aspects.

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