在微米和奈米尺度上對各種表面進行圖案化的技術開發對於生物材料、光電設備以及組織工程等領域備受關注，而精確控制二維或三維空間中的分子取向和相對位置是目前製造多功能分層結構所面臨的挑戰。在分層微奈米結構的製作過程中普遍會使用逐層沉積技術結合軟微影技術來達到多層薄膜的設計，並且會挑選帶電荷分子(如聚電解質)來當作組裝物質，而聚電解質是一種水溶性、低毒性以及生物相容性的聚合物，可輕易通過一些條件控制來對多層膜的厚度組成或者表面特性進行調控。然而基於該製造技術上經常受到溶液橫向擴散以及圖案解析度降低的影響，導致在圖案定義過程中不容易形成高保真度的複雜化聚電解質結構。因此本研究目的在於通過自上而下（top-down）和自下而上（bottom-up）製程技術的整合，來突破堆疊組裝所產生的侷限性，藉由將可催化降解胜肽型聚電解質的胰蛋白酶（trypsin）利用共價鍵結的方式固定於微奈米結構模板上，以製作具有特定結構的催化性模板（catalytic stamp）對帶正電荷的聚離氨酸（PLL）進行多種幾何形狀的圖案化。並且通過與帶負電荷的聚麩氨酸（PGA）反覆沉積於表面結構上以形成多層薄膜。根據我們研究結果表明三維分層結構的（PLL/PGA）薄膜系統可以透過重複聚電解質自組裝堆疊以及催化性模板選擇性溶蝕的交替過程來進行構建。該製程方法可以設計大面積週期性的複雜自組裝結構，基於明確定義的異質表面系統還能實現開發多功能位點表面的潛力。目前已嘗試將微奈米級結構化的聚電解質多層膜用於提供微奈米級空間侷限來引導膠原蛋白選擇性吸附的排列行為，還能調控表面化學性質作為膠原蛋白奈米纖維圖案轉移的仿生界面。

Surface modified with polyelectrolyte multilayers (PEMs) offers diverse physico-chemical properties and potential applications. Layer-by-layer (LBL) deposition is commonly used in the construction of PEMs, but the limitation of stacking and lateral diffusion results in losing structural integrity while patterning PEMs. To overcome the limitation, an innovative method that combines top-down and bottom-up manners was developed to construct 3D hierarchical micro- and nanoscale biodegradable PEMs using selective degradation of catalytic stamp combined with LBL deposition. A patterned poly-L-lysine (PLL, positively charged PE) layer fabricated via enzymatic degradation using a designed polydimethylsiloxane (PDMS) stamp immobilized with enzyme on a substrate serves as foundation for LBL deposition of another negatively charged PEs (L-glutamic acid, PGA). 3D hierarchical PLL/PGA multilayered films can be fabricated through repeating the action of catalytic stamp degradation and the alternate electrostatic deposition of PGA and PLL. Such PLL/PGA multilayered films enable to investigate the adsorption behavior of proteins onto PE-based nanotopography. Our results demonstrate that PE-based nanotopography with various geometries not only provides the spatial organization and electrostatic adsorption of protein films but also facilitates the transfer of the protein patterns via bioconjugation chemistry.

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