在組織工程領域中，細胞接觸誘導效應有益於促進血管新生和預防疤痕組織的生成，而細胞接觸誘導效應是利用材料的地勢形貌使細胞在其表面呈規律地方向性排列，本研究係利用雷射剝蝕於具生物相容性/生物可降解性的材料，製作出各式具不同地勢寬度和深度的式樣來探討材料地勢形貌對於細胞增生與排列的影響。

此篇研究使用的高分子材料為聚酯類的poly(glycerol sebacate) PGS 和 poly(1,3-diamino-2-hydroxypropane-co-polyol sebacate)s APS，此兩種材料因具備生物可降解性且其機械性質介於與體內軟組織相似的範圍，因此適用於軟組織工程領域。

在生物相容性的測試中，對於纖維母細胞而言，APS與PGS均具備良好的生物相容性，其中又以PGS擁有較快的生長速率，整體來說，此兩種材料均適用於皮膚組織的再生。對於內皮細胞而言，APS 和 PGS 依然具備良好的生物相容性，其中又以APS勝出PGS，整體而言，此兩種材料也適用於血管組織的組織工程。

在細胞接觸誘導效應的測試中，我們測試了各式具不同地勢式樣的APS和PGS，且控制其雷射剝蝕的溝寬間距介於5-15μm。經由研究結果發現當APS和PGS表面具7μm寬和1.5μm深的剝蝕式樣時能夠使纖維母細胞擁呈現最佳的接觸誘導效應;而當APS和PGS表面具5μm寬和1.1μm深的剝蝕式樣時則夠使內皮細胞擁呈現最佳的接觸誘導效應。

同時，此研究亦利用雷射剝蝕技術製作出微流道系統來模擬人體微血管運作，研究結果發現人體血管內皮細胞能夠貼附於系統中，顯示出此系統應用於心血管組織再生工程的可能性。

我們相信利用雷射剝蝕生物可降解高分子，同時結合本研究所得到最佳的細胞接觸誘導效應於微流道系統將能夠促進血管組織的再生，並藉此更加進一步地朝器官再生的目標前進。

Up until June of 2016, there are roughly 120,000 people waiting for organ transplant in the United States. On average, less than 15% of patients on the organ transplant waiting list successfully receives a transplant surgery, indicating the serious shortage of organs for transplantation.[1] For patients that did receive a transplant, immunologic rejections remains a large challenge. Therefore, organ regeneration via tissue engineering is considered one promising alternative for patients in need.

Contact guidance refers to cell alignment on the surface of biomaterials, and is induced by the topographic patterns and difference choices of materials. In soft tissue regeneration, the alignment of cells is critical toward preventing scar formation and promoting angiogenesis. In this work, laser patterned scaffolds are created on several biodegradable/biocompatible materials with precise width and depth to illicit cell responses.

Two biodegradable polymers, poly(glycerol sebacate) (PGS), and poly(1,3-diamino-2-hydroxypropane-co-polyol sebacate)s (APS) with analogous mechanical properties to the soft tissues are synthesized and used in this work, along with polydimethylsiloxane (PDMS).

In this study, fibroblasts were observed with good cell responses toward APS and PGS, with slight preference toward PGS, including cell morphology and proliferation rate, indicating its applicability in skin regeneration. For endothelial cells, good cell-material interactions were observed on both APS and PGS, with slight higher preference toward APS, indicating their potential applicability in vascular tissue engineering.
Studies of cell responses toward laser ablated APS and PGS with 5-15μm gratings were conducted, and directional growth of fibroblasts were observed. The best patterns among tested of contact guidance of fibroblasts on microgrooved APS and PGS were observed with 7μm gratings, and 1.5 μm deep. Meanwhile, the high contact guidance of endothelial cells was observed on laser ablated APS and PGS with 5μm gratings and at 1.1μm depth.

To achieve the ultimate goal of full organs regeneration through tissue engineering, biodegradable artificial microvascular tissues for transporting oxygen and nutrients play one of the most critical roles during regeneration.

This study combined laser ablation technique with endothelial cell culture to create microfluidic devices of APS to mimic the in vitro microvasculature. Endothelial cell attachments were observed under flow-induced shear stress in physiological level in this microfluidic system.

Through studies of contact guidance and cell seeding in microfluidic systems fabricated from laser ablation, it is believed that the combination of the two is the key to vasculature regeneration. Though there are much to be done still, this work has brought the idea of full organ regeneration one step closer to realization.

Chapter 1: Introduction 9
1.1 Introduction to Tissue Engineering 9
1.2 Introduction to Biocompatible and Biodegradable Polymeric Materials 12
1.2.1 Introduction to Poly(dimethylsiloxane)(PDMS)12
1.2.2 Introduction to Poly(Glecyrol Sebacate)s (PGS)13
1.2.3 Introduction to Poly(1,3-diamino-2-hydroxypropane-co-polyol sebacates (APS) 14
1.3 Introduction to Different Cell Line and Its Use in Biomedical Devices 17
1.3.2 Introduction to Fibroblasts and its biomedical devices 17
1.3.2.1 Introduction to Fibroblasts 17
1.3.2.2 Introduction to Skin Graft 20
1.3.3 Introduction to Endothelial Cells and Its Biomedical Applications 25
1.3.3.1 Introduction to Endothelial Cells 25
1.3.3.2 Introduction to Vascular Grafts 26
1.4 Introduction to Cell Contact Guidance 28
1.4.1 Fibroblasts (HIG82) Contact Guidance 33
1.4.2 Endothelial Cell (HUVEC) Contact Guidance 35
1.5 Introduction to Surface Wettability 37
1.6 Introduction to Laser Ablation 39
1.7 Introduction to Microfluidic Developments in Vascular Research 41
Chapter 2: Experimental Design and Fabrication 43
2.1 Material Synthesis 43
2.1.1 Poly(dimethylsiloxane) (PDMS) 43
2.1.2 Poly(glycerol sebacate) (PGS) 44
2.1.3 Poly(1,3-diamino-2-hydroxypropane-co-polyol sebacate)s(APS) 44
2.1.4 Fabrication of PGS and APS film 44
2.2 Fabrication of Microgrooved and Microfluidic Devices by Laser Ablation 45
2.2.1 Fabrication of Microgrooved Device 45
2.2.2 Fabrication of Microfluidic Device 46
2.3 Alamar Blue Cell Proliferation Assay 48
2.4 Cell Contact Guidance Test 50
2.5 Microfluidic devices with endothelial cell seeding 50
2.6Cell Analysis of Fluorescent Microscopy and Scanning Electron Microscopy 51
2.6.1 Fluorescent Microscopy 51
2.6.2 Scanning Electron Microscopy of Cells on Various Topography 51
2.6.3 Scanning Electron Microscopy of Cells in Microfluidic Devices 51
2.7 Contact Angle Test 52
2.7.1 Surface Modification through Oxygen Plasma Treatments 52
Chapter 3: Result and Discussion 53
3.1 Laser Ablated Surface 53
3.2 Water Contact Angle Assays of Various Biomaterials 53
3.2.1 Biomaterials Surfaces (APS, PGS and PDMS) 54
3.2.2 Surface Modification through oxygen plasma treatment 55
3.3 Cell Proliferation and Attachment Assays between Different Biomaterials and Cell Line 57
3.3.1 Fibroblast Cell Culture (APS, PGS and PDMS) 57
3.3.2 Endothelial Cell Culture (APS, PGS and PDMS) 61
3.4 Cell Contact Guidance (with Different Width of Microgratings) 65
3.4.1 Fibroblast Cell Contact Guidance on APS 65
3.4.2 Endothelial Cell Contact Guidance on APS 69
3.5 The Characterizations of Depths in Microgrooves through Atomic Force Microscope 73
3.5.1 The Characterizations of Depths in APS Microgrooves 73
3.5.2 The Characterizations of Depths in PGS Microgrooves 75
3.6 Cell Contact Guidance on PGS 76
3.6.1 Fibroblast Cell Contact Guidance on PGS 77
3.6.2 Endothelial Cell Contact Guidance on PGS 77
3.7 Endothelial Cell Seeding in APS Microfluidic Device 79
Chapter 4: Conclusion 82
Chapter 5: Future Work 85
Chapter 6: Reference 85

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