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研究生：	林郁庭 Lin, Yu-Ting
論文名稱：	三維組織培養用多孔雙電性水凝膠之製備 Synthesis of Porous Polyampholyte Hydrogel Scaffold for 3-D Tissue Culture
指導教授：	劉瑞祥 Liu, Jui-Hsiang
學位類別：	碩士 Master
系所名稱：	工學院 - 化學工程學系 Department of Chemical Engineering
論文出版年：	2020
畢業學年度：	108
語文別：	英文
論文頁數：	88
中文關鍵詞：	雙電性、水凝膠、多孔支架、細胞培養、動態離子鍵
外文關鍵詞：	polyampholyte, hydrogel, porous scaffold, cell culture, dynamic ionic bonding
相關次數：	點閱：54 下載：0
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通過動態離子鍵和永久性共價鍵的形成，成功構建了聚兩性電解質水凝膠。聚兩性電解質是由分佈無規則的陽離子和陰離子重複基團所組成，其離子鍵所形成的動態網絡為水凝膠提供了強大的機械性能，同時促進細胞在水凝膠中的遷移。水凝膠中還另外摻入含羥基的HEMA單體以改善含水量，此非離子單體會改變超分子結構，顯著影響所製備凝膠的物理性質。
微結構研究結果表明，水凝膠中為高密度多孔結構，其平均孔徑為72-103 μm，此孔徑大小可增加生物分子的擴散效率。由適當配方所製備的水凝膠在生理條件下具有高含水量，穩定的熱性質和合適的機械性質。生物實驗的結果表明，PA/HEMA比例為 60 / 40、80 / 20和100/0水凝膠顯現出出色的細胞附著性，這歸因於水凝膠網絡內的靜電相互作用。其中，在80/20水凝膠中培養的NHDF和L929細胞保持較高的增殖能力和生存率，表明了PA水凝膠作為細胞外基質取代物的可行性。

The polyampholyte hydrogel scaffolds were constructed successfully through the formation of dynamic ionic linkages and permanent covalent linkages. Polyampholytes are composed of randomly dispersed cationic and anionic repeating groups. The dynamic ionic network gives the hydrogels strong mechanical properties and promotes cell migration in the scaffold. HEMA containing hydroxyl group is incorporated to improve water content. Introducing of non-ionic units changes supramolecular construction and significantly affects the properties of the fabricated gels.
The results of morphology study showed high density porous structure with average pore size of 72-103 μm, which improved biomolecule diffusion obviously. With proper formulations, the hydrogels possessed high water content, stable thermal properties and suitable mechanical properties under physiological conditions. The PA/HEMA-60/40, 80/20, and 100/0 hydrogels showed great cell attachment performance dependent on covalent and electrostatic interactions within the hydrogel network. Among them, the NHDF and L929 cells cultured on the PA/HEMA-80/20 hydrogel scaffolds showed a high viability and proliferative capacity, revealing that the PA hydrogels could provide possibility for extracellular matrix replacement.

Contents
Abstract	I
中文摘要	II
Contents	III
List of Tables	VI
List of Figures	VII
1.	Introduction	1
1-1	Preface	1
1-2	Research motivation	2
2.	Literature Review	5
2-1	Introduction of Hydrogel	5
2-2	Classification of the Hydrogels	7
2-2-1	Physical and chemical hydrogels	9
2-2-2	Supramolecular polymeric hydrogels	13
2-3	Application of Hydrogels	22
2-4	Hydrogels for Tissue Engineering	30
3.	Experiment	34
3-1	Materials	34
3-2	Instruments	35
3-3	Experimental section	36
3-3-1	Synthesis of Polyampholyte (PA) Hydrogel Scaffolds	36
3-3-2	Sample preparation for compressive test	40
3-3-3	SEM sample preparation	40
3-3-4	Solid State 13C-NMR Spectroscopy (SSNMR)	41
3-3-5	Measurement of Equilibrium Water Content (EWC)	41
3-3-6	Thermogravimetric analysis (TGA) for PA/HEMA hydrogels	42
3-3-7	Cell Culture and Proliferation	42
3-3-8	Adhesion Test and Biocompatibility Test of Polyampholytes Hydrogels	43
3-3-9	Immunostaining analysis	45
4.	Results and Discussion	47
4-1	Characterization of the Hydrogel Scaffolds	47
4-1-1	Components Ratio of PA/HEMA Hydrogel Scaffolds	47
4-1-2	Structure Identification	48
4-1-3	Thermal Properties of the Hydrogels	52
4-1-4	Microstructure and Morphology of the Hydrogels	53
4-1-5	Water Content of Various Polyampholyte Hydrogels	60
4-1-6	Mechanical Behavior of the Hydrogels	62
4-2	Hydrogel as Extracellular Matrix Mimics for Cell Culture	65
4-2-1	Cell morphology	65
4-2-2	In Vitro Cell Adhesion Test	67
4-2-3	In Vitro Cell proliferation Test	72
4-2-4	In Vitro Cell Viability Test	76
5.	Conclusions	82
References	83


                                    

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