||Zirconia coating on 316L stainless steel prepared by sol-gel method and assessed by interacting with vascular endothelial cells
||Department of Materials Science and Engineering
vascular endothelial cells
本研究利用溶膠凝膠法，嘗試在低溫製程下(< 400 °C)在醫用316L不鏽鋼上製備出氧化鋯鍍層，探討不同退火溫度、溶液組成、鍍層層數等參數對物性、化性及機械性質的影響，分析係藉由掃描式電子顯微鏡以及原子力顯微鏡分析薄膜表面形貌；以X光光電子能譜儀分析薄膜表面化學組成；並使用奈米壓痕試驗機的側向力及刮痕模組探討奈米尺度鍍層抵抗側向施力的能力。最後與人類臍靜脈內皮細胞共同培養以評估材料的生物相容性。經24、48小時候使用免疫螢光染色觀察細胞貼附形貌，並使用MTT assay分析在不同材料上細胞的增生活性，最後搭配酵素免疫分析法評估不同材料的促發炎潛在性。
In this research, sol-gel process was utilized to prepare ceramic coating on medical grade 316L stainless steel and different experimental parameters’ influence on physical, chemical and mechanical properties, such as annealing temperature, different solution constitute and coating layers were discussed. Coating characterizations were done by AFM and SEM (surface topography and morphology), XPS (surface chemical structures) and Nanoindentation-lateral force and scratch mode. Then human umbilical vein endothelial cells were cultured on specimens to evaluate their biocompatibility, with fluorescence staining to observe the morphology of cells, and MTT assay to evaluate the proliferation activity of cells on different materials. Finally, the proinflammatory response of cells on different specimens was assessed by enzyme-linked immunosorbent assay (ELISA) in order to see its potential for the final use in biomedical applications.
Results showed that annealing is needed to remove the solvents and additives out, and to improve mechanical properties. But its main drawback is the decreasing of surface coverage.
According to the bio-evaluation results, HUVECs didn’t show significant difference on proliferation activity on ZrO2 coatings compared to 316L stainless steel, but they do show difference on proinflammation response test. This means that ceramic coating is indeed less aggressive than stainless steel to cells, which show potential for future application in surface treatment of stainless steel as for scalpels and indwelling needle.
List of Contents V
List of Tables VIII
List of Figures IX
Chapter 1. Introduction 1
1.1 Introduction 1
1.2 Motivation 2
1.3 Literature Survey 4
1.3.1 Biomedical applications of zirconia 4
1.3.2 Utilizing sol-gel method to coat zirconia on stainless steel 5
1.3.3 The influence of oxide coatings on biocompatibility 9
1.4 Objective 12
Chapter 2. Theoretical Basis 13
2.1 Sol-gel method 13
2.1.1 Principle of sol-gel method 13
2.1.2 Factors affecting sol-gel method 14
2.1.3 Introduction of sol-gel coating processes 15
2.1.4 Advantages and disadvantages of sol-gel process 16
2.2 Interfacial adhesion of thin films/coatings 17
2.2.1 Interfacial adhesion 18
2.2.2 Adhesion measurement 19
2.2.3 Scratch test 20
2.3 Bioceramics 22
2.3.1 Classes of biomaterials 22
2.3.2 Zirconia 23
2.4 Inflammation and vascular endothelial cells 25
2.4.1 Vascular endothelial cells 26
2.4.2 Role of vascular endothelial cells in inflammation 26
2.4.3 Inflammatory Cells 27
2.4.4 Different kinds of inflammation 29
2.4.5 Chemical Mediators of Inflammation 29
Chapter 3. Material and Method 31
3.1 Experimental setup 32
3.1.1 Substrate 32
3.1.2 Zirconia coating prepared by sol-gel process 32
3.2 Specimen Preparation Instruments 33
3.2.1 Spin coater 33
3.2.2 Muffle Furnace 34
3.3 Characterization instruments 34
3.3.1 Grazing Incident X-ray diffraction (GIXRD) 34
3.3.2 Differential Scanning Calorimetry (DSC) 35
3.3.3 α-step profilometer 36
3.3.4 X-ray Photoelectron Spectroscopy (XPS) 37
3.3.5 Scanning Electron Microscopy (SEM) 38
3.3.6 Atomic Force Microscopy (AFM) 39
3.3.7 Nano indenter 39
3.4 Cell culture test 42
3.4.1 Reagents and chemicals used in cell culture test 42
3.4.2 Cell culture 43
3.4.3 Method and methodology 43
3.5 Statistical analysis 45
Chapter 4. Results and Discussions 46
4.1 Sol-gel zirconia coating preparation 46
4.2 Microstructure analysis by XRD 47
4.3 Thermal analysis by DSC 50
4.4 Surface morphology and coating thickness 52
4.5 Surface chemical analysis by XPS 56
4.5.1 The influence of post-annealing 57
4.5.2The influence of polymer addition 60
4.6 Nano-Scratch test 63
4.6.1The influence of heat treatment 64
4.6.2 The influence of thickness 69
4.6.3 The influence of polymer additives 69
4.7 In vitro biocompatibility assessments 70
4.7.1 Cell morphology observation 70
4.7.2 Cell proliferation activity 71
4.7.3 Pro-inflammation response 73
Chapter 5. Conclusion 76
Appendix A. JCPD card 84
Appendix B. Additional information from cell culture tests 86
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