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標題: 觀察細胞於二氧化鈦奈米結構上生長的情形
Observation of Cell Growth on the Nanostructured TiO2 Surfaces
作者: 陳瑟芬
Chen, Se-Fen
Contributors: 林淑萍
Shu-Ping Lin
生醫工程研究所
關鍵字: 奈米結構;二氧化鈦;陽極氧化;表面改質;細胞生長
nanostructure;titania;anodization;surface modification;cell growth
日期: 2013
Issue Date: 2013-11-18 11:12:53 (UTC+8)
Publisher: 生醫工程研究所
摘要: 奈米技術在醫學方面已經備受關注。在本研究中,我們探討未經表面改質以及經過表面改質後的二氧化鈦奈米結構對細胞生長和型態變化的影響。首先,採用陽極氧化方法製備二氧化鈦奈米管柱,隨後利用APTMS做二氧化鈦奈米結構的表面改質。並藉由X光繞射儀和掃描式電子顯微鏡分析二氧化鈦奈米管柱結構。X光繞射儀分析顯示奈米管柱的結構為二氧化鈦,以掃描式電子顯微鏡觀察二氧化鈦奈米管柱之表面形貌,得知其管柱直徑大約為60.374nm。利用電子能譜化學分析儀分析基材鈦網、未經表面改質的二氧化鈦以及經過APTMS表面改質的二氧化鈦之特性。電子能譜化學分析儀結果顯示我們成功地在二氧化鈦表面做改質,使二氧化鈦表面帶有功能性的胺基,-NH2 binding energy為399.8eV,NH3+ binding energy為401.5 eV。為了研究奈米結構對細胞生長的影響,將3T3纖維母細胞分別培養於基材鈦網、未經表面改質的二氧化鈦以及經過APTMS表面改質的二氧化鈦的材料中,並利用顯微鏡觀察細胞生長的情形。在掃描式電子顯微鏡的觀察下,可以看見細胞健康地生長在二氧化鈦奈米結構上。另外,利用螢光顯微鏡的觀察可以看到細胞生長在二氧化鈦奈米結構上。細胞數目的定量分析上顯示出經由APTMS表面改質的二氧化鈦能夠促進細胞增生,其平均細胞數目為993,000(基材鈦網:639,000、未經表面改質的二氧化鈦:841,000)。我們發現APTMS表面改質的二氧化鈦與基材鈦網和未經改質的二氧化鈦相比, APTMS表面改質的二氧化鈦能夠提高整體細胞生長的能力達35.6%。我們的研究顯示了APTMS表面改質的二氧化鈦能夠促進細胞生長,且具有良好的生物相容性。
Nanotechnology had been drawn a lot of attentions in many biomedical aspects. In this study, we used unmodified and modified titania (TiO2) surfaces to investigate the effect of nanostructure on cell morphological change and cell growth. First, the structure of TiO2 nanotubes was prepared by anodization. After that, the nanostructures of TiO2 were further modified by 3-aminoproplytrime thexysilane (APTMS). X-ray diffraction (XRD) and scanning electron microscope (SEM) were used to analyze the structures of TiO2 nanotubes. XRD showed the nanotubes consisted of TiO2. Those TiO2 nanotubes were around 60.374 nm in diameter in SEM images. Electron spectroscopy for chemical analysis (ESCA) was used to characterize the original Ti web, unmodified TiO2, and APTMS modified TiO2. ESCA showed the successful surface modification with the specific amine (-NH2) functional groups on the surface of APTMS modified TiO2 (-NH2 was at 399.8 eV and NH3+ was at around 401.5 eV). In order to investigate the effect of TiO2 nanostructure on cell growth, 3T3 fibroblasts were independently cultured on the original Ti web, unmodified TiO2, and APTMS modified TiO2. Microscopy was used to examine the cells on the nanostructured TiO2 substrates. SEM displayed the cells healthily grew on TiO2 nanostructures. In addition, fluorescent images visualized the relative position of cells on TiO2 nanostructures. Quantitative analyses of cell numbers exhibited APTMS modified TiO2 effectively facilitated the cell proliferation with an increasingly cellular growth. We found APTMS modified TiO2 improved the overall capability of cell growth up to 35.6 % in our in-vitro observation in comparison with Ti and unmodified TiO2 substrates. Our study showed that APTMS modified TiO2 increased cell growth and possessed better biocompatibility.
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