鈦表面經電化學處理後對不同組織特性的類造骨細胞貼附、增殖及分化之影響

Effect of electrochemical treatment of the titanium surface on different histotypic features osteoblast-like cells adhesion, proliferation and differentiation

莊羽喬(Yu-Chiao Chuang)

指導教授 ： 李天翎 ； 黃何雄

中山醫學大學/口腔醫學院/口腔材料科學研究所/碩士(2006年)

摘要

本研究之目的在探討於純鈦金屬表面進行電化學陽極處理後其生物相容性質的好壞。本實驗中試片先以砂紙研磨至#1500（#1500：號數為1500號的砂紙），接著再於試片表面進行電化學陽極處理，外加陽極電流使試片表面形成一層奈米網狀結構的氧化層，接著以體外細胞培養技術探討此奈米網狀結構的生物相容性質。以場發射掃描式電子顯微鏡觀察材料表面的奈米網狀結構。將兩種不同組織特性的類造骨細胞（U-2 OS、MG-63）培養於試片上以分析生物相容性質。使用各種生醫技術評估細胞在鈦試片上的反應，例如：細胞的貼附、增殖及分化。以場發射掃描式電子顯微鏡觀察細胞的形態。類造骨細胞鹼性磷酸酶活性的表現即為細胞分化的標誌。接著再以von Kossa染色觀察細胞於試片上鈣化的情形。結果顯示，隨著截止電流的增加，材料表面網目大小也會變大。兩種不同組織特性的細胞，測出的增殖速度也有所差異。細胞貼附形態方面則可發現具有奈米網狀結構的試片表面細胞貼附的情形是比較好的。不同條件的試片表面細胞鹼性磷酸酶活性的表現皆有些許差異。U-2 OS的高峰點出現在一星期，而MG-63的高峰點則是出現在第四天，這或許是細胞組織特性的關係才會有這樣的差異。具有網狀結構的試片表面細胞的鹼性磷酸酶活性會有較高的表現量。總而言之，以電化學陽極處理備製出奈米網狀結構的材料表面會具有良好的生物相容性。

關鍵字

鈦 ； 電化學 ； 奈米網狀結構 ； 類造骨細胞 ； 生物相容性 ； 鹼性磷酸酶活性

並列摘要

The specific aim of the present study was to investigate the biocompatibility of pure titanium surface after electrochemical anodization treatment. In this study, the titanium surface were polished to #1500 by sandpaper, prepared the network-structured oxide layer on pure titanium substrate using the electrochemical anodization treatment, and then in vitro cell culture technologies were used to investigate the biocompatibility of the surface modified titanium. Surface characterizations of the network-structured layer was analyzed using field emission scanning electron microscope (FE-SEM). For biocompatibility, two cell lines of osteoblast-like cells (U-2 OS、MG-63) with different histotypic features were cultured on the titanium specimens. The cell responses, e.g. cell adhesion, cell proliferation and cell differentiation, on the titanium specimens were evaluated using various biomedical techniques. The morphology of the attached cells on specimens was observed using FE-SEM. The initial differentiation of osteoblast-like cells was evaluated as a function of alkaline phosphatase activity. The calcification of osteoblast-like cells was observed using von Kossa stain on titanium specimens. The results showed the network size increased on titanium surface with an increase in the applied anodic current. Different speed of cell proliferation was measured from two kinds osteoblast-like cells with different histotypic features. In cell adhesion morphology, the cells have the best adhesion morphology were those attached on nano-network surface. There are some differences in alkaline phosphatase activity of osteoblast-like cells grew on different titanium specimens surface. U-2 OS alkaline phosphatase peaked activity at week 1, MG-63 alkaline phosphatase peaked activity at day 4, this may result from the various histotypic feature that come with different cell lines. Cells on nano-network surface has higher alkaline phosphatase activity than the other surface. Altogether, nano-network surface with a better biocompatibility can be prepared on pure titanium substrate using an electrochemical anodization treatment.

並列關鍵字

Titanium ； Electrochemical ； Nano-network ； Biocompatibility ； Osteoblast-like cells ； ALP activity

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