奈米級直溝地形對骨母細胞貼附形態之影響

Influence of Nanoscale Grooved Surface on Osteoblast-like Cell Morphology

丁彥中(Yen-Chung Ting)

指導教授 ： 蔡偉博

國立臺灣大學/工學院/化學工程學研究所/碩士(2006年)

https://doi.org/10.6342/NTU.2006.00732

摘要

本研究的目的在於探討材料表面的奈米結構對於細胞之貼附、定
向的影響。本實驗將骨母細胞培養在具有奈米級直溝圖案的矽晶片表面上（溝寬為90、150、250、340、500 nm），經過不同的培養時間，再利用電子顯微鏡觀察骨母細胞在各種奈米表面上貼附的情況，並使用軟體分析細胞形態。
我們發現直到四小時後，才有明顯差別。大體而言，細胞在溝寬
大於150 nm 的表面上延伸度比在溝寬90 nm 的表面高；同時細胞沿
著溝向貼附的比例，亦是在溝寬大於150 nm 的表面上較高，顯示細
胞對於寬溝的表面反應較快。不過在一天後，各種溝寬大小的表面上的細胞均已有類似的延伸度，且幾乎都沿溝向排列，彼此之間無明顯差異。此外透過共軛焦螢光顯微鏡，亦觀察到在直溝地形的表面上，細胞的F-actin 會沿溝向平行排列，同時細胞核也會沿溝排列並且形態亦稍有拉長。
另外也使用預吸附fibronectin（Fn）的直溝矽晶片培養骨母細胞。相較於無預吸附Fn 的表面，預吸附Fn 的直溝表面上細胞較不沿溝排列，形態也較不拉長，貼附面積較大，細胞核也較不沿溝排列。雖然短時間內細胞形態會受Fn 所影響，但長時間後細胞形態仍受直溝地形強烈控制。

關鍵字

骨母細胞 ； 奈米 ； 直溝 ； 地形 ； 形態

並列摘要

Cells are known to respond to topographic cues in their natural nanometrical environment. The aim of this study is to investigate the influences of surface nanotopography on cell adhesion and orientation. Osteoblast-like cells (MG63) were cultured on a series of nanogrooved silicon wafer (90, 150, 250, 340 and 500 nm in width). At specific time point, the images of cells on different surfaces were captured by scanning electron microscopy, and cell morphology was analyzed by image software.
Initially, cells adhering on surfaces with wider grooves showed more elongated shape. The proportion of cells which aligned parallel to grooves was higher on surfaces with wider grooves. Surfaces with wider grooves promote cells to appear quicker respond of elongation and alignment. F-actin and nuclei aligned with grooves on patterned surfaces. The elongation of nuclei on patterned surfaces was also observed.
Besides, osteoblast-like cells were cultured on grooved surfaces pre-adsorbed with Fibronectin (Fn). Compared to surfaces without Fn preadsorption, cells displayed larger spreading area, less elongated appearance and less isotropic alignment on grooved surfaces with Fn pre-adsorption. The proportion of aligned nuclei also decreased on surfaces with Fn preadsorption. Cell morphology could be affected enormously by nano-grooves even if grooves were coated densely with adhesion-associated protein, Fn.

並列關鍵字

osteoblast ； nanogroove ； morphology ； topography

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