細胞外基質（extracellular matrix ，ECM）可通過微米及奈米尺度之表面形態調節細胞行為，這代表有機會利用材料表面的結構影響源祖細胞(progenitor cell)的生長與分化，進一步可能達到以生醫材料控制幹細胞分化的目的，為達到此此一目標，對於材料表面起伏結構與幹細胞表現間關係的研究是必需的。本研究利用軟蝕刻（soft lithography）技術，將微溝槽轉印在二甲基矽氧烷（polydimethalsiloxane，PDMS）薄膜上，微溝槽的寬度由1µm 至 20 µm，深度則維持於0.8µm，並藉由氬離子電漿，將疏水的二甲基矽氧烷改質為親水。本研究利用兩種能顯示成骨細胞分化的細胞；類骨母細胞（osteoblast-like cells）及牙髓幹細胞（dental pulp stem cells）培養於溝槽表面，針對細胞的排列性（alignment）、延展性（elongation）、貼附（attachment）、延伸（spreading）、增生（proliferation）、細胞骨架分佈（cytoskeleton distribution）及細胞分化（cell differentiation）進行分析。
實驗結果顯示，牙髓幹細胞與類骨母細胞的排列性與延展性明顯隨著溝槽寬度減少而增加，這指出細胞在溝槽上具有接觸引導（contact guidance）的現象。此外，藉由細胞骨架免疫染色顯示基動蛋白組織纖維結構，發現細胞雙極結構所產生的偽足及基動蛋白絲具有高度方向性會順著溝槽方向生長。除了密度外基動蛋白絲方向性也會隨溝槽寬度變窄而增加。當溝槽寬度為1-13µm時，溝槽結構會阻礙牙髓幹細胞貼附、延展和增生；反之，此類表面型態卻會促進類骨母細胞的貼附、延展及增生，這一截然不同的影響可能與細胞本身的延展性或大小相關。此外，關於幹細胞的分化表現，實驗結果指出隨著溝槽寬度下降，骨分化(ALPase)表現會增加，且CD44的表現明顯降低，這證實了微溝槽有助於細胞之骨分化表現。本研究指出微溝槽表面會促進牙髓幹細胞的分化及方向性。此外，材料表面的潤濕性也會影響材料表面結構對細胞的調節作用，即因為細胞親和力的增加，親水性表面能增加細胞對為溝槽表面的辨識效果，因而在親水性的表面上，細胞因微溝槽出現的貼附、延展及排列行為將更加明顯。

The extracellular matrix (ECM) regulates cells’ behaviors via a lot of ways, including the surface topography in micro- or nano-scale. It is proposed that the topography of biomaterials may be applied to control the differentiation of multipotent cells, which is especially important for stem cells. In this research, the microgrooved topography was transferred to the polydimethylsiloxane (PDMS) film by using the soft lithography method. The width of grooves and ridges varied from 1 µm to 20 µm, while the groove depth was kept about 0.8 µm. By exposing to Argon plasma treatment, PDMS surface changed from hydrophobic into hydrophilic. The two kinds of cells were cultured on the grooved surfaces in this research, including osteoblast-like cells and multipotent cells which may express osteoblastic differentiation. For dental pulp stem cells (DPSCs), cell alignment, elongation, attachment, spreading, proliferation, cytoskeleton distribution, and cell differentiation were then analyzed. The cell alignment and elongation were significantly enhanced by microgroove patterns and increased with the decrease in the groove width. It indicated that “contact guidance” existed. In addition, the organization of actin filaments was visualized by the cytoskeleton immunostaining. On the grooved surface, cells adopted bipolar morphology with few lamellipodia and expressed highly orientated actin filaments along the groove direction. The density as well as orientation degree of actin filaments would be increased with the narrowing of groove width. On the other hand, cells exhibited meshwork of peripheral actin filament with many lamellipodia when they were cultured on flat surface. The cell attachment, spreading, proliferation were retarded by the small groove patterns (1-13 µmm) despite this structure could offer a larger specific area. In contrast, groove topography promoted the attachment, spreading, and proliferation of osteoblast-like cells. Furthermore, early osteoblastic differentiation (ALPase) of stem cells was notably enhanced corresponding to the reduction of groove width. In addition, CD44 expression obviously decreased by decreasing groove width. This finding confirms that narrow groove patterns are able to promote osteoblastic differentiation of DPSCs. The research outcomes implicate that microgrooved surfaces would promote the orientation and then the differentiation of multipotent cells. On the other hand, the influences of surface wettability were also examined and the results indicate that hydrophilic surface could enhance the effect of groove patterns. Because of the high affinity between cells and hydrophilic surfaces, cultured cells would recognize topographical cues better on grooved PDMS with high wettability.

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