本論文研究主要在探討機械性質與表面親疏水特性對類骨細胞(UMR-106)生理行為表現之影響，本實驗系統中以聚二甲基矽氧烷(PDMS, SYLGARD 184TM)為培養基材原料，藉著調控寡聚物與硬化劑之間的重量比例，製備出不同機械強度的基材，並在特定的參數控制下以氬氣(Argon)低溫電漿改質的方式，將原為疏水性的基材表面改質成適合細胞貼附生長的親水性基材表面，並進行一系列的材料強度、表面特性與細胞生理行為分析和觀察，其中以拉伸測試作材料強度分析，而表面特性有靜態表面接觸角和傅立葉轉換-遠紅外線分析，細胞行為反應分析則包含了在有無血清培養環境中的細胞貼附率(Cell attachment)、細胞增殖(Cell proliferation)、細胞移動速度(Cell migration)、細胞延展(Cell spreading)、細胞延伸率(cell elongation)、細胞鹼性磷酸酶表現(Cell Alkaline Phosphatase )和細胞的鈣化行為(Cell mineralization)。

在本實驗系統中成功地藉由控制寡聚物與硬化劑的重量比例從10：1.2至10：0.3，所得之彈性模數從最硬約2200KPa至最軟約80KPa，且經由分析證實，機械強度的差異並未對材料的表面親疏水特性和化學結構造成大幅度的改變。除此之外，本實驗所控制的氬氣電漿處理參數亦可成功地明顯改變材料表面的潤濕特性(接觸角度從約102o變成約45o)，且未造成表面的化學結構和機械強度明顯的改變。

在細胞生理反應的實驗觀察中，在含有血清的培養條件下，在電漿改質過的親水性表面材料上有較佳的細胞貼附表現，機械強度的影響也較為明顯，因為此表面有較佳的親和性，骨細胞能在短時間內進行貼附，進而清楚的辨識機械強度，並引導細胞的生理行為。比較不同機械強度的材料，硬材料能有較佳的誘導效果，因此能有效刺激骨細胞內的化學訊號傳遞鏈，然而在未改質過的疏水性表面的細胞貼附行為中，因為表面缺乏親和性，因此機械強度並無任何明顯的效應。

然而在細胞的增生表現中，不管是親水性或疏水性表面，硬材料皆不利於細胞的增生行為，因此可認為不同生長階段的細胞貼附與增生對機械強度有不同的效應和需求，也就是說硬材料能促進細胞的初期貼附，但卻可能不利於細胞的後期增生行為，除此之外，細胞行為中的延展面積、鹼性磷酸酶(ALP)和礦化表現行為，亦以硬材料有較佳的誘導細胞內訊號的傳遞，因此有較佳的表現。而其中細胞的移動行為則以在親和性較弱的的疏水性表面有較快的滑動速度，且硬材料上則能有效促進形成偽足而有明顯具目標性的移動。

然而在不含血清的培養系統中，因為失去周圍蛋白質的影響，細胞表現出明顯的軸向拉伸行為外，其中細胞的初期貼附、面積延展亦皆以在親合性較佳的硬材料上有較佳的辨識和誘導，然而細胞的移動行為卻因蛋白質效應的消失而失去對細胞的引導，而降低了骨細胞在任何材料上的移動速度。

本論文的目標在於期望能夠獲得有關於機械效應誘導細胞行為的訊息，並能在生醫材料的設計上和控制細胞反應的行為中能有所貢獻，因此由本研究中指出，材料的機械特性和表面親疏水性質對刺激骨細胞的生理行為，如貼附、增生和分化，確實有明顯的效應存在，並以接觸角約45o親水性表面且強度約2200kPa的硬材料能有效刺激骨細胞分化的生理表現，且根據本研究，不同的細胞生長時期和環境會對不同的機械強度產生反應，因此在未來有關骨再生的生醫產業對於特性設計和材料的選擇上，本研究將會是值得參考一大重點，以期能有效控制骨細胞的生理反應。

The main purpose of this work is the investigation of osteoblastic cells’ (UMR-106) phenotypes on surfaces with different mechanical properties. The PDMS (polydimethyl siloxane) substrates with different elastic moduli by controlling the ratio of oligomer to crosslinker were prepared and applied for the culture of osteoblastic cells. Meanwhile, the hydrophilicity of substrates was also adjusted by using the argon plasma treatment. The elastic modulus was determined from the tensile test, and the surface characteristics were evaluated by contact angle measurement and FTIR analysis. Finally, the behaviors of osteoblastic cells were analyzed, including attachment, proliferation, migration, spreading, elongation, alkaline phosphatase (ALPase) expression and mineralization performance.

By controlling the percentage of crosslinkers, we can successfully adjust the elastic modulus of PDMS substrates from 2200KPa to 80KPa. However, the wettability and surface functional groups were not obviously changed with the mechanical strength. On the other hand, the plasma treatment would decrease the contact angle of PDMS substrates (from 102o to 45o); meanwhile, it did not result in significant difference in the mechanical strength and the chemical composition of substrates.

With the presence of serum, the cell attachment on the hydrophilic and hard PDMS substrate would be the best and the influence of mechanical strength would very significant. Because of the good affinity between cells and this hydrophilic surface, the osteoblastic cells can attach onto the substrate in a short time and recognize the mechanical strength. Thus, the mechanical strength would affect the cell behavior immediatedly. For the comparison of different mechanical strength, the UMR-106 would prefer to attach to the hard substrate which may stimulate the signal chain inside the cells. For the cell attachment on the hydrophobic pristine substrate, there would be no differences caused by the variations of mechanical strength. It was because that the cells can not recognize the surface properties immediately due to the low cell-surface affinity.

However, long-term proliferation was not good on the hardest surfaces., no matter on hydrophilic or hydrophobic surfaces. It suggested that the requirements of mechanical strength would be different for different stages, cell attachment and proliferation. In the initial stage of cell attachment, hard surface would be beneficial to support cells. On the contrary, the surface which was too stiff would not be suitable for cell proliferation. The cell spreading, expression of alkaline phosphatase (ALPase) and mineralization would be all better on the hardest PDMS, which was probably due to the activation in cellular signaling. The cell mobility would be higher on the hydrophobic substrate with poor cell affinity, especial on the hard substrate. Hard surface would promote formation of lamellopodia, which was necessary for cell migration.

In the the system without serum, the cells would be elongated along a single axis. Besides, the cell mobility and spreading would be decreased, compared with the system added with serum. The effect of mechanical strength would be briefly the same with those in the system with serum addition; that was, the hard surface would be beneficial to the cell attachment, mobility and spreading.

The aim of this thesis is to obtain useful information of mechanical strength for the design of medical devices and the control of cell responses. Thus, the present research revealed that substrate elasticity and surface hydrophilicity can significantly affect the attachment, proliferation and differentiation of osteoblast-like cells. According to the stages of cells and surroundings, the different mechanical strength could be selected according to this research.

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