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  • 學位論文

不同基材的軟硬度對皮膚細胞的影響

Effect of substratum stiffness on skin cell behavior

指導教授 : 林頌然

摘要


組織工程有三大要素:一為細胞(cell),二為支撐細胞生長所需的支架(scaffold),三為影響細胞行為的訊息因子(signal)。三大要素之間會有互相影響作用,兩兩之間的的交互作用也是組織工程的重要課題。 細胞和培養細胞的基材(支架)交互作用影響很深遠,已經知道基材的機械性質可以調控細胞的行為。本實驗想了解基材的軟硬度對皮膚細胞行為的影響。我們利用真皮乳頭細胞、纖維母細胞及表皮角質細胞,利用不同軟硬度的polydimethylsiloxane(PDMS)為基材,來看對皮膚細胞的影響。 實驗的結果顯示纖維母細胞的貼附、型態及增生,幾乎不受到基材軟硬度影響而有所變化,這代表纖維母細胞在不同基材軟硬度上的環境適應性良好。真皮乳頭細胞在越硬的基材上細胞的貼附度較好,不過細胞增生能力不受到軟硬度的影響。在不同基材軟硬度上面,真皮乳頭細胞皆表現其獨有的基因表現。真皮乳頭細胞在不同軟硬度的基材上,且都展現細胞聚集的行為。先前的研究顯示真皮乳頭細胞聚集時會有比較好的誘導毛囊再生的能力,因此PDMS很適合用以當作維持真皮乳頭細胞毛囊誘導能力的培養基材。表皮角質細胞的貼附度會隨硬度增加而增加,而增生速率則在較軟的基材上有比較好的表現。其細胞型態沒有明顯受到基材軟硬之影響,角質細胞在不同基材的軟硬度上都可以進行終端分化。在角質分化相關的基因表現量來看,在越軟的基材上面角質細胞往終端分化的基因表現量是越高的。本研究顯示,不同的皮膚細胞,對於基材的軟硬度感受不同。因此對於不同的皮膚細胞,可藉由選定特定基材的軟硬度,來達到期望的細胞培養效果。

並列摘要


There are three elements of tissue engineering: cell, scaffold, and signal. These elements inteact with each other to modulate the final cell behavior. It has been shown that the mechanical property of the substratum material can regulate cells. Here, we explore how substratum stiffness affects skin cell behavior. We focus on three skin cell types: dermal papilla cells, fibroblasts, and keratinocytes. We use polydimethylsiloxane (PDMS) of different stiffness for cell culture. Fibroblasts grow well on substratum of different stiffness. They show a similar trend of attachment, proliferation, and cell morphology despite the substratum stiffness is varied. Dermal papilla cells adhere better on stiffer substratum. They tend to self-aggregate and preserve specific makers on substratum of different stiffness. Keratinocyte are more adhesive to stiffer substratum, but they proliferate faster on softer substratum. However, their differentiation is not significantly affected by the substratum stiffness and cells can reach final differentiation on PDMS substratum with different stiffness. The results suggest that substratum stiffness affects differentially on different cell types. The substratum stiffness should be tailored for the specific purpose when different skin cell types are cultured.

參考文獻


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