超音波具方向性之能量與作用力傳遞,因此其微流動作用( micro-streaming )可分為水平方向和垂直方向,然而平行流動板僅單一水平方向。本實驗分兩個部份一部分採用超音波之作用力照射,模擬微血管內之環境,建構出水平與垂直方向之超音波微流場,使細胞接受40o、70o、90o不同角度超音波刺激,並觀察細胞遷移速度的差異。第二部分利用密閉之平行流動板,建構一水平機械力流場,觀察細胞遷移速度的差異。最後再測細胞存活率、細胞排序角度與免疫螢光染色,觀察細胞骨架 actin 、細胞貼附蛋白 vinculin及細胞核。結果顯示:超音波刺激對細胞遷移影響顯著,其中又以間接刺激的影響最為顯著;在細胞排序數據發現,超音波直接刺激40o細胞排序角度最小;平行流動板的刺激在細胞排序的影響力較不顯著;透過免疫螢光染色發現超音波間接刺激70o的vinculin分布最多,而間接與直接刺激90o對細胞造成些微傷害使細胞型態改變。
Ultrasound is a transfer of energy and force, which can be divided into horizontal and vertical direction. The parallel plate flow chamber can form a horizontal flow field. Cell migration is the fundamental cellular process for normal development and homeostasis of a tissue and organ. In this study, 40°, 70° and 90° of ultrasonic stimulation were applied on the human gingival fibroblast cells. The parallel plate flow chamber constructed a horizontal flow field. Then, the speed of cell migration was observed. The cell survival ability, alignment angle analysis, immunofluorescence staining of cytoskeleton, vinculin, and nucleus were evaluated. Slant ultrasonic stimulation remarkable influence the cell migration. The data of alignment angle analysis showed that the 40o ultrasonic direct stimulates was the smallest. The parallel plate flow chamber stimulation displayed inconsistent alignment angle between cultured cells. 70° ultrasonic indirect stimulates by the displayed marked vinculin in the immunofluorescence. However, 90° ultrasonic stimulates display slightly cell injured.
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