- 學位論文
波浪狀電紡絲藉張力增進韌帶基因表現
Crimped Electrospun Fibers Upregulate Ligamentogenic Gene Expression Via a Tension-Mediated Pathway
摘要
波浪狀的電紡絲支架已經被研究為韌帶組織工程的仿生環境。波浪狀的微環境已被證實能增進韌帶基因的特性與支架的機械強度，但是波浪狀纖維對於細胞表型的分子機制尚未被研究。 在這個研究中，前十字韌帶纖維母細胞被種在波浪狀纖維支架上。波浪狀纖維調控了細胞形態與細胞骨架結構，這意味著細胞張力的變化。肌球蛋白在肌動蛋白中扮演聚合與重組的作用，所以此研究調查了磷酸化肌球蛋白在波浪狀細胞的細胞反應中有何作用。我們的結果表明，在波浪狀支架上的細胞有較高的肌球蛋白表現。當加入會降低肌球蛋白磷酸化程度的ROCK抑制劑Y27632或MLCK抑制劑ML-7，在直線與波浪狀纖維的細胞韌帶基因表現沒有顯著差異。這些結果意味著前十字韌帶纖維母細胞的肌球蛋白與細胞張力負責調控波浪狀細胞的表現效果。
並列摘要
Electrospun fibrous crimped scaffolds have been studied as a biomimetic environment for ligament tissue engineering. Crimped microenvironments have been shown to improve ligamentogenic properties and scaffold mechanical strength, but the molecular mechanism of fiber crimp on cell phenotype has not yet been investigated. In this study, ACL fibroblasts were seeded on fibrous crimped scaffolds. Fiber crimp modulates cell morphology and cytoskeletal structure, implying a change in cell tension. Myosin plays a role in actin polymerization and reorganization, so this study investigates the role of myosin phosphorylation in the cellular response to a crimped morphology. Our results showed that cells seeded on crimped scaffolds had higher levels of myosin expression. When treated with ROCK inhibitor Y27632 or MLCK inhibitor ML-7, which reduce the levels of myosin phosphorylation, ligamentogenic gene expression of cells on straight and crimped fibers had no significant differences. These results imply that myosin and cell tension are responsible for mediating the effect of crimp morphology on ACL fibroblasts.