胜肽改質之次微米溝脊表面對肌肉 以及韌帶細胞之生長與功能之影響

Abstract

細胞與材料表面的交互作用在組織工程的研究課題中尤為重要，許多體外研究主要在於模擬細胞在體內的週遭組織之構形來達到檢視細胞行為之目的，骨骼肌肉以及韌帶組織在人體中皆呈現束狀的線性排列，在體外研究利用次微米溝脊表面來將細胞引導成線性排列已經被證實，細胞的行為除了受到表面地形的影響以外，細胞外間質的訊號對細胞而言亦是一重要因素，在本研究中同時設計胜肽修飾表面以及次微米溝脊表面來分別代表對細胞的化學以及物理訊號，進而檢視細胞對於此兩種訊號之生長及功能表現。 在本研究中，經過含有RGD以及YIGSR序列組之胜肽改質的表面不只對骨骼肌肉細胞的生長速度與分化能力有所幫助，對細胞骨骼以及貼附蛋白的表現亦有顯著的差異，在次微米溝脊表面上所形成的成熟肌小管亦呈現與體內組織相似的線性排列，在韌帶的研究部份，經由多巴胺塗佈、以及含RGD序列組之胜肽接枝後的表面，在藉由偵測細胞數與螢光染色檢視後證明該材料對韌帶細胞的生長速度與細胞外間質分泌能力皆有所幫助，而在次微米溝脊表面的引導下，韌帶細胞的分泌能力比平坦表面上更佳，由本研究的結果指出，模擬細胞在體內的組織構形以及化學刺激，對於在體外的培養有著生長以及功能上顯著的幫助。

Cell-substrate interaction is a key point for tissue engineering, and many studies have focused on mimetic of the cell’s natural tissue environment to investigate the cellular behaviors. Both skeletal muscles and ligaments are constructed in bundles of fibers in human body, and the requirements for cell-guidance in aligned shape in vitro can be successful reached by the submicro-grooved pattern substrates. There are two approaches for cell behavior observation which are chemical signal, such as extracellular matrix (ECM) signals; and physical signals, such as topography effect, structural of substrates, and surrounding fluids. In this study, both of these two signals were designed to investigate the effect to cell proliferation and functionalities. The RGD- and YIGSR- containing PLGA patterned surfaces not only proved to enhance the proliferation and differentiation of cell but they also improved the focal adhesion and cytoskeleton presentation. The dopamine-coated and RGD-conjugated pattern surfaces also showed an enhancement of cellular proliferation and secretion of ECM content for ligament fibroblast, which was observed by cell number detection and fluorescent staining. The results indicated that both chemical signals from ECM-derivates and physical signals that mimetic the nature alignment shape of cell can successfully enhance cell proliferation and functionalities in vitro.