細胞於二元自組裝單層膜修飾之玻璃表面之貼附與生長行為

Abstract

隨著醫學的發展與需求，細胞於人造表面的貼附生長行為研究也愈來愈受到重視，而細胞貼附行為更是許多細胞行為的基礎，如細胞的能動性、生長與分裂。而人造表面的表面性質則是影響細胞貼附行為的重要因素之一。相較於其他常用於細胞研究的人造表面，如聚苯乙烯、聚甲基丙烯酸甲酯、纖維，自組裝單層膜 (SAMs) 能更精準的調控表面組成，進而調控表面性質。 為了提高表面改質的彈性，本研究採用兩種官能基混合修飾的玻璃基材的自組裝單層膜系統。選用的自組裝單層膜分子為3-氨基丙三甲羥矽烷和3-巰基丙基-三甲羥矽烷。在pH 7.4的環境下，將兩種分子以不同比例混合修飾玻璃表面，能得到一系列表面電位介於24 mV和-155 mV之間的表面，並以此探討表面電位對於細胞行為的影響。 研究中使用X光光電子光譜儀 (XPS) 量測自組裝單層膜表面的化學組成，並由文獻得知其表面電位。另外以光學顯微鏡、螢光顯微鏡以及掃描式電子顯微鏡觀察小鼠胚胎纖維細胞株NIH3T3在各個混合比例的自組裝單層膜上的貼附與生長行為，包含貼附量、生長量、生長速率以及細胞形貌。研究發現，由於細胞表面整體帶負電性，基材表面上NH2官能基比例愈多，表面電位愈正，細胞的貼附量較多，細胞形狀較健康；反之，基材表面上NH2官能基比例愈少，表面電位愈負，細胞的貼附量較少，細胞生長情況較差。而生長速率則主要受接觸抑制特性影響，表面電位對其影響不大。

With the development of medicine, cell behaviors on artificial surface have gained attention among researchers. Cell adhesion is central to many cell behaviors including motility, survival and differentiation. Surface properties of artificial surface are known to be one of the key factors in cell adhesion. Compared with other artificial surface usually used in cell research such as polystyrene, PMMA, and fiber, self-assembled monolayers are known for its excellent ability to modify surface. To acquire better control of surface properties, mixed functional groups of silanes are introduced to glass surface. In this study, different ratio of 3-aminopropyltrimethoxysilane and 3-mercaptopropyltrimethoxysilane are applied to glass substrate, surfaces with a series of zeta potentials from +24 to -155 mV at pH 7.4 are obtained. The surface chemical composition of binary SAMs is quantified using x-ray photoelectron spectroscopy, while that of zeta potential can be found in reference. NIH3T3 cells are chosen to culture on these binary-SAM modified glass and incubated for several different time periods. The population density, proliferation rate and shape of cells adhered on surfaces of various potentials are examined using optical microscope, fluorescence microscope and scanning electron microscope. The results showed that surface with higher positive potential promotes cell adhesion and proliferation. On the contrary, more negative the zeta potential is, worse the cells adhere and grow. And zeta potential have little effect on proliferation rate.