自组装膜表面不同功能团对骨骼肌细胞生物学特性的影响

Effect of different functional groups on self-assembled monolayers on the biological characteristics of skeletal muscle cells in vitro

目的探讨自组装膜表面不同的功能团对骨骼肌细胞生物学特性影响。方法通过自组装技术在金片表面接枝不同的化学功能团(-CH3、-COOH、-NH2、-OH),测定接触角和原子力显微镜确定其表征,将材料与骨骼肌细胞进行共培养,分别通过SEM、DAPI染色、MTT法、钙黄绿素染色和流式检测法等方法观察不同功能团对骨骼肌细胞的形态、粘附、增殖、活力及凋亡的影响。结果含有-NH2和-COOH基团的材料表面能明显促进骨骼肌的粘附、增殖,具有良好的生物相容性;而含有-CH3基团的基片明显不利于骨骼肌细胞的粘附、增殖,并对骨骼肌细胞凋亡有一定的促进;不同的功能团材料对骨骼肌细胞粘附、增殖率影响的顺序从大到小依次为-NH2>-COOH>-OH>-CH3;不同的化学功能团对细胞凋亡率和毒性的顺序-NH2<-COOH<-OH<-CH3。结论不同的功能团对骨骼肌细胞的生物特性具有不同的影响,含有-NH2基团的基片能明显促进细胞的粘附和增殖,有利于构建具有促细胞粘附的生物材料界面;而含有-CH3基团的材料不利于细胞的粘附和增殖,有利于构建抗粘附的生物材料表面界面。

Objective To explore the effect of different functional groups on self- assembled monolayers on the biological characteristics of rabbit skeletal muscle cells in vitro. Methods Rabbit skeletal muscle cells were cultured on self- assembled monolayers of gold on which different terminal chemical groups including methyl groups（- CH3）, amino（- NH2）, hydroxyl（-OH） and carboxyl（- COOH） were anchored with self- assembled methods. Contact angle measurements and atomic force microscopy were employed to confirm the similar density of different functional groups occupation. Fluorescence microscopy,MTT assay, flow cytometry, and scanning electron microscopy（SEM） were used to analyze the morphological and biological alterations of the cells. Results SEM results revealed that the chemical groups on the surface of the monolayer modulated the structure of skeletal muscle cells and the cell morphology. Skeletal muscle cells cultured on the monolayer with-CH3 exhibited the smallest contact area with a spherical morphology, while the cells on the monolayers with-NH2,-OH and-COOH showed much larger contact area and flatter morphology. The functional groups-NH2and-COOH obviously promoted cell adhesion and proliferation, while-CH3 group produced significantly greater toxicity than-NH2,-OH and-COOH groups to inhibit the cell growth and adhesion and promote cell death. Cell attachment and growth was enhanced, in the order the magnitude of the effect, by-NH2-COOH-OH-CH3, and the toxicity decreased in the order of-NH2-COOH-OH-CH3. Conclusion The terminal chemical groups can obviously affect the phenotype of skeletal muscle cells in vitro, and this finding provides a theoretical basis for surface design of biomaterials.