聚二甲基硅氧烷基材表面修饰及力学强度变化对关节软骨细胞的影响

摘要

背景:聚二甲基硅氧烷因具有良好的生物相容性,优异的微纳米尺度可加工性,被广泛应用于细胞生物学的基础研究.但聚二甲基硅氧烷材料疏水性强,并不适于细胞贴附,因此表面修饰非常重要.目的:考察不同表面修饰方法并结合聚二甲基硅氧烷材料力学强度变化对牛关节软骨细胞生物学行为的影响.方法:采用3种不同方法修饰聚二甲基硅氧烷薄膜材料,包括胎牛血清孵育、胶原沉积和等离子体处理,同时变化聚二甲基硅氧烷材料的硬度,然后接种牛关节软骨细胞,并通过细胞骨架染色以及CCK-8表征分析其黏附、增殖情况,通过天狼星红/番红O染色以及糖胺聚糖定量表征分析基质分泌情况.结果与结论:①细胞黏附和增殖:细胞骨架染色以及生长曲线可以看出血清孵育、胶原沉积和等离子体处理均能显著改善牛关节软骨细胞在聚二甲基硅氧烷表面的黏附和增殖,其中等离子体处理的效果最好;②糖胺聚糖的总量:只有等离子体处理的表面显著提高了细胞分泌的糖胺聚糖的总量,但是细胞分泌糖胺聚糖的能力在所有修饰的表面上有所降低;③材料硬度对细胞黏附、生长和糖胺聚糖分泌都有影响,而且与材料表面修饰有关.④结果证实:实验利用血清孵育、胶原沉积以及等离子体处理聚二甲基硅氧烷材料,均能有效促进牛关节软骨细胞的贴附和生长,而且其中等离子体处理效果最优,与此同时聚二甲基硅氧烷基材的硬度也对细胞行为有所影响,但程度上要次于化学修饰,而且越能促进细胞生长的材料越抑制了关节软骨细胞的基质分泌能力.%BACKGROUND: Polydimethylsiloxane (PDMS) is widely used in the basic research on cell biology because of its good biocompatibility and ability to be processed at the micro/nano level. However, cell culture on PDMS has been generally compromised by its strong hydrophobicity.OBJECTIVE: To perform a comparative investigation on the influences of different surface modifications as well as stiffness of PDMS on cellular behaviors.METHODS: PDMS films with varying stiffnesses were subjected to various surface modifications, including serum incubation, type I collagen deposition and air plasma treatment. Bovine articular chondrocytes were seeded on PDMS films and cell adhesion, proliferation and matrix production were characterized using F-actin staining, cell counting kit-8,microscopic examination, Sirius red/Safranin-O staining and quantitative determination of glycosaminoglycans,respectively.RESULTS AND CONCLUSION: Serum incubation, type I collagen deposition and air plasma treatment were all found to promote adhesion and proliferation of bovine articular chondrocytes from the results of F-actin staining and cell prolifereration curve, with air plasma treatment the best. Total amount of glycosaminoglycans (GAG) secretion was only increased by air plasma treatment and GAG/DNA was decreased by all modifications. Stiffness also played roles in cell adhesion, proliferation and GAG production, which was found to be dependent on surface modifications. This study would provide guidance for applying PDMS in cell culture.