Orthopaedic Research Laboratory, Aarhus University Hospital, Aarhus, Denmark.
Interdisciplinary Nanoscience Center, iNANO, University Aarhus, Aarhus, Denmark.
J Biomed Mater Res A. 2018 Nov;106(11):2805-2816. doi: 10.1002/jbm.a.36467. Epub 2018 Aug 27.
Understanding the behavior of chondrocytes in contact with artificial culture surfaces is becoming increasingly important in attaining appropriate ex vivo culture conditions of chondrocytes in cartilage regeneration. Chondrocyte transplantation-based cartilage repair requires efficiently expanded chondrocytes, and the culture surface plays an important role in guiding the behavior of the cell. Micro- and nano-engineered surfaces make it possible to modulate cell behavior. We hypothesized that the combined influence of topography, substrate, and surface chemistry may affect the chondrocyte culturing in terms of proliferation and phenotypic means. Human chondrocytes were cultured on polystyrene fabricated microstructures, flat polydimethylsiloxane (PDMS), or polystyrene treated with fibronectin or oxygen plasma and cultured for 1, 4, 7, and 10 days. The behavior of chondrocytes was evaluated by proliferation, viability, chondrogenic gene expression, and cell morphology. Contrary to our hypothesis, microstructures in polystyrene did not significantly influence the behavior of chondrocytes neither under normoxic- nor hypoxic conditions. However, changes in the substrate stiffness and surface chemistry were found to influence cell viability, gene expression, and morphology of human chondrocytes. Oxygen plasma treatment was the most important parameter followed by the softer substrate type PDMS. The findings indicate the culture of human chondrocytes on softer substratum and surface activation by oxygen plasma may prevent dedifferentiation and may improve chondrocyte transplantation-based cartilage repair. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 106A: 2805-2816, 2018.
了解软骨细胞在与人工培养表面接触时的行为,对于获得软骨再生中软骨细胞的适当体外培养条件变得越来越重要。基于软骨细胞移植的软骨修复需要有效扩增的软骨细胞,而培养表面在指导细胞行为方面起着重要作用。微纳工程表面使调节细胞行为成为可能。我们假设,形貌、基底和表面化学的综合影响可能会影响细胞培养的增殖和表型方式。人软骨细胞在聚苯乙烯制成的微结构、平整的聚二甲基硅氧烷(PDMS)或用纤维连接蛋白或氧等离子体处理的聚苯乙烯上培养,并分别培养 1、4、7 和 10 天。通过增殖、活力、软骨生成基因表达和细胞形态来评估软骨细胞的行为。与我们的假设相反,在常氧和低氧条件下,聚苯乙烯中的微结构都不会显著影响软骨细胞的行为。然而,基底硬度和表面化学的变化被发现会影响人软骨细胞的活力、基因表达和形态。氧等离子体处理是最重要的参数,其次是较软的 PDMS 基底类型。研究结果表明,在较软的基质上培养人软骨细胞并通过氧等离子体进行表面激活,可能防止去分化,并可能改善基于软骨细胞移植的软骨修复。© 2018 Wiley Periodicals, Inc. J 生物材料 Res 部分 A:106A:2805-2816,2018。