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软骨细胞在纳米结构微柱聚丙烯和聚苯乙烯表面的行为。

Chondrocyte behavior on nanostructured micropillar polypropylene and polystyrene surfaces.

作者信息

Prittinen Juha, Jiang Yu, Ylärinne Janne H, Pakkanen Tapani A, Lammi Mikko J, Qu Chengjuan

机构信息

Department of Applied Physics, University of Eastern Finland, Kuopio, Finland.

Department of Chemistry, University of Eastern Finland, Joensuu, Finland.

出版信息

Mater Sci Eng C Mater Biol Appl. 2014 Oct;43:424-31. doi: 10.1016/j.msec.2014.07.045. Epub 2014 Jul 19.

Abstract

This study was aimed to investigate whether patterned polypropylene (PP) or polystyrene (PS) could enhance the chondrocytes' extracellular matrix (ECM) production and phenotype maintenance. Bovine primary chondrocytes were cultured on smooth PP and PS, as well as on nanostructured micropillar PP (patterned PP) and PS (patterned PS) for 2 weeks. Subsequently, the samples were collected for fluorescein diacetate-based cell viability tests, for immunocytochemical assays of types I and II collagen, actin and vinculin, for scanning electronic microscopic analysis of cell morphology and distribution, and for gene expression assays of Sox9, aggrecan, procollagen α1(II), procollagen α1(X), and procollagen α2(I) using quantitative RT-PCR assays. After two weeks of culture, the bovine primary chondrocytes had attached on both patterned PP and PS, while practically no adhesion was observed on smooth PP. However, the best adhesion of the cells was on smooth PS. The cells, which attached on patterned PP and PS surfaces synthesized types I and II collagen. The chondrocytes' morphology was extended, and an abundant ECM network formed around the attached chondrocytes on both patterned PP and PS. Upon passaging, no significant differences on the chondrocyte-specific gene expression were observed, although the highest expression level of aggrecan was observed on the patterned PS in passage 1 chondrocytes, and the expression level of procollagen α1(II) appeared to decrease in passaged chondrocytes. However, the expressions of procollagen α2(I) were increased in all passaged cell cultures. In conclusion, the bovine primary chondrocytes could be grown on patterned PS and PP surfaces, and they produced extracellular matrix network around the adhered cells. However, neither the patterned PS nor PP could prevent the dedifferentiation of chondrocytes.

摘要

本研究旨在探究图案化聚丙烯(PP)或聚苯乙烯(PS)是否能增强软骨细胞的细胞外基质(ECM)生成及表型维持。将牛原代软骨细胞培养在光滑的PP和PS上,以及纳米结构微柱PP(图案化PP)和PS(图案化PS)上,培养2周。随后,收集样本进行基于荧光素二乙酸酯的细胞活力测试、I型和II型胶原蛋白、肌动蛋白和纽蛋白的免疫细胞化学分析、细胞形态和分布的扫描电子显微镜分析,以及使用定量逆转录聚合酶链反应(RT-PCR)分析Sox9、聚集蛋白聚糖、前胶原α1(II)、前胶原α1(X)和前胶原α2(I)的基因表达。培养两周后,牛原代软骨细胞附着在图案化PP和PS上,而在光滑PP上几乎未观察到附着。然而,细胞在光滑PS上的附着最佳。附着在图案化PP和PS表面的细胞合成了I型和II型胶原蛋白。软骨细胞形态伸展,在图案化PP和PS上附着的软骨细胞周围形成了丰富的ECM网络。传代时,尽管在第1代传代软骨细胞的图案化PS上观察到聚集蛋白聚糖的最高表达水平,且传代软骨细胞中前胶原α1(II)的表达水平似乎降低,但在软骨细胞特异性基因表达上未观察到显著差异。然而,所有传代细胞培养物中前胶原α2(I)的表达均增加。总之,牛原代软骨细胞可在图案化PS和PP表面生长,并在附着细胞周围产生细胞外基质网络。然而,图案化PS和PP均不能阻止软骨细胞的去分化。

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