Department of Oral Cell Biology, Academic Centre for Dentistry Amsterdam (ACTA), University of Amsterdam and VU University Amsterdam, Research Institute MOVE, Amsterdam, The Netherlands.
J Cell Biochem. 2010 May;110(1):260-71. doi: 10.1002/jcb.22533.
The extracellular matrix surrounding chondrocytes within a chondron is likely to affect the metabolic activity of these cells. In this study we investigated this by analyzing protein synthesis by intact chondrons obtained from different types of cartilage and compared this with chondrocytes. Chondrons and chondrocytes from goats from different cartilage sources (articular cartilage, nucleus pulposus, and annulus fibrosus) were cultured for 0, 7, 18, and 25 days in alginate beads. Real-time polymerase chain reaction analyses indicated that the gene expression of Col2a1 was consistently higher by the chondrons compared with the chondrocytes and the Col1a1 gene expression was consistently lower. Western blotting revealed that Type II collagen extracted from the chondrons was cross-linked. No Type I collagen could be extracted. The amount of proteoglycans was higher for the chondrons from articular cartilage and nucleus pulposus compared with the chondrocytes, but no differences were found between chondrons and chondrocytes from annulus fibrosus. The expression of both Mmp2 and Mmp9 was higher by the chondrocytes from articular cartilage and nucleus pulposus compared with the chondrons, whereas no differences were found with the annulus fibrosus cells. Gene expression of Mmp13 increased strongly by the chondrocytes (>50-fold), but not by the chondrons. Taken together, our data suggest that preserving the pericellular matrix has a positive effect on cell-induced cartilage production.
软骨陷窝周围的细胞外基质可能会影响软骨细胞的代谢活性。在这项研究中,我们通过分析从不同类型软骨获得的完整软骨陷窝的蛋白质合成来研究这一点,并将其与软骨细胞进行比较。来自不同软骨来源(关节软骨、髓核和纤维环)的山羊软骨陷窝和软骨细胞在藻酸盐珠中培养 0、7、18 和 25 天。实时聚合酶链反应分析表明,与软骨细胞相比,软骨陷窝的 Col2a1 基因表达始终更高,而 Col1a1 基因表达始终更低。Western blot 显示从软骨陷窝提取的 II 型胶原发生交联。未提取到 I 型胶原。与软骨细胞相比,来自关节软骨和髓核的软骨陷窝的蛋白聚糖含量更高,但纤维环的软骨陷窝和软骨细胞之间没有差异。与来自纤维环的细胞相比,来自关节软骨和髓核的软骨细胞的 Mmp2 和 Mmp9 表达更高,而软骨陷窝则没有差异。软骨细胞的 Mmp13 基因表达强烈增加(>50 倍),但软骨陷窝没有。总的来说,我们的数据表明,保留细胞外基质对细胞诱导的软骨生成有积极影响。