Animal Science Department, North Carolina State University, Raleigh, North Carolina, USA.
J Orthop Res. 2013 Mar;31(3):385-91. doi: 10.1002/jor.22239. Epub 2012 Oct 1.
Our objective was to monitor chondrocyte gene expression at 0, 3, 7, and 14 days following in vitro impaction to the articular surface of porcine patellae. Patellar facets were either axially impacted with a cylindrical impactor (25 mm/s loading rate) to a load level of 2,000 N or not impacted to serve as controls. After being placed in organ culture for 0, 3, 7, or 14 days, total RNA was isolated from full thickness cartilage slices and gene expression measured for 17 genes by quantitative real-time RT-PCR. Targeted genes included those encoding proteins involved with biological stress, inflammation, or anabolism and catabolism of cartilage extracellular matrix. Some gene expression changes were detected on the day of impaction, but most significant changes occurred at 14 days in culture. At 14 days in culture, 10 of the 17 genes were differentially expressed with col1a1 most significantly up-regulated in the impacted samples, suggesting impacted chondrocytes may have reverted to a fibroblast-like phenotype.
我们的目的是监测体外撞击猪髌骨关节面后第 0、3、7 和 14 天软骨细胞的基因表达。髌骨关节面要么用圆柱形撞击器(25mm/s 加载速率)轴向撞击至 2000N 的负载水平,要么不撞击作为对照。在器官培养 0、3、7 或 14 天后,从全层软骨片中分离总 RNA,并通过定量实时 RT-PCR 测量 17 个基因的表达。靶向基因包括编码与生物应激、炎症或软骨细胞外基质的合成代谢和分解代谢有关的蛋白质的基因。在撞击当天检测到一些基因表达的变化,但在培养的第 14 天发生了最显著的变化。在培养的第 14 天,17 个基因中有 10 个表达差异,其中 col1a1 在受影响的样本中上调最明显,表明受影响的软骨细胞可能已经恢复为成纤维细胞样表型。