Faculty of Dentistry, National University of Singapore, Singapore.
J Periodontal Res. 2012 Apr;47(2):212-21. doi: 10.1111/j.1600-0765.2011.01423.x. Epub 2011 Oct 20.
Cell adhesion plays important roles in maintaining the structural integrity of connective tissues and sensing changes in the biomechanical environment of cells. The objective of the present investigation was to extend our understanding of the effect of cyclic mechanical strain on the expression of adhesion-related genes by human periodontal ligament cells.
Cultured periodontal ligament cells were subjected to a cyclic in-plane tensile deformation of 12% for 5 s (0.2 Hz) every 90 s for 6-24 h in a Flexercell FX-4000 Strain Unit. The following parameters were measured: (i) cell viability by the MTT assay; (ii) caspase-3 and -7 activity; and (iii) the expression of 84 genes encoding adhesion-related molecules using real-time RT-PCR microarrays.
Mechanical stress reduced the metabolic activity of deformed cells at 6 h, and caspase-3 and -7 activity at 6 and 12 h. Seventy-three genes were detected at critical threshold values < 35. Fifteen showed a significant change in relative expression: five cell adhesion molecules (ICAM1, ITGA3, ITGA6, ITGA8 and NCAM1), three collagen α-chains (COL6A1, COL8A1 and COL11A1), four MMPs (ADAMTS1, MMP8, MMP11 and MMP15), plus CTGF, SPP1 and VTN. Four genes were upregulated (ADAMTS1, CTGF, ICAM1 and SPP1) and 11 downregulated, with the range extending from a 1.76-fold induction of SPP1 at 12 h to a 2.49-fold downregulation of COL11A1 at 24 h.
The study has identified several mechanoresponsive adhesion-related genes, and shown that onset of mechanical stress was followed by a transient reduction in overall cellular activity, including the expression of two apoptosis 'executioner' caspases.
细胞黏附在维持结缔组织的结构完整性和感知细胞生物力学环境变化方面发挥着重要作用。本研究旨在深入了解周期性机械应变对人牙周韧带细胞中黏附相关基因表达的影响。
在 Flexercell FX-4000 应变单元中,培养的牙周韧带细胞每 90 秒接受 12%的平面内拉伸变形 5 秒(0.2 Hz),持续 6-24 小时。测量以下参数:(i)MTT 测定法测定细胞活力;(ii)半胱天冬酶-3 和 -7 活性;(iii)使用实时 RT-PCR 微阵列检测 84 种编码黏附相关分子的基因的表达。
机械应力在 6 小时降低了变形细胞的代谢活性,在 6 和 12 小时降低了半胱天冬酶-3 和 -7 的活性。在临界阈值 <35 时检测到 73 个基因。15 个基因的相对表达发生显著变化:5 个细胞黏附分子(ICAM1、ITGA3、ITGA6、ITGA8 和 NCAM1)、3 个胶原α链(COL6A1、COL8A1 和 COL11A1)、4 个 MMPs(ADAMTS1、MMP8、MMP11 和 MMP15),加上 CTGF、SPP1 和 VTN。4 个基因上调(ADAMTS1、CTGF、ICAM1 和 SPP1),11 个基因下调,范围从 12 小时 SPP1 的 1.76 倍诱导到 24 小时 COL11A1 的 2.49 倍下调。
本研究鉴定了几个机械反应性黏附相关基因,并表明机械应激的起始伴随着整体细胞活性的短暂降低,包括两种凋亡“执行者”半胱天冬酶的表达。