Haku Ken, Muramatsu Takashi, Hara Arisa, Kikuchi Akira, Hashimoto Sadamitsu, Inoue Takashi, Shimono Masaki
Department of Clinical Pathophysiology, Tokyo Dental College, Japan.
Bull Tokyo Dent Coll. 2011;52(4):173-82. doi: 10.2209/tdcpublication.52.173.
Epithelial cell rests of Malassez (ERM) are involved in the maintenance and homeostasis of the periodontal ligament. The objective of this study was to investigate the effect of mechanical stretching on cell growth, cell death and differentiation in the ERM. Cultured porcine ERM were stretched for 24 hr in cycles of 18% elongation for 1 sec followed by 1 sec relaxation. The numbers of cells and TUNEL-positive cells were then counted. The expression of mRNAs encoding gap junction protein α1 (Gja1), ameloblastin, bone morphogenetic protein 2 (BMP2), bone morphogenetic protein 4 (BMP4) and noggin were evaluated using quantitative real-time PCR. The number of cells in the stretching group was approximately 1.3-fold higher than that in the non-stretching controls at 24 hr (p<0.01). Apoptotic cells ranged from 1.9-2.5% in the stretching group at 24 hr, but were only 0.6% in the control group (p<0.01). The expression of Gja1, ameloblastin and noggin mRNAs in the stretching group was decreased at 24 hr compared with in the non-stretching group (p<0.01), whereas the expression of BMP2 and BMP4 mRNAs in the stretching group was significantly higher than that in the control group (p<0.01). Incorporation of 18 α-glycyrrhetinic acid (18GA, a gap junction inhibitor) promoted proliferation and apoptosis and confirmed both the increase of BMP2 and BMP4 and the decline of Gja1, ameloblastin and noggin in ERM. Thus, the ERM modulate cell proliferation and apoptosis, and inhibit differentiation by reducing expression of Gja1 under mechanical stretching.
马拉瑟上皮剩余(ERM)参与牙周韧带的维持和稳态。本研究的目的是调查机械拉伸对ERM中细胞生长、细胞死亡和分化的影响。将培养的猪ERM以18%伸长1秒然后1秒松弛的周期拉伸24小时。然后对细胞数量和TUNEL阳性细胞进行计数。使用定量实时PCR评估编码缝隙连接蛋白α1(Gja1)、成釉蛋白、骨形态发生蛋白2(BMP2)、骨形态发生蛋白4(BMP4)和头蛋白的mRNA表达。在24小时时,拉伸组的细胞数量比非拉伸对照组高约1.3倍(p<0.01)。在24小时时,拉伸组的凋亡细胞占1.9 - 2.5%,而对照组仅为0.6%(p<0.01)。与非拉伸组相比,拉伸组在24小时时Gja1、成釉蛋白和头蛋白mRNA的表达降低(p<0.01),而拉伸组中BMP2和BMP4 mRNA的表达明显高于对照组(p<0.01)。加入18α - 甘草次酸(18GA,一种缝隙连接抑制剂)促进了增殖和凋亡,并证实了ERM中BMP2和BMP4的增加以及Gja1、成釉蛋白和头蛋白水平的下降。因此,在机械拉伸下,ERM通过降低Gja1的表达来调节细胞增殖和凋亡,并抑制分化。