Department of Oral Biology, Goldschleger School of Dental Medicine, Tel-Aviv University, Tel-Aviv, Israel.
J Periodontal Res. 2010 Apr;45(2):200-6. doi: 10.1111/j.1600-0765.2009.01218.x. Epub 2009 Nov 9.
Periodontal disease is characterized by increased expression and activity of matrix metalloproteinases (MMPs) and insufficient expression/activity of their inhibitors, tissue inhibitors of matrix metalloproteinases (TIMPs). This altered MMP-TIMP balance results in progressive destruction of gingival and periodontal extracellular matrix. Enamel matrix derivative (EMD), clinically used for periodontal regeneration in a device called Emdogain, has been suggested to enhance gingival healing following periodontal procedures in humans. We previously showed that EMD increases the proliferation of human and rat gingival fibroblasts and protects them from tumor necrosis factor-induced apoptosis. In the present study, the modulation of MMP and TIMP expression by EMD was investigated.
Primary human gingival fibroblasts were treated in vitro with tumor necrosis factor, EMD or both in serum-free conditions, and RNA was analyzed with an extracellular matrix-focused microarray and quantitative real-time polymerase chain reaction.
Microarray analysis showed detectable expression of MMP-1, MMP-2, MMP-3, MMP-7 and MMP-13, as well as TIMP-1 and TIMP-3 in untreated cells. There was no apparent regulation of the expression of MMP-2, MMP-7, MMP-13 and TIMP-1 by either tumor necrosis factor or EMD. In contrast, tumor necrosis factor significantly increased MMP-1 expression, and EMD reduced it when both agents were present. Also, EMD significantly induced TIMP-3 expression, an effect which was dependent on activation of extracellular signal-regulated kinase 1/2, since it was totally abolished by a selective extracellular signal-regulated kinase pathway inhibitor.
These data suggest that EMD may affect gingival health by ways other than cell proliferation/survival, i.e. by stimulation of TIMP-3 production, which could improve the MMP-TIMP balance in gingival tissue and curb extracellular matrix destruction.
牙周病的特征为基质金属蛋白酶（MMPs）表达和活性增加，以及其抑制剂，基质金属蛋白酶抑制剂（TIMP）的表达和活性不足。这种 MMP-TIMP 平衡的改变导致牙龈和牙周细胞外基质的进行性破坏。釉基质衍生物（EMD），临床上用于一种名为 Emdogain 的装置中的牙周再生，已被证明可增强牙周手术后人类牙龈的愈合。我们之前表明，EMD 可增加人牙龈成纤维细胞的增殖并保护其免受肿瘤坏死因子诱导的凋亡。在本研究中，研究了 EMD 对 MMP 和 TIMP 表达的调节。
体外用人肿瘤坏死因子、EMD 或两者在无血清条件下处理原代人牙龈成纤维细胞，并通过细胞外基质聚焦微阵列和实时定量聚合酶链反应分析 RNA。
微阵列分析显示，未经处理的细胞中可检测到 MMP-1、MMP-2、MMP-3、MMP-7 和 MMP-13 以及 TIMP-1 和 TIMP-3 的表达。肿瘤坏死因子或 EMD 均未明显调节 MMP-2、MMP-7、MMP-13 和 TIMP-1 的表达。相反，肿瘤坏死因子显著增加了 MMP-1 的表达，而当两种药物同时存在时，EMD 则降低了它的表达。此外，EMD 还显著诱导了 TIMP-3 的表达，这种作用依赖于细胞外信号调节激酶 1/2 的激活，因为它被一种选择性的细胞外信号调节激酶途径抑制剂完全阻断。
这些数据表明，EMD 可能通过刺激 TIMP-3 产生来影响牙龈健康，而不是通过细胞增殖/存活，这可以改善牙龈组织中的 MMP-TIMP 平衡并抑制细胞外基质的破坏。
