Department of Operative Dentistry, School of Dentistry, Federal University of Pelotas, Rua Gonçalves Chaves, 457, Pelotas, RS, CEP: 96015-560, Brazil.
Clin Oral Investig. 2011 Oct;15(5):643-8. doi: 10.1007/s00784-010-0443-5. Epub 2010 Jul 10.
The aim of this study was to evaluate the effect of different concentrations of triethylene glycol dimethacrylate (TEGDMA) on the inhibition of matrix metalloproteinase 2 (MMP-2). Mouse gingival explants were cultured overnight in DMEM and the expression of secreted enzymes was analyzed by gelatin zymography in buffers containing 5 mM CaCl(2) (Tris-CaCl(2)) in 50 mM Tris-HCl buffer with the addition of TEGDMA at different concentrations (0.62%, 1.25%, 2.5%, or 5.0% (v/v)). The gelatinolytic proteinase present in the conditioned media was characterized as matrix metalloproteinase by means of specific chemical inhibition. The matrix metalloproteinases present in the conditioned media were characterized as MMP-2 by immunoprecipitation. The eletrophoretic bands were scanned and the transmittance values were analyzed. Data was plotted and submitted to linear regression to investigate MMP-2 inhibition as a function of TEGDMA concentration. Three major bands were detected in the zymographic assays. These bands were characterized as MMP-2. Zymogene (72 kDa), intermediate (66 kDa) and active forms of MMP-2 (62 kDa) were inhibited by TEGDMA in a dose-dependent way. These findings suggest that TEGDMA could inhibit MMP-2 expression even at small concentrations.
本研究旨在评估不同浓度的三甘醇二甲基丙烯酸酯 (TEGDMA) 对基质金属蛋白酶 2 (MMP-2) 抑制作用的影响。将小鼠牙龈外植体在 DMEM 中培养过夜，然后在含有 5 mM CaCl2（Tris-CaCl2）的缓冲液中通过明胶酶谱法分析分泌酶的表达，在 50 mM Tris-HCl 缓冲液中加入不同浓度的 TEGDMA（0.62%、1.25%、2.5%或 5.0%（v/v））。通过特定的化学抑制来鉴定条件培养基中存在的明胶酶为基质金属蛋白酶。通过免疫沉淀鉴定条件培养基中的基质金属蛋白酶为 MMP-2。扫描电泳条带并分析透射率值。绘制数据并提交线性回归以研究 MMP-2 抑制作用与 TEGDMA 浓度的关系。在明胶酶谱分析中检测到三个主要条带。这些条带被鉴定为 MMP-2。MMP-2 的酶原（72 kDa）、中间产物（66 kDa）和活性形式（62 kDa）均被 TEGDMA 以剂量依赖的方式抑制。这些发现表明 TEGDMA 甚至在小浓度下也能抑制 MMP-2 的表达。
