Baldión Paula Alejandra, Díaz Carlos Aldemar, Betancourt Diego Enrique
Departamento de Salud Oral, Facultad de Odontología, Sede Bogotá, Universidad Nacional de Colombia, Bogotá, Colombia.
J Biomed Mater Res A. 2025 Feb;113(2):e37872. doi: 10.1002/jbm.a.37872.
The activity of matrix metalloproteinases (MMPs) plays a crucial role in the aging of the resin-dentin interface. The in situ action of MMP-2 and MMP-9 has been confirmed in the process of dentin-collagen degradation. However, the involvement of dental pulp cells in MMP secretion as a response to oxidative stress induced by contact with resin monomers has not been fully elucidated. Myricetin (MYR), like proanthocyanidin (PAC), has antioxidant properties and may help prevent extracellular matrix degradation. The objective was to evaluate the effect of MYR on the MMP expression and activity in response to reactive oxygen species (ROS) increase induced by triethylene glycol dimethacrylate (TEGDMA) in human odontoblast-like cells (hOLCs). hOLCs differentiated from dental pulp mesenchymal stem cells were exposed to TEGDMA released from dentin blocks using a barrier model with transwell inserts for 18, 24, and 36 h. Intracellular oxidation was evaluated using the 2',7'-dichlorofluorescein probe. The effect of 600 μM MYR on the MMP-2 and MMP-9 expression was determined by reverse transcription-quantitative polymerase chain reaction (RT-qPCR). The extracellular MMP levels were quantified using enzyme-linked immunosorbent assay, and their activation by means of a proteolytic fluorometric assay. The results were analyzed by one-way analysis of variance and Tukey's post hoc test, p ≤ 0.05. TEGDMA exposure increased intracellular ROS and upregulated MMP-2 and MMP-9 mRNA in hOLCs (p < 0.001). The levels of MMPs increased significantly 24 h after TEGDMA exposure (p = 0.013). These secreted proteases exhibited high activation ability. MYR reduced ROS production and downregulated MMP expression and activity at both mRNA and protein levels, similar to the effect found for PAC, which was used as a control. A relationship was observed between MMP-2 and MMP-9 expression, secretion, and early activation with ROS increase due to TEGDMA exposure. MYR showed potential as a therapeutic strategy to control MMP expression and modulate redox imbalance, offering a protective effect on cellular response.
基质金属蛋白酶（MMPs）的活性在树脂-牙本质界面的老化过程中起着关键作用。MMP-2和MMP-9在牙本质胶原降解过程中的原位作用已得到证实。然而，牙髓细胞作为对与树脂单体接触诱导的氧化应激的反应而参与MMP分泌的情况尚未完全阐明。杨梅素（MYR）与原花青素（PAC）一样，具有抗氧化特性，可能有助于防止细胞外基质降解。目的是评估MYR对人成牙本质细胞样细胞（hOLCs）中因二缩三乙二醇二甲基丙烯酸酯（TEGDMA）诱导的活性氧（ROS）增加而导致的MMP表达和活性的影响。使用带有transwell小室的屏障模型，将从牙髓间充质干细胞分化而来的hOLCs暴露于牙本质块释放的TEGDMA中18、24和36小时。使用2',7'-二氯荧光素探针评估细胞内氧化。通过逆转录-定量聚合酶链反应（RT-qPCR）确定600μM MYR对MMP-2和MMP-9表达的影响。使用酶联免疫吸附测定法定量细胞外MMP水平，并通过蛋白水解荧光测定法测定其活性。结果通过单因素方差分析和Tukey事后检验进行分析，p≤0.05。TEGDMA暴露增加了hOLCs中的细胞内ROS，并上调了MMP-2和MMP-9 mRNA（p<0.001）。TEGDMA暴露24小时后，MMP水平显著增加（p=0.013）。这些分泌的蛋白酶表现出高激活能力。与用作对照的PAC的效果相似，MYR在mRNA和蛋白质水平上均降低了ROS产生，并下调了MMP表达和活性。观察到MMP-2和MMP-9的表达、分泌以及早期激活与TEGDMA暴露导致的ROS增加之间存在关联。MYR显示出作为控制MMP表达和调节氧化还原失衡的治疗策略的潜力，对细胞反应具有保护作用。
