Mikulás Krisztina, Komlódi Timea, Földes Anna, Sváb Gergely, Horváth Gergő, Nagy Ádám Miklós, Ambrus Attila, Gyulai-Gaál Szabolcs, Gera István, Hermann Péter, Varga Gábor, Tretter László
Department of Prosthodontics, Semmelweis University, 1088 Budapest, Hungary.
Department of Medical Biochemistry, MTA-SE, Laboratory of Neurobiochemistry, Semmelweis University, 1094 Budapest, Hungary.
Materials (Basel). 2020 Aug 6;13(16):3472. doi: 10.3390/ma13163472.
Triethylene glycol dimethacrylate (TEGDMA) monomers released from resin matrix are toxic to dental pulp cells, induce apoptosis, oxidative stress and decrease viability. Recently, mitochondrial complex I (CI) was identified as a potential target of TEGDMA. In isolated mitochondria supported by CI, substrates oxidation and ATP synthesis were inhibited, reactive oxygen species production was stimulated. Contrary to that, respiratory Complex II was not impaired by TEGDMA. The beneficial effects of electron carrier compound methylene blue (MB) are proven in many disease models where mitochondrial involvement has been detected. In the present study, the bioenergetic effects of MB on TEGDMA-treated isolated mitochondria and on human dental pulp stem cells (DPSC) were analyzed.
Isolated mitochondria and DPSC were acutely exposed to low millimolar concentrations of TEGDMA and 2 μM concentration of MB. Mitochondrial and cellular respiration and glycolytic flux were measured by high resolution respirometry and by Seahorse XF extracellular analyzer. Mitochondrial membrane potential was measured fluorimetrically.
MB partially restored the mitochondrial oxidation, rescued membrane potential in isolated mitochondria and significantly increased the impaired cellular O consumption in the presence of TEGDMA.
MB is able to protect against TEGDMA-induced CI damage, and might provide protective effects in resin monomer exposed cells.
从树脂基质中释放出来的二甲基丙烯酸三甘醇酯(TEGDMA)单体对牙髓细胞有毒性,可诱导细胞凋亡、氧化应激并降低细胞活力。最近,线粒体复合物I(CI)被确定为TEGDMA的一个潜在靶点。在由CI支持的分离线粒体中,底物氧化和ATP合成受到抑制,活性氧生成增加。与此相反,呼吸复合物II未受到TEGDMA的损害。电子载体化合物亚甲蓝(MB)的有益作用在许多检测到线粒体参与的疾病模型中得到了证实。在本研究中,分析了MB对TEGDMA处理的分离线粒体和人牙髓干细胞(DPSC)的生物能量学效应。
将分离的线粒体和DPSC急性暴露于低毫摩尔浓度的TEGDMA和2μM浓度的MB中。通过高分辨率呼吸测定法和海马XF细胞外分析仪测量线粒体和细胞呼吸以及糖酵解通量。用荧光法测量线粒体膜电位。
在存在TEGDMA的情况下,MB部分恢复了线粒体氧化,挽救了分离线粒体中的膜电位,并显著增加了受损的细胞氧消耗。
MB能够防止TEGDMA诱导的CI损伤,并可能在暴露于树脂单体的细胞中提供保护作用。