Matsui Satoshi, Takahashi Chitaka, Tsujimoto Yasuhisa, Matsushima Kiyoshi
Department of Endodontics, Nihon University School of Dentistry at Matsudo, Matsudo, Chiba, Japan.
J Endod. 2009 Jan;35(1):67-72. doi: 10.1016/j.joen.2008.08.034.
The present study was conducted to investigate the effects of reactive oxygen species (ROS) on the calcification ability of human dental pulp (HDP) cells. HDP cells were treated with 100 mumol/L hydrogen peroxide (H(2)O(2)) for 5 or 10 minutes (5-min ROS group and 10-min ROS group) to investigate the mechanism of transmission to cells. Untreated cells were used as controls. Generation of free radicals was quantified by the electron spin resonance spin-trapping method and found to be increased by treatment with ROS. Formation of calcified nodules was also investigated by von Kossa staining and alizarin red S staining. Twenty-eight days after exposure, calcified nodules were present in cell cultures that had been treated with ROS for 5 or 10 minutes. Expression of mRNAs for osteopontin (OPN) and osteocalcin (OCN) was significantly greater in 10-min ROS group 6 and 9 days, respectively, after exposure than in controls. Production of OPN and OCN by 10-min ROS group was also greater 12 and 18 days, respectively, after exposure than in controls. These results suggested that calcification of HDP cells was stimulated by H(2)O(2) and by the ROS it generated.
本研究旨在探讨活性氧（ROS）对人牙髓（HDP）细胞钙化能力的影响。将HDP细胞用100μmol/L过氧化氢（H₂O₂）处理5或10分钟（5分钟ROS组和10分钟ROS组），以研究其向细胞内传递的机制。未处理的细胞用作对照。通过电子自旋共振自旋捕获法对自由基的产生进行定量，发现ROS处理后自由基产生增加。还用冯科萨染色法和茜素红S染色法研究了钙化结节的形成。暴露28天后，在经ROS处理5或10分钟的细胞培养物中出现了钙化结节。在暴露后第6天和第9天，10分钟ROS组中骨桥蛋白（OPN）和骨钙素（OCN）的mRNA表达分别显著高于对照组。在暴露后第12天和第18天，10分钟ROS组中OPN和OCN的产生也分别高于对照组。这些结果表明，HDP细胞的钙化受到H₂O₂及其产生的ROS的刺激。
