Atsumi T, Iwakura I, Fujisawa S, Ueha T
Department of Oral Physiology, School of Dentistry, Meikai University, Sakado-Shi, Saitama, Japan.
Biomaterials. 2001 Jun;22(12):1459-66. doi: 10.1016/s0142-9612(00)00267-2.
In order to clarify the mechanism of photo-damage caused by eugenol (4-allyl-2-methoxyphenol), we measured cell survival in the presence of eugenol at concentrations of 10(-3) - 10(-7) M, with and without VL (visible light) irradiation by a VL dental lamp and at various pHs (7.2, 7.8 and 8.2) using two different cells (HSG, a human submandibular gland tumor cell line; HGF, a human gingival fibroblast in primary culture). Also, ROS (reactive oxygen species) generation in the above adherent single cells was measured by ACAS laser cytometry combined with CDFH-DA, a peroxide probe. The survival of both HSG and HGF cells treated with eugenol was significantly decreased as the VL irradiation time and/or the pH of the medium was increased. The amount of ROS generated from eugenol was also enhanced by increasing the VL irradiation time and elevating the pH of the medium. Cytotoxicity and ROS generation of HGF cells were significantly lower than that of HSG cells. Glutathione (1 mM) or cysteine (1 mM) protected the photo damages. We conclude that the cytotoxicity of VL-irradiated eugenol possibly was caused by the generation of eugenol radicals and additionally by ROS, both of which were produced dependent on the dose of eugenol, length of irradiation time, and pH of the medium.
为了阐明丁香酚（4-烯丙基-2-甲氧基苯酚）引起光损伤的机制，我们使用两种不同的细胞（HSG，一种人下颌下腺肿瘤细胞系；HGF，一种原代培养的人牙龈成纤维细胞），在丁香酚浓度为10(-3)-10(-7)M的情况下，分别在有和没有VL（可见光）照射（使用VL牙科灯）以及在不同pH值（7.2、7.8和8.2）条件下测量细胞存活率。此外，通过结合过氧化物探针CDFH-DA的ACAS激光细胞术测量上述贴壁单细胞中活性氧（ROS）的生成。随着VL照射时间和/或培养基pH值的增加，用丁香酚处理的HSG和HGF细胞的存活率均显著降低。随着VL照射时间的增加和培养基pH值的升高，丁香酚产生的ROS量也增加。HGF细胞的细胞毒性和ROS生成显著低于HSG细胞。谷胱甘肽（1 mM）或半胱氨酸（1 mM）可保护细胞免受光损伤。我们得出结论，VL照射的丁香酚的细胞毒性可能是由丁香酚自由基的产生以及另外由ROS引起的，这两者的产生均取决于丁香酚的剂量、照射时间长度和培养基的pH值。
