Tsutsui T, Matsudo Y, Tanaka Y, Rokukawa A, Sato H, Omori N, Barrett J C
Department of Pharmacology, The Nippon Dental University, School of Dentistry at Tokyo, 1-9-20, Fujimi, Chiyoda-ku, Tokyo 102, Japan.
Toxicol In Vitro. 1994 Dec;8(6):1253-8. doi: 10.1016/0887-2333(94)90117-1.
The cytotoxicities of antiseptics used in dentistry were examined with normal human keratinocytes from the oral cavity by the uptake of neutral red (NR assay). The NR assay is a quantitative in vitro assay that distinguishes between viable, damaged or dead cells. Cultures from different individuals were established, and secondary cultures in serum-free medium were used. The amount of NR extracted from cells after a 3-hr uptake of the dye was linearly proportional to the number of cells exposed to the dye. The cytotoxicity of antiseptics used on mucous membranes in the oral cavity was determined from the dose-response curves of inhibition of NR uptake in cells treated with antiseptics for 2 days. As a quantitative measure of cytotoxicity, NR(50) (i.e. the concentration of antiseptics that resulted in a 50% decrease in NR uptake relative to untreated controls) was extrapolated from dose-response curves. The rank-order of cytotoxicities (NR(50)) was Hyamine (10% benzethonium chloride solution) > Hibitane (5% chlorhexidine gluconate solution) > acrinol (0.2% acrinol solution) > Iodine Carbol (liquefied phenol with 3% iodine) > iodine glycerin (glycerin with 10% iodine) > dilute iodine tincture (3% iodine solution) approximately iodine tincture (6% iodine solution) approximately Isodine (10% povidone-iodine solution) > Oxydol (3% hydrogen peroxide solution). These results provide useful estimates of relative toxicities of antiseptics to human cells, which can be compared with antimicro-organism activity to maximize therapeutic benefit with minimal toxicity.
通过中性红摄取法（NR 测定法），使用来自口腔的正常人角质形成细胞检测了牙科用防腐剂的细胞毒性。NR 测定法是一种定量体外测定法，可区分活细胞、受损细胞或死细胞。建立了来自不同个体的培养物，并使用无血清培养基中的传代培养物。染料摄取 3 小时后从细胞中提取的 NR 量与暴露于染料的细胞数量呈线性比例。根据用防腐剂处理 2 天的细胞中 NR 摄取抑制的剂量反应曲线，确定了口腔粘膜用防腐剂的细胞毒性。作为细胞毒性的定量指标，从剂量反应曲线中推断出 NR(50)（即相对于未处理对照导致 NR 摄取减少 50%的防腐剂浓度）。细胞毒性（NR(50)）的排序为：海明（10%苄索氯铵溶液）> 洗必泰（5%葡萄糖酸氯己定溶液）> 吖啶黄（0.2%吖啶黄溶液）> 碘酚（含 3%碘的液化苯酚）> 碘甘油（含 10%碘的甘油）> 稀碘酊（3%碘溶液）≈碘酊（6%碘溶液）≈ 碘伏（10%聚维酮碘溶液）> 双氧水（3%过氧化氢溶液）。这些结果提供了防腐剂对人体细胞相对毒性的有用估计，可与抗微生物活性进行比较，以在毒性最小的情况下最大化治疗益处。
