Kleinsasser N H, Weissacher H, Wallner B C, Kastenbauer E R, Harréus U A
Klinisch experimentelle Onkologie der Klinik und Poliklinik für Hals-, Nasen- und Ohrenkranke der Ludwig-Maximilians-Universität München.
Laryngorhinootologie. 2001 Apr;80(4):187-90. doi: 10.1055/s-2001-13760.
Fluorides are widely used in dental health products and drinking water, due to their beneficial effects in caries-prophylaxis and -treatment. Nevertheless, irritation of the gingiva and oropharyngeal mucosa as well as in gastric mucosa is observed since neither local nor systemic application is restricted to the teeth. These effects may partly be attributed to a known cytotoxicity of fluorides. Whether fluorides also have genotoxic effects on human mucosa or lymphocytes as a possible factor in tumor initiation was investigated in this study.
Human oropharyngeal epithelial cells and peripheral lymphocytes were incubated after single cell preparation with the aminefluoride Olaflur at concentrations of 2 ppm, 21 ppm, 35 ppm, 71 ppm and 213 ppm. The extent of cytotoxicity was investigated using the trypan blue exclusion test. Following incubation, electrophoresis for migration of DNA fragments, fluorescence staining and digital image analysis according to a standard protocol of the single cell microgel electrophoresis assay (Comet assay) followed. DNA damage was characterized using the Olive Tail Moment (OTM).
For fluoride concentrations of 2 ppm to 35 ppm, non vital cells of less than 10% could be shown. After incubation with 71 ppm and 213 ppm Olaflur, there were 15% and 43% of damaged cells, respectively. Weak genotoxic effects on mucosal cells as well as on lymphocytes could be demonstrated at all concentrations tested. In fluoride concentrations of 213 ppm genotoxicity increased to max. OTM-levels of 23.
Beside the cytotoxic effect of fluorides, also a minor genotoxic impact on human mucosa and on peripheral lymphocytes could be demonstrated using the Comet assay. Further investigations are warranted to examine fluorides in a model allowing for repeated or long term incubations on structurally intact human mucosa in vitro. Such a model will help to distinguish between DNA damage that may be repaired successfully and other impairments that may show an additive character in repetitive or chronic exposure in vivo.
由于氟化物在预防和治疗龋齿方面具有有益作用,因此广泛应用于牙科保健产品和饮用水中。然而,由于局部或全身应用并不局限于牙齿,因此会观察到牙龈、口咽黏膜以及胃黏膜受到刺激。这些影响可能部分归因于氟化物已知的细胞毒性。本研究调查了氟化物是否也对人类黏膜或淋巴细胞具有基因毒性作用,这可能是肿瘤发生的一个因素。
将人源口咽上皮细胞和外周淋巴细胞单细胞制备后,分别用浓度为2 ppm、21 ppm、35 ppm、71 ppm和213 ppm的胺氟化物奥拉氟进行孵育。使用台盼蓝排斥试验研究细胞毒性程度。孵育后,按照单细胞微凝胶电泳试验(彗星试验)的标准方案进行DNA片段迁移电泳、荧光染色和数字图像分析。使用橄榄尾矩(OTM)对DNA损伤进行表征。
对于2 ppm至35 ppm的氟化物浓度,显示非存活细胞少于10%。用71 ppm和213 ppm奥拉氟孵育后,受损细胞分别为15%和43%。在所有测试浓度下,均可证明对黏膜细胞和淋巴细胞有微弱的基因毒性作用。在213 ppm的氟化物浓度下,基因毒性增加至最大OTM水平23。
除了氟化物的细胞毒性作用外,使用彗星试验还可证明其对人类黏膜和外周淋巴细胞有轻微的基因毒性影响。有必要进行进一步研究,以在一个允许对结构完整的人源黏膜进行重复或长期体外孵育的模型中检测氟化物。这样一个模型将有助于区分可能成功修复的DNA损伤和在体内重复或慢性暴露中可能表现出累加特性的其他损伤。