Alpar B, Leyhausen G, Sapotnick A, Günay H, Geurtsen W
Department of Conservative Dentistry and Periodontology, Medical University Hannover, Germany.
Clin Oral Investig. 1998 Mar;2(1):40-6. doi: 10.1007/s007840050042.
Various in vivo and in vitro investigations have indicated that tobacco smoking as well as the use of smokeless tobacco products may be important risk factors for the development and severity of inflammatory periodontal disease. The purpose of this study was to determine the cytotoxicity of nicotine by means of human primary oral fibroblast cultures and a permanent cell line. The cytotoxicity of nicotine was evaluated by determination of cell growth, cell membrane integrity, protein content, and alterations of the cytoskeleton. Furthermore, recovery following nicotine exposure was assessed by vital staining (trypan blue). Dose-dependent toxic effects of nicotine were measured within a range of 0.48 mM to 62 mM. Growth of fibroblasts was decreased by nicotine concentrations higher than 7.8 mM. Additionally, the protein content was significantly decreased and cell membranes were damaged. Morphological alterations of microtubules and vimentin filaments were observed at concentrations higher than 3.9 mM. Nicotine-exposed cells revealed atypical shapes and vacuoles. The toxic effects of nicotine became irreversible in the range between 10.5 and 15.5 mM, whereas at lower concentrations cells recovered after the withdrawal of nicotine. Our results confirm clinical oberservations regarding the important role of nicotine as a risk factor in the etiology and progression of periodontal disease.
多项体内和体外研究表明，吸烟以及使用无烟烟草制品可能是炎症性牙周病发生和严重程度的重要危险因素。本研究的目的是通过人原代口腔成纤维细胞培养和一个永生细胞系来确定尼古丁的细胞毒性。通过测定细胞生长、细胞膜完整性、蛋白质含量和细胞骨架的改变来评估尼古丁的细胞毒性。此外，通过活细胞染色（台盼蓝）评估尼古丁暴露后的恢复情况。在0.48 mM至62 mM的范围内测定了尼古丁的剂量依赖性毒性作用。高于7.8 mM的尼古丁浓度会降低成纤维细胞的生长。此外，蛋白质含量显著降低，细胞膜受损。在高于3.9 mM的浓度下观察到微管和波形蛋白丝的形态改变。暴露于尼古丁的细胞呈现出非典型形状和空泡。尼古丁的毒性作用在10.5至15.5 mM之间变得不可逆，而在较低浓度下，尼古丁撤除后细胞会恢复。我们的结果证实了关于尼古丁作为牙周病病因和进展中的危险因素的重要作用的临床观察。
