Cigarette smoking is a risk factor for periodontal disease. The effect of cigarette smoke and nicotine, a major component of tobacco, on the morphological changes of human periodontal ligament fibroblasts (HPLF) was investigated.When suspended HPLF were incubated with cigarette smoke and nicotine solution in a 5% CO
2 atmosphere at 37°C, the cells did not neither adhered to the glass nor multiplied.When confluent HPLF were incubated with cigarette smoke and nicotine, shrinking of the cells, with the formation of intracellular spaces, and cellular lysis were observed. The changes depended on the nicotine concentrations and incubation periods. The expression of nicotine-induced cytokines was examined with a RayBio
TM human Cytokine Kit using a chemiluminescence method. IL-6 and IL-8, both inflammatory cytokines, were expressed in the HPLF at a low concentration of nicotine. Cell death was observed in the presence of nicotine, as detected by TUNEL. Apoptosis appeared when the cells were incubated with a high concentration of nicotine. The generation of free radicals in the cigarette smoke and nicotine solutions was measured by the spin trapping technique using an electron spin resonance spectrometer(ESR) to identify superoxide anions and hydroxyl radicals. When the HPLF were incubated with hexamethonium, an inhibitor of nicotine receptors, the cells showed altered morphology as well as nicotine-induced morphological changes.
These results suggest that the HPLF were destroyed via a direct local action through the generation of free radicals induced by nicotine.
Nihon Shishubyo Gakkai Kaishi (J Jpn Soc Periodontol) 50(4) : 238-249, 2008.
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