Smokeless Tobacco And Lipid Peroxidation Of Human Oral Keratinocytes

Normal human oral keratinocytes (NHOK) were incubated with STE (0 to 300^g/ml) for 24h at 37°C. Keratinocytes were also preincubated with vitamin C (75^M), vitamin E (75^M), a combination pf vitamins C and E (75^M each), and grape seed extract (GSPE) (100^g/ml) for 4h at 37°C. These optimal concentrations for vitamins C and E were selected based on our earlier experiments. Production of thiobarbituric acid reactive substances (TBARS), a commonly used technique that has been related to lipid peroxidation and tissue damage, was determined according to the method of Buege and Aust30 and as described previously.40 The production of TBARS was determined with NOHK cells following STE incubation. Concentration-dependent increases of 3.5-, 5.7-, and 7.6-fold in lipid peroxidation occurred at 100, 200, and 300^g/ml of STE, respectively, for 24-h posttreatment relative to control values. Preincubation of the cells with vitamin C, vitamin E, a combination of vitamins C plus E, and GSPE for 4 h decreased STE (300^g/ml)-induced lipid peroxidation by 22, 34, 43, and 54%, respectively, as compared with the corresponding STE-treated cells.2840

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