Background: Tooth whitening with peroxide is linked to a number of negative consequences. This study aimed to evaluate the effect of tooth bleaching and color safety using no-ozone cold atmospheric-pressure plasma (NCP) with distilled water (DW) and to assess the microhardness and mineral composition of the treated teeth. Methods: The enamel of 90 extracted human teeth were randomly assigned to three groups (n = 30): Group 1 (no treatment), Group 2 (NCP + DW), and Group 3 (only DW). The overall color change (ΔE) was observed at 10 min, 20 min, 30 min, 1 day, 7 days, 1 month, 3 months, and 6 months after treatment and quantified. Six cadavers treated with NCP and DW for 30 min were evaluated for color change using ShadeEye NCC and the VITA Easyshade. Furthermore, the microhardness and mineral composition of the teeth were evaluated using a Vickers hardness tester and an electro micro analyzer, respectively. All statistical data were analyzed at a significance level of 5%. Results: Group 1 showed no color change. The ΔE values of Group 2 were 17.67 and 5.62 times higher than those of Group 3 after 30 min and 6 months, respectively. Methylene blue solution revealed that the concentration of hydroxyl radicals was higher in Group 2. The average ΔE value using ShadeEye NCC for cadaveric teeth was 4.0. Evaluation via VITA Easyshade showed that the brightness was increased by 5 levels. No significant differences were observed among the groups in terms of the microhardness of the enamel and dentin or the Ca, P, Na, Cl, Zn, and Mg values in the enamel and dentin. Conclusions: Tooth bleaching using NCP with DW exhibited high efficacy without inducing damage or demineralization in the dental hard tissue.
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