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Clin Oral Investig. 2015 Apr;19(3):673-80. doi: 10.1007/s00784-014-1285-3. Epub 2014 Jul 19.

Color alteration, hydrogen peroxide diffusion, and cytotoxicity caused by in-office bleaching protocols.

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  • 1Department of Restorative Dentistry, Araçatuba School of Dentistry, Univ. Estadual Paulista (UNESP), Rua José Bonifácio, 11930, Araçatuba, SP, 14801-903, Brazil.



This study evaluated the color alteration, cytotoxicity, and hydrogen peroxide (HP) diffusion associated with different in-office bleaching protocols.


Bovine enamel/dentin disks were subjected to three bleaching sessions with 35 % HP (three 15-min applications), 35 % HP (one 45-min application), or 20 % HP (one 45-min application). The control group was not bleached. Before bleaching, the disks were adapted to artificial pulp chambers positioned in compartments containing 1 ml of acetate buffer or medium, so that the dentin remained in contact with these substances. Immediately after bleaching, the HP that diffused through the disks was stabilized by acetate buffer and was quantified (two-way repeated measures ANOVA/Fisher's protected least significant difference (PLSD) test; α = 5 %). Cells of mouse dental papilla cell-23 (MDPC-23) were incubated in this culture media for 1 h, followed by analysis of cellular metabolism (methyl tetrazolium assay) (one-way ANOVA/Tukey test; α = 5 %) and morphology (scanning electron microscopy). The specimen color alteration (ΔE) was analyzed by reflection spectrophotometry (two-way repeated measures ANOVA/Fisher's PLSD test; α = 5 %).


All protocols showed equal effectiveness at the end of the treatment. HP diffusion was significantly higher in the groups bleached with 35 % HP. Reapplication of 35 % HP resulted in increased diffusion only in the first session; however, the decrease in cell metabolism was similar for all studied protocols.


Despite greater peroxide diffusion in the groups treated with 35 % HP, all protocols showed the same effectiveness and were cytotoxic to MDPC-23 cells.


Bleaching protocols using high HP concentrations should be avoided because they exert aggressive actions on odontoblast-like cells.

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