In situ and in vitro effects of bleaching with carbamide peroxide on human enamel

This study evaluated in vitro and in situ the potential adverse effects of 10% carbamide peroxide on human enamel using microhardness, calcium loss and surface morphology analysis. Twenty-four enamel slices (4 mm2) were obtained from recently extracted premolars. The specimens were polished under water-cooling down to 1,200-grade sandpaper. After initial microhardness readings (100 g), the specimens were randomly divided into two groups for in situ and in vitro conditions. The specimens were covered with 10% carbamide peroxide for eight hours. After removing the bleaching gel, the in vitro specimens were stored in deionized water and the in situ specimens, included in an intra-oral appliance, were placed in the oral cavity of four volunteers. These cycling sequences took place for 14 days. Upon conclusion of the bleaching treatment, new microhardness readings were performed on all specimens. Calcium dosage was assessed from the bleaching gel collected after initial exposure on day one, then from gel collected between days two and seven and gel collected between day eight and 14 using an atomic absorption spectrophotometer. Surface morphology was observed from two non-treated control specimens and two specimens of each experimental bleached group under SEM evaluation. Statistical analysis (ANOVA and Tukey tests) disclosed that specimens bleached in situ showed similar microhardness to unbleached specimens and had statistically higher (p < 0.01) hardness than in vitro bleached specimens. The loss of calcium in the in vitro situation at 14 days was 2.5 times higher than the in situ condition. SEM micrographs demonstrated that surface alterations were more pronounced in the in vitro condition. The adverse effects of carbamide peroxide on enamel were evident in specimens bleached in vitro but were not seen in situ. The presence of saliva could prevent the demineralizing effect of bleaching gel in situ.