Hypothesis: Dental bleaching with H2O2 is a common daily practice in dentistry to correct discoloration of anterior teeth. The aim of this study has been to determine whether this treatment of human teeth affects growth, differentiation and activity of osteoclast-like cells, as well as the putative modulatory action of osteostatin and fibroblast growth factor 2 (FGF-2). Experiments: Previously to the in vitro assays, structural, physicalchemical and morphological features of teeth after bleaching were studied. Osteoclast-like cells were cultured on human dentin disks, pretreated or not with 38% H2O2 bleaching gel, in the presence or absence of osteostatin (100 nM) or FGF-2 (1 ng/ml). Cell proliferation and viability, intracellular content of reactive oxygen species (ROS), proinflammatory cytokine (IL-6 and TNFα) secretion and resorption activity were evaluated. Findings: Bleaching treatment failed to affect either the structural or the chemical features of both enamel and dentin, except for slight morphological changes, increased porosity in the most superficial parts (enamel), and a moderate increase in the wettability degree. In this scenario, bleaching produced an increased osteoclast-like cell proliferation but decreased cell viability and cytokine secretion, while it augmented resorption activity on dentin. The presence of either osteostatin or FGF-2 reduced the osteoclast-like cell proliferation induced by bleaching. FGF-2 enhanced ROS content, whereas osteostatin decreased ROS but increased TNFα secretion. The bleaching effect on resorption activity was increased by osteostatin, but this effect was less evident with FGF-2. Conclusions: These findings further confirm the deleterious effects of tooth bleaching by affecting osteoclast growth and function as well as different modulatory actions of osteostatin and FGF-2. We sincerely appreciate reviewers' remarks to improve the article and we have modified our manuscript following their recommendations. We have included additional physical-chemical and structural characterization to determine the effects with the bleaching. Moreover, the Conclusion has been improved in the revised manuscript according to your technical notes. The modifications are highlighted in the manuscript and detailed responses to the reviewers' comments are included bellow.
ProfWe hope that the revised version of our manuscript is now appropriate for publication in Journal of Colloid and Interface Science.Thank you for your kind consideration of our work.
Sincerely,Prof. María Vallet-Regí.
1: Cover LetterResponse to the Reviewers:Reviewers' comments:Reviewer #2: The ms reports on an investigation of the effects of teeth bleaching by H2O2 on osteoclast activities and their modulation by osteostatin and fibroblast growth factor 2. The ms provides a nice brief biological investigation of a well-specified issues, but essentially no surface chemical information. Therefore, the authors must address the following issues before this ms can be further considered for publication:1) Surfa...