Objective: An amelogenin-derived peptide has been shown to promote remineralization of demineralized enamel in an in vitro model of initial caries induced by pH cycling. The present study examines whether the peptide exerts similar effects within the complex oral environment in vivo. Design: Specific pathogen-free Sprague-Dawley rats (n =36) were infected with Streptococcus mutans, given ad libitum access to Diet 2000 and drinking water supplemented with sucrose (10%, w/v), and then randomly divided into three groups treated with 25 mu M peptide solution, 1 g/L NaF or deionized water. Molar teeth were swabbed twice daily with the respective solutions for 24 days. Then animals were killed, their jaws were removed and caries lesions were analyzed using the quantitative light-induced fluorescence-digital (QLF-D) technique to measure changes in mineral content. To verify QLF-D results, caries were scored for lesion depth and size using the Keyes method, and analyzed using polarized light microscopy (PLM). Results: Mineral gain was significantly higher in teeth treated with peptide or NaF than in teeth treated with water (p < 0.05), based on the QLF-D results (Delta F and Delta Q). Incidence of smooth-surface and sulcal caries based on Keyes scores was similar in rats treated with peptide or NaF, and significantly lower in these groups than in rats treated with water (p < 0.05). Lesions on teeth treated with peptide or NaF were shallower, based on PLM. No significant differences were observed between molar enamel caries treated with peptide or NaF. Conclusions: This amelogenin-derived peptide can promote remineralization in a rat caries model, indicating strong potential for clinical use. (C) 2016 Elsevier Ltd. All rights reserved.