Calcium boosters have been used as a supplement for fluoride toothpastes to repair the dental tissues and reduce dentin permeability. This in vitro study aimed to characterize the regenerative and protective effects of the treatment of dental tissues with a fluoride–silicon-rich toothpaste associated with a calcium booster. Bovine enamel and dentin blocks (n= 5) were obtained (4 × 4 × 6 mm). A fluoride–silicon-rich toothpaste and a calcium booster were used to brush the enamel and dentin both immediately and five days afterwards. The outcomes were then compared to those of the untreated control group. After that, the specimens were cross-sectioned. SEM was used to evaluate the micromorphology of the surface and cross-section. Energy-dispersive X-ray spectroscopy (EDS) was used to determine the elemental analyses (weight%). After treatment for 5 days with a booster/silicon-rich toothpaste, EDS analysis demonstrated that it induced a significant mineral change. It was also able to form a protective silicon-enriched mineral layer on both enamel and dentin surfaces. It was demonstrated in vitro that a fluoride–silicon-rich toothpaste associated with a calcium booster regenerates the dental tissues, remineralizing the enamel structure and occluding the dentin tubules.