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Although the cariostatic effects of CO2 laser on the root surface have been shown, there is scarce information regarding its effects on root secondary caries. The objective of this research was to investigate the effect of the association of CO2 laser and a fluoride dentifrice on the inhibition of secondary caries on root surfaces adjacent to composite-resin or glass-ionomer-cement restorations. Dental blocks of human roots were divided into two groups: composite resin (CR) or glass ionomer cement (GIC). Subsequently, the blocks were divided into four subgroups (n  =  10): C, non-fluoride dentifrice; FD, fluoride dentifrice; L, CO2 laser with an energy density of 6.0 J cm−2  +  non-fluoride dentifrice; and L  +  FD, CO2 laser  +  fluoride dentifrice. The blocks were subjected to pH cycling to simulate a high cariogenic challenge. Dental demineralization around the restorations was quantified by microhardness analysis. The results were subjected to analysis of variance (ANOVA) and the Tukey–Kramer test (p  ⩽  0.05). As for mineral loss, it can be observed that all the groups that were treated with a fluoride dentifrice and laser, used alone or not, were statistically similar and superior to the RC–C group. It was concluded that CO2 laser irradiation and a fluoride dentifrice used alone or combined with each other are efficient surface treatments for preventing secondary root caries, regardless of the restorative material used. [ABSTRACT FROM AUTHOR]