We investigated the remineralization effects of Nanoseal (NS) dentin desensitizer on demineralized root dentin. Baseline lesion specimens prepared from bovine root dentin were immersed in artificial saliva (AS) or deionized water (DW) after treatment with NS or fluoride-free Nanoseal (NS(−)). Treatment and control groups comprised: 1, AS; 2, NS/AS; 3, NS(−)/AS; 4,NS/DW; 5, NS(−)/DW; and 6, baseline demineralization. Integrated mineral loss (IML) and lesion depth (LD) were determined by transverse microradiography. Fluoride concentrations in the immersion solutions were measured. AS, NS/AS and NS(−)/AS showed higher mineral volume % at the surface and lesion body than did other groups. NS/AS showed significantly lower IML than did AS. There was no significant difference in IML between NS/AS and NS(−)/AS. The highest concentration of fluoride was in the NS/AS immersion solution. The findings suggest Nanoseal facilitated remineralization of demineralized root dentin, and fluoride and other ions included may have contributed to this effect.