Occlusion of dentine tubules may reduce or eliminate dentin hypersensitivity. Here, micro-nano bioactive glass spheres (MNBGs) with various sizes and sodium alginate-phosphate buffer solution were used to prepare bioactive glass pastes (MNBGP) for dentin desensitization. Through mineralization on the surface of dentin samples in vitro, the bonding ability of different pastes with dentin, as well as the ability to induce dentin remineralization and occlude dentin tubules in vitro, was investigated. The results indicated that the MNBGP could be tightly combined with the dentin interface, and the MNBGs with smaller particle sizes were more uniformly distributed on the surface of the demineralized dentin slices. MNBGP could well mineralize in artificial saliva (AS) to form apatite (HA), induce dentin remineralization, and occlude dentin tubules, which resulted in thickening HA layer, formed on the surface of demineralized dentin slices, increased with the prolonged mineralization time, up to 5-10 μm after 28 d dentin remineralization. The size of bioactive glass microspheres had a certain impact on its ability to induce remineralization. When the particle size was equal to the diameter of the exposed dentin tubules, it could better occlude the dentin tubules. All data indicate that MNBGP has potential application for dentin hypersensitivity treatment.