Abstract Objectives Exploring the role of OPN N‐glycosylation in osteoblasts and osteoclasts. Methods Immunohistochemistry was used to detect the expression of OPN in mice with apical periodontitis. The asparagine at position 79 of the OPN protein was mutated to glutamine, and the above plasmids were transfected into osteoblasts and osteoclasts. The effect of OPN N‐glycosylation on proliferation of osteoblasts and osteoclasts was detected by CCK8 assays. Western blotting was used to detect the expression of OPN N‐glycosylation on osteoclasts and osteoblasts. Detection of N‐glycosylation of OPN activated the NF‐κB signaling pathway to regulate osteoblasts and osteoclasts. Results OPN increased the expression in a mice model of apical periodontitis. The expression curve of OPN resembled a reverse V shape. The OPN N‐glycosylation site was identified as 79 by MS. N‐glycosylation of OPN promoted the proliferation of osteoclasts. But the N79 glycosylation site of mutant OPN could not increase the proliferation of osteoblasts. OPN N‐glycosylation modulated the expression of osteoclast‐ and osteoblast‐associated factors through the NF‐κB signaling pathway. N‐glycosylation of OPN promoted nuclear translocation of NF‐κB in osteoclasts and osteoblasts. Conclusions The N‐glycosylation site of OPN is 79. N‐glycosylation of OPN played an important role in the biological function of OPN protein.