Abstract: Objective: To investigate the inhibitory molecules repressing osteogenic differentiation of PDLSCs in inflammatory microenvironment.Methods:The osteogenic ability of H-PDLSCs and P-PDLSCs was investigated. The expressions of GSK3β/p-GSK3β and β-catenin in H-PDLSCs and P-PDLSCs were examined by Western blot.Luciferase assay was performedto studyβ-catenin/TCF transcriptional activity,using the Dual Luciferase Assay System kit. RNAinterference technique was used to suppressβ-catenin levels in PDLSCs.Human recombinant Wnt3aand lithium chloride (LiCl) was applied during osteogenesis and used Alizarin red stainingto investigate the osteogenic differentiation of H-PDLSCs.Results:PDLSCs from periodontitis patients had impaired differentiation capacity.More strikingly enhanced p-GSK3β and β-catenin were found in P-PDLSCs than H-PDLSCs.Inhibition of β-cateninpreserved the osteogenesis of TNF-α-stimulated PDLSCs. Alizarin red staining showed that the osteogenic differentiation of PDLSCs was decreased significantly after LiCl or Wnt3a stimulation, which had the same effect as TNF-α.Conclusions:GSK3β was required forTNF-α-mediated inhibition of osteogenicdifferentiation in PDLSCs, which may provide a novel potential approach tobone regeneration in inflammatory microenvironments.