Enamel matrix derivative-induced cementoblast differentiation via Wnt/β-catenin signaling in human periodontal ligament cells

Abstract

Purpose: Cementum is a specialized calcified tissue produced by cementoblasts, which play a key role in periodontal regeneration. Enamel matrix derivative (EMD) is an agent currently employed for treatment of periodontal disease, although its molecular action mechanism remains unclear. The present study aimed to investigate the effects of EMD on cementoblast differentiation and the involvement of Wnt/β-catenin signaling in human periodontal ligament (hPDL) cells.

Methods: Primary hPDL cells were treated with EMD, and the expression of cementum-related markers was evaluated. The role of Wnt/β-catenin signaling was assessed using IWP-2, a Wnt inhibitor. A luciferase reporter assay was conducted to identify regulatory elements within the cementum protein 1 (CEMP1) promoter.

Results: EMD upregulated the expression of CEMP1 and cementum attachment protein. This effect was attenuated by IWP-2, indicating Wnt/β-catenin involvement. A T-cell factor binding site was identified within the CEMP1 promoter. EMD increased the expression of osterix and runt-related transcription factor 2 (Runx2) slightly, but decreased the levels of osteocalcin and osteopontin. No mineralization enhancement was observed based on alkaline phosphatase activity and Alizarin Red S stainingConclusion: EMD promotes early-stage cementoblast differentiation via Wnt/β-catenin signaling but has limited effects on late-stage maturation, offering new insights into the regenerative potential of EMD for periodontal therapy.

Keywords: Wnt signaling pathway; cementoblast; enamel matrix derivative; periodontal regeneration.