Canonical Wnt/β-catenin signaling has positive effects on osteogenesis, but can have negative effects on cementogenesis.

Abstract

To date, therapeutic approaches for cementum regeneration are limited and outcomes remain unpredictable. A significant barrier to improve therapies for cementum regeneration is that the cementocyte and its intracellular signal transduction mechanisms remain poorly understood. This study aims to elucidate the regulatory mechanism of Wnt pathway in cementogenesis. The effects of canonical Wnt signaling were compared in vitro using immortalized murine cementocyte cell line IDG-CM6 and osteocyte cell line IDG-SW3 by quantitative real-time polymerase chain reaction, Western blot, confocal microscopy, alkaline phosphatase (ALP) assay, and Alizarin red S staining. In vivo, histological changes of cementum and bone formation were examined in transgenic mice in which constitutive activation of β-catenin is driven by Dmp1 promoter. Expression of components of the Wnt/β-catenin pathway were much greater in the IDG-SW3 cells compared with the IDG-CM6 cells resulting in much lower expression of Sost/sclerostin in the IDG-SW3 cells. In the IDG-CM6 cells, low dose Wnt3a (20ng/ml) had a modest effect while high dose (200ng/ml) inhibited runt-related transcription factor 2, osterix, ALP, and osteopontin in contrast to the IDG-SW3 cells where high dose Wnt3a dramatically increased mRNA expression of these same markers. However, high Wnt3a significantly increased mRNA for components of Wnt/β-catenin signaling pathway in both IDG-CM6 and IDG-SW3 cells. In vivo, constitutive activation of β-catenin in the Dmp1-lineage cells in mice leads to bone hyperplasia and cementum hypoplasia. These findings indicate that Wnt signaling has distinct and different effects on the regulation of long bone as compared with cementum.