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Wnt5a suppresses osteoblastic differentiation of human periodontal ligament stem cell‐like cells via Ror2/JNK signaling
Author(s) -
Hasegawa Daigaku,
Wada Naohisa,
Yoshida Shinichiro,
Mitarai Hiromi,
Arima Mai,
Tomokiyo Atsushi,
Hamano Sayuri,
Sugii Hideki,
Maeda Hidefumi
Publication year - 2018
Publication title -
journal of cellular physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.529
H-Index - 174
eISSN - 1097-4652
pISSN - 0021-9541
DOI - 10.1002/jcp.26086
Subject(s) - periodontal ligament stem cells , wnt signaling pathway , microbiology and biotechnology , gene knockdown , downregulation and upregulation , signal transduction , runx2 , chemistry , cellular differentiation , biology , osteoblast , alkaline phosphatase , biochemistry , gene , in vitro , enzyme
Wnt5a, a non‐canonical Wnt protein, is known to play important roles in several cell functions. However, little is known about the effects of Wnt5a on osteoblastic differentiation of periodontal ligament (PDL) cells. Here, we examined the effects of Wnt5a on osteoblastic differentiation and associated intracellular signaling in human PDL stem/progenitor cells (HPDLSCs). We found that Wnt5a suppressed expression of bone‐related genes ( ALP , BSP , and Osterix ) and alizarin red‐positive mineralized nodule formation in HPDLSCs under osteogenic conditions. Immunohistochemical analysis revealed that a Wnt5a‐related receptor, receptor tyrosine kinase‐like orphan receptor 2 (Ror2), was expressed in rat PDL tissue. Interestingly, knockdown of Ror2 by siRNA inhibited the Wnt5a‐induced downregulation of bone‐related gene expression in HPDLSCs. Moreover, Western blotting analysis showed that phosphorylation of the intracellular signaling molecule, c‐Jun N‐terminal kinase (JNK) was upregulated in HPDLSCs cultured in osteoblast induction medium with Wnt5a, but knockdown of Ror2 by siRNA downregulated the phosphorylation of JNK. We also examined the effects of JNK inhibition on Wnt5a‐induced suppression of osteoblastic differentiation of HPDLSCs. The JNK inhibitor, SP600125 inhibited the Wnt5a‐induced downregulation of bone‐related gene expression in HPDLSCs. Additionally, SP600125 inhibited the Wnt5a‐induced suppression of the alizarin red‐positive reaction in HPDLSCs. These results suggest that Wnt5a suppressed osteoblastic differentiation of HPDLSCs through Ror2/JNK signaling. Non‐canonical Wnt signaling, including Wnt5a/Ror2/JNK signaling, may function as a negative regulator of mineralization, preventing the development of non‐physiological mineralization in PDL tissue.