Open AccessLow concentrations of TNF ‐α promote osteogenic differentiation via activation of the ephrinB2‐EphB4 signalling pathway
Author(s) -
Wang Limei,
Zhang Jin,
Wang Cunwei,
Qi Yuping,
Du Mi,
Liu Wenhua,
Yang Chengzhe,
Yang Pishan
Publication year - 2017
Publication title -
cell proliferation
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.647
H-Index - 74
eISSN - 1365-2184
pISSN - 0960-7722
DOI - 10.1111/cpr.12311
Subject(s) - runx2 , tumor necrosis factor alpha , chemistry , blot , cell growth , microbiology and biotechnology , signal transduction , cell , in vitro , cellular differentiation , 3t3 cells , cell counting , biochemistry , biology , immunology , cell cycle , transfection , osteoblast , gene
Objectives Low concentrations of tumour necrosis factor‐alpha ( TNF ‐α) have been reported to promote osteogenic differentiation. In this study, a series of in vitro experiments was performed to investigate underlying molecular mechanisms involved. Materials and methods MC 3T3‐E1 murine preosteoblasts were treated with TNF ‐α at doses of 0, 0.1 or 1 ng/mL. The ephrinB2‐EphB4 signalling pathway was activated using ephrinB2‐fc, or inhibited using lentiviruses encoding si RNA s specifically targeting EphB4. Cell proliferation/survival was evaluated using the Cell Counting Kit‐8 ( CCK ‐8) assay, and expression levels of Runx2, BSP , ephrinB2 and EphB4 were determined using RT ‐ PCR and Western blotting. ALP activity in these cells was also determined, and mineral nodule formation was evaluated with alizarin red S staining. Results Low concentrations of TNF ‐α had no influence on cell proliferation/survival. However, expression levels of Runx2, BSP , ephrinB2 and EphB4, as well as ALP activity and mineral nodule formation, were significantly enhanced in MC 3T3‐E1 cells treated with low concentrations of TNF ‐α. Moreover, activation of the ephrinB2‐EphB4 signalling pathway by ephrinB2‐fc enhanced TNF ‐α‐induced osteogenic differentiation, while down‐regulation of EphB4 level reversed the positive effect of TNF ‐α. Conclusions Low concentrations of TNF ‐α promoted osteogenic differentiation via activation of the ephrinB2‐EphB4 signalling pathway.