Enterococcus faecalis attenuates osteogenesis through activation of p38 and ERK1/2 pathways in MC3T3-E1 cells
S. Wang
Department of Endodontics, Comprehensive Dental Care, Faculty of Dentistry, The University of Hong Kong, Hong Kong SAR, China
S. Wang and Z. Deng contributed equally to this work.
Search for more papers by this authorZ. Deng
Department of Endodontics, Comprehensive Dental Care, Faculty of Dentistry, The University of Hong Kong, Hong Kong SAR, China
Shenzhen ENT Institute, Shenzhen, China
S. Wang and Z. Deng contributed equally to this work.
Search for more papers by this authorX. Ye
Department of Endodontics, Comprehensive Dental Care, Faculty of Dentistry, The University of Hong Kong, Hong Kong SAR, China
School of Dentistry, Shandong University, Jinan, China
Search for more papers by this authorX. Geng
Department of Stomatology, Shenzhen Longgang Center Hospital, ENT Hospital, Shenzhen, China
Search for more papers by this authorCorresponding Author
C. Zhang
Department of Endodontics, Comprehensive Dental Care, Faculty of Dentistry, The University of Hong Kong, Hong Kong SAR, China
Correspondence: Chengfei Zhang, Comprehensive Dental Care, Faculty of Dentistry, University of Hong Kong, 34 Hospital Road, Sai Ying Pun, Hong Kong SAR, China (Tel.: +852 2859 0287; Fax: +852 2559 9013; e-mail: [email protected]).Search for more papers by this authorS. Wang
Department of Endodontics, Comprehensive Dental Care, Faculty of Dentistry, The University of Hong Kong, Hong Kong SAR, China
S. Wang and Z. Deng contributed equally to this work.
Search for more papers by this authorZ. Deng
Department of Endodontics, Comprehensive Dental Care, Faculty of Dentistry, The University of Hong Kong, Hong Kong SAR, China
Shenzhen ENT Institute, Shenzhen, China
S. Wang and Z. Deng contributed equally to this work.
Search for more papers by this authorX. Ye
Department of Endodontics, Comprehensive Dental Care, Faculty of Dentistry, The University of Hong Kong, Hong Kong SAR, China
School of Dentistry, Shandong University, Jinan, China
Search for more papers by this authorX. Geng
Department of Stomatology, Shenzhen Longgang Center Hospital, ENT Hospital, Shenzhen, China
Search for more papers by this authorCorresponding Author
C. Zhang
Department of Endodontics, Comprehensive Dental Care, Faculty of Dentistry, The University of Hong Kong, Hong Kong SAR, China
Correspondence: Chengfei Zhang, Comprehensive Dental Care, Faculty of Dentistry, University of Hong Kong, 34 Hospital Road, Sai Ying Pun, Hong Kong SAR, China (Tel.: +852 2859 0287; Fax: +852 2559 9013; e-mail: [email protected]).Search for more papers by this authorAbstract
Aim
To explore the role of Enterococcus faecalis in the proliferation, apoptosis and differentiation of osteoblasts.
Methodology
Pre-osteoblastic MC3T3-E1 cells were treated with heat-killed E. faecalis ATCC 29212 and clinical E. faecalis P25RC strains, respectively. Cell proliferation, mineralized calcium deposition, alkaline phosphatase (ALP) activity and apoptosis were assessed at various time-points. The expression levels of osteogenic-related genes including ALP, osteocalcin (OC), runt-related protein 2 (Runx2) and collagen type 1 (COL1) were also analysed throughout the duration of the experiment. Additionally, the involvement of mitogen-activated protein kinases (MAPKs) signalling pathways was analysed by Western blotting. In the presence of culture supernatant from E. faecalis-treated murine macrophages, apoptosis of MC3T3-E1 cells was detected with flow cytometry. Data were analysed using analysis of variance (anova), and P < 0.05 was considered significantly different.
Results
E. faecalis significantly inhibited proliferation (P < 0.05) and also significantly induced apoptosis of MC3T3-E1 cells (P < 0.05), whilst differentiation seemed to be unaffected after 7 days of E. faecalis treatment. However, osteogenic differentiation was significantly inhibited with 21-day E. faecalis treatment (P < 0.05). The p38 and ERK1/2 phosphorylation pathways associated with mineral deposition and apoptosis were significantly activated in MC3T3-E1 cells. The culture supernatants from E. faecalis-treated macrophages induced osteoblast apoptosis.
Conclusions
E. faecalis exerted an inhibitory effect on osteogenesis in pre-osteoblastic MC3T3-E1 cells via phosphorylation of p38 and ERK1/2.
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