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The induction of CXCR4 expression in human osteoblast‐like cells (MG63) by CoCr particles is regulated by the PLC‐DAG‐PKC pathway
Author(s) -
Drynda Andreas,
Ren Qiang,
Buchhorn Gottfried H.,
Lohmann Christoph H.
Publication year - 2017
Publication title -
journal of biomedical materials research part b: applied biomaterials
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.665
H-Index - 108
eISSN - 1552-4981
pISSN - 1552-4973
DOI - 10.1002/jbm.b.33770
Subject(s) - chelerythrine , protein kinase c , mapk/erk pathway , jurkat cells , osteolysis , signal transduction , microbiology and biotechnology , osteoblast , progenitor cell , osteoclast , kinase , staurosporine , chemistry , medicine , biology , immunology , receptor , stem cell , t cell , biochemistry , surgery , in vitro , immune system
Background Osteolysis which leads to aseptic loosening of implants is a fundamental problem in joint replacement surgery (arthroplasty) and the leading cause for implant failure and revision surgery. Metal (CoCr) particles separated from implants by wear cause osteolysis and the failure of orthopedic implants, but the molecular mechanism is not clear. The chemokine receptor CXCR4 has been shown to play a pivotal role in periprosthetic osteolysis. The aim of this study was to determine which signal transduction pathway (PLC‐DAG‐PKC or MAPK/ERK) induces CXCR4 expression in osteoblast‐like cells (MG63) cells. Methods MG63 and Jurkat cells were stimulated with different amounts of particles (10 7 , 10 6 , and 10 5 ) for different time periods (30 min to 24 h), in the presence and absence of specific inhibitors (chelerythrine for the PLC‐DAG‐PKC pathway and PD98059 for the MAPK/ERK pathway). The expression of CXCR4‐specific mRNA was determined by real‐time polymerase chain reaction (PCR), and the PKC activity was measured by Western Blot using an antibody specific for PKC‐related phosphorylation. Results Real‐time PCR data showed that CXCR4 mRNA expression in MG63 cells induced by CoCr particles was significantly diminished by the PKC‐specific inhibitor chelerythrine. This effect was not observed with the MAPK/ERK inhibitor PD98059. The involvement of PKC was also confirmed by an intensified phosphorylation pattern after stimulation with CoCr particles. In Jurkat cells, none of the inhibitors exhibited any effect. Conclusion The induction of CXCR4‐specific mRNA expression in MG63 cells after stimulation with CoCr particles is regulated by the PLC‐DAG‐PKC pathway and not by the MAPK/ERK pathway. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 105B: 2326–2332, 2017.