Original Research Report
The induction of CXCR4 expression in human osteoblast-like cells (MG63) by CoCr particles is regulated by the PLC-DAG-PKC pathway
Andreas Drynda,
Qiang Ren,
Gottfried H. Buchhorn,
Christoph H. Lohmann,
Andreas Drynda
Department of Orthopaedic Surgery, Otto-von-Guericke University, Magdeburg, Germany
Search for more papers by this authorQiang Ren
Department of Orthopaedic Surgery, Otto-von-Guericke University, Magdeburg, Germany
Search for more papers by this authorGottfried H. Buchhorn
Department of Orthopaedics, University Medical Center, Göttingen, Germany
Search for more papers by this authorCorresponding Author
Christoph H. Lohmann
Department of Orthopaedic Surgery, Otto-von-Guericke University, Magdeburg, Germany
Correspondence to: C. H. Lohmann; e-mail: [email protected]Search for more papers by this authorAndreas Drynda,
Qiang Ren,
Gottfried H. Buchhorn,
Christoph H. Lohmann,
Andreas Drynda
Department of Orthopaedic Surgery, Otto-von-Guericke University, Magdeburg, Germany
Search for more papers by this authorQiang Ren
Department of Orthopaedic Surgery, Otto-von-Guericke University, Magdeburg, Germany
Search for more papers by this authorGottfried H. Buchhorn
Department of Orthopaedics, University Medical Center, Göttingen, Germany
Search for more papers by this authorCorresponding Author
Christoph H. Lohmann
Department of Orthopaedic Surgery, Otto-von-Guericke University, Magdeburg, Germany
Correspondence to: C. H. Lohmann; e-mail: [email protected]Search for more papers by this authorAbstract
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 (107, 106, and 105) 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.
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