Staphylococci planktonic and biofilm environments differentially affect macrophage immune activation and osteoclastogenic differentiation

Abstract Implant-related bone infections are a major complication in orthopedic surgery that lead to inflammation and bone destruction. Bacterial biofilm formation on the implant is discussed to polarize the immune response towards tolerance and to facilitate bacterial persistence. In addition to their role in the early immune response, macrophages are osteoclast precursor cells. Therefore, macrophages can link inflammation and RANKL-mediated osteoclastogenic bone destruction. We investigated the influence of Staphylococcus aureus (SA) and epidermidis (SE) biofilm formation on immune function and osteoclastogenesis using RAW264.7 cells and conditioned media (CM) of planktonic and biofilm cultures in the presence and absence of the osteoclastogenic transcription factor RANKL. Analysis of immune cell activation, metabolic activity and osteoclast formation revealed that a planktonic environment causes a pro-inflammatory response. This was also partially induced by biofilm CM. Simultaneous stimulation with CM and RANKL suppressed osteoclast formation in favor of a long-term immune activation. While the early macrophage response towards CM was dominated by glycolysis, the CM and RANKL approach shifted metabolism towards increased mitochondrial biomass and activity. This was most evident in biofilm CM. We further showed that planktonic CM effects are mediated through activation of TLR signaling and induction of IFN-β production. In biofilm CM, high lactate levels seem to significantly contribute to the modulation of macrophages. Our results can contribute to find targets for therapeutic intervention that restore an effective pro-inflammatory immune response, which could help to control implant-related bone infections. Importance The number of endoprosthetic and bone reconstructive surgeries is rising and accordingly incidences of implant-related bone infections increase. Biofilm formation on the implant surface protects bacteria against most antibiotics and the host’s immune response. Thus, novel therapeutic strategies for prevention or treatment of biofilm formation are needed. Furthermore, bone infections are associated with osteoclast-mediated bone destruction, which promotes infection progression and ultimately leads to implant loosening. We investigated the effect of staphylococci-mediated biofilm formation on macrophage immune activation and osteoclastogenic differentiation, respectively. In planktonic and biofilm environments, immune cell activation dominated over osteoclastogenesis. The effects of planktonic environments were mainly mediated through activation of TLR signaling and resulted in a pronounced pro-inflammatory and IFN-β macrophage immune response. In biofilm environments, high lactate levels impaired osteoclast formation but also prevented an effective long-term immune activation. Therefore, targeting biofilm-derived metabolites but also IFN-β signaling might be promising novel treatment strategies.