Polymorphonuclear Neutrophils in Osteomyelitis: Link to Osteoclast Generation and Bone Resorption

Abstract

Chronic and persistent inflammatory processes in bones may lead to severe erosions with consequent functional impairment sometimes requiring amputation of the limb. To explore the relationship between inflammation and bone erosion, biopsies of patients with osteomyelitis due to arterial occlusive disease or to diabetes mellitus were examined (n=31). Histologically, inflammation and bone erosion were confirmed. In the eroded bones the number of osteoclasts correlated with the abundance of infiltrated polymorphonuclear neutrophils (PMN), which were highly activated as shown by expression of MHC class II. For functional characterisation of the infiltrating PMN, patients with implant-associated osteomyelitis, a condition associated with persistent bacterial infection and bone destruction, were recruited. The cells were recovered from infected sites and examined ex vivo. These PMN expressed MHC class II and produced interleukin (IL)-8, a further indication of PMN activation. To assess a possible link between infiltrating PMN and bone erosion, we tested the effect of IL-8 on osteoclast generation in vitro. CD14+ monocytes derived from the peripheral blood of healthy individuals were cultivated with monocyte colony stimulating factor (M-CSF) and IL-8. Within 3 days, a translocation of the transcription factor NFATcl into the nucleus was seen, and by 10 to 20 days multinucleated cells with typical osteoclast morphology appeared that expressed tartrate-resistant acid phosphatase (TRAP) and cathepsin K. Moreover, the cells were able to resorb bone, proving that IL-8 was able to induce the differentiation of monocytes to osteoclasts. Because IL-8 is a major cytokine produced by activated PMN, we propose that in the course of persistent infection infiltrating PMN contribute to induction of osteoclast formation, thus providing a link between inflammation and bone erosion.