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Abstract Objective To investigate the effect of osteocytes on the differentiation and formation of osteoclasts under the infection of Staphylococcus aureus.Methods The osteocytes were infected with Staphylococcus aureus for 4 hours.Staphylococcus aureus was inactivated.After changing the culture medium，the cells were cultured for another 72 hours，and then，the culture medium was collected and named as conditioned medium (CM) derived from osteocytes.After that，the osteoclast precursor cells (RAW264.7 cells) were induced with this conditioned medium (Induction group)，equal volume phosphate buffered saline (PBS，blank control group)，and receptor activator of nuclear factor κB ligand (RANKL，positive control group) for 5 days.In order to evaluate the effect of osteocytes on osteoclast differentiation，the number of osteoclasts was counted by tartrate resistant acid phosphatase (TRAP) staining，and the expression of osteoclast differentiation marker genes was detected by real timequantitative polymerase chain reaction (RT-qPCR).Results After 5 days of induction，TRAP staining showed that there was less osteoclast formation in the blank control group (PBS group)，but many osteoclasts were observed in the conditioned medium induction group (30% CM group) and the positive control group (RANKL group).After statistical analysis，the difference between the groups was statistically significant (P＜0.05)，and the number of osteoclasts formed in RANKL group was more than that in 30% CM group，more than that in PBS group.Moreover，RT-qPCR results showed that after 5 days induction，the expression of the marker genes of osteoclast differentiation in RANKL group and 30% CM group.Trap，calcitonin receptor，cathepsin K，MMP-9 were higher than that in PBS group.The difference between groups was statistically significant(P＜0.05).Conclusion Under the infection of Staphylococcus aureus,osteocytes can promote the differentiation and formation of osteoclasts by secreting some active molecules，and then participate in the pathological process of inflammatory bone destruction.
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Corresponding Authors:
Ren Lirong
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