Extracellular vesicles regulate the human osteoclastogenesis: divergent roles in discrete inflammatory arthropathies.

Abstract

Extracellular vesicles (EVs) are subcellular signalosomes. Although characteristic EV production is associated with numerous physiological and pathological conditions, the effect of blood-derived EVs on bone homeostasis is unknown. Herein we evaluated the role of circulating EVs on human osteoclastogenesis. Blood samples from healthy volunteers, rheumatoid arthritis (RA) and psoriatic arthritis (PsA) patients were collected. Size-based EV sub-fractions were isolated by gravity-driven filtration and differential centrifugation. To investigate the properties of EV samples, resistive pulse sensing technique, transmission electron microscopy, flow cytometry and western blot were performed. CD14+ monocytes were separated from PBMCs, and stimulated with recombinant human M-CSF, RANKL and blood-derived EV sub-fractions. After 7days, the cells were fixed and stained for tartrate-resistant acid phosphatase and counted. EVs isolated by size-based sub-fractions were characterized as either microvesicles or exosomes (EXO). Healthy (n=11) and RA-derived (n=12) EXOs profoundly inhibited osteoclast differentiation (70%, p<0.01; 65%, p<0.01, respectively). In contrast, PsA-derived (n=10) EXOs had a stimulatory effect (75%, p<0.05). In cross-treatment experiments where EXOs and CD14+ cells were interchanged between the three groups, only healthy (n=5) and RA (n=5)-derived EXOs inhibited (p<0.01, respectively) the generation of osteoclasts in all groups, whereas PsA (n=7)-derived EXOs were unable to mediate this effect. Our data suggest that blood-derived EXOs are novel regulators of the human osteoclastogenesis and may offer discrete effector function in distinct inflammatory arthropathies.