Amelogenin-derived peptide-5 suppresses the receptor activator of nuclear factor-κB ligand-induced osteoclast differentiation via adenosine monophosphate-activated protein kinase activation
DOI:
https://doi.org/10.1590/1678-7765-2026-0347Keywords:
Amelogenin-derived peptide-5, Odontoclasts, Osteoclast, Osteoclastogenesis, Root resorptionAbstract
Pathological root resorption is mediated by odontoclasts, which share functional characteristics with osteoclasts. However, the effect of amelogenin-derived peptide-5 (ADP-5) on osteoclast differentiation remains unclear. Objective This study aimed to determine whether ADP-5 inhibits the receptor activator of nuclear factor-κB ligand (RANKL)-induced osteoclast differentiation and to explain the involvement of adenosine monophosphate-activated protein kinase (AMPK). Methodology Bone marrow-derived macrophages were isolated from six-week-old mice from the Institute of Cancer Research. Cell viability was assessed. Osteoclast differentiation was induced with RANKL (100 ng/mL) and macrophage colony-stimulating factor (30 ng/mL) in the presence of ADP-5 (0–20 μM). Multinucleated cells were quantified by tartrate-resistant acid phosphatase (TRAP) staining. Osteoclast-specific marker expression (cellular proto-oncogene Fos, cathepsin K, nuclear factor of activated T-cells cytoplasmic 1, and TRAP) was analyzed via quantitative real-time polymerase chain reaction and Western blot. Bone resorption activity was evaluated using a calcium phosphate-coated pit assay. AMPK phosphorylation was examined to assess the associated signaling pathway. Results ADP-5 showed no effect on cell viability. It reduced TRAP-positive multinucleated cell formation and the mRNA and protein expression of key osteoclastogenic markers. Resorptive activity was significantly reduced in ADP-5-treated cells. In parallel, ADP-5 enhanced AMPK phosphorylation during differentiation. Conclusions ADP-5 suppressed RANKL-induced osteoclastogenesis and resorptive activity and was associated with enhanced AMPK phosphorylation. These findings identify ADP-5 as a potential modulator of RANKL-induced osteoclastogenesis and provide a rationale for further investigation of its effects and underlying mechanisms in inflammatory root resorption models.
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