Sustained calcium ion release from bioceramics promotes CaSR-mediated M2 macrophage polarization for osteoinduction

Innate immune cells, especially macrophages, play a dual role in tissue repair and the defense against foreign bodies. Although biphasic calcium phosphate (BCP) ceramics have been confirmed as an excellent osteoimmunoregulatory biomaterial, it is unclear whether the ions release of BCP directly affects macrophage polarization and the mechanism by which the ions release is involved in osteoimmunomodulation. Herein, we verified the superior osteoinductive capacity of BCP in wild-type mice and showed its inability to promote this process in macrophage-deficient (LysM^-/- ) mice. Moreover, scanning electron microscopy, ion release curve, and calcein AM-staining results confirmed that BCP-released Ca²⁺ in a sustained manner, thereby maintaining the long-term induction of M2 macrophage polarization and promoting the differentiation of mesenchymal stem cells into osteoblasts during osteogenesis. Furthermore, Ca²⁺ targeted the Wnt/β-catenin signaling pathway and activated Arg1 and IL-10 (M2 marker genes) transcription through the calcium-sensing receptor (CaSR) in macrophages. Under treatment with a CaSR antagonist, macrophages cultured with the BCP fluid extract exhibited lower Ca²⁺ intake and weaker M2 macrophage polarization. These findings underscore the critical role of macrophages in bone regeneration and clarify the molecular mechanisms of Ca²⁺ -mediated osteoinduction by biomaterials, which is of great significance for the future design of biomaterial-oriented tissue regeneration engineering.