CGRP-modulated M2 macrophages regulate osteogenesis of MC3T3-E1 via Yap1

Bone fractures are one of the most frequent injuries in the musculoskeletal system. Despite the best treatment efforts, a large proportion of bone fracture cases still display undesirable outcomes. Here, we verified that calcitonin gene-related peptide (CGRP), a 37-amino acid neuropeptides, might be a critical regulator that link the nervous, immune and skeletal systems during bone healing. We used a CGRP overexpression lentiviral system and stably transfected M2 macrophages. Then, we investigated the biological function and the intrinsic mechanisms of CGRP on M2 macrophages. We confirmed that CGRP downregulated osteogenic factors (BMP2, BMP6, WNT10b and OSM) secretion at first and promoted them late on (p < 0.05). In addition, we utilized an indirect coculture system and further ascertain the influences of CGRP-induced M2 macrophages on MC3T3 osteogenesis. The results implied that CGRP-modulated osteoimmune environment elicit multiple effects on osteogenesis of MC3T3 during the entire observation period. Notably, verteporfin, a yes-associated protein 1 (Yap1) inhibitor, impaired CGRP effects significantly in our experiments. Taken together, our findings illustrated that CGRP might regulate osteogenesis by modulating the osteoimmune response of M2 macrophages via Yap1.