Ark Shell Protein‐derived Bioactive Peptides Promote Osteoblastic Differentiation Through Upregulation of the Canonical Wnt/β‐catenin Signaling in Human Bone Marrow‐derived Mesenchymal Stem Cells

In this study, the stimulating effect of ark shell protein-derived peptides AWLNH and PHDL on osteoblast differentiation in human bone marrow-derived mesenchymal stem cells (hBMMSCs) and its molecular mechanism was investigated. The hBMMSCs were cultured with two peptides and osteogenic markers were analyzed. Results showed that enhanced ALP activity and calcification were detected in the presence of AWLNH and PHDL. Based on western blotting, RT-qPCR, and immunostaining analysis, AWLNH and PHDL are specific for osteoblast differentiation of hBMMSCs through activating the canonical Wnt/β-catenin signaling pathway followed by activating Runx2, osterix, and type I collagen. Loss-of-function assay with DKK-1, a Wnt antagonist, showed that the canonical Wnt/β-catenin signaling was essential for AWLNH and PHDL-induced osteogenesis in hBMMSCs. These findings suggested that AWLNH and PHDL can stimulate osteoblast differentiation of hBMMSCs via upregulating the canonical Wnt/β-catenin signaling and may be useful for a potential nutraceuticals or pharmaceuticals to treat osteoporosis. PRACTICAL APPLICATIONS: Ark shell is a popular foodstuff in Korea. However, biological effects of its protein and peptide have not been explored in many ways. This study demonstrated that ark shell protein-derived peptides promoted osteoblast differentiation in hBMMSCs through upregulating the canonical Wnt/β-catenin signaling. The results of this study could be a basis to promote its application as functional foods and/or nutraceuticals.