Vitamin C-linker-conjugated tripeptide AHK stimulates BMP-2-induced osteogenic differentiation of mouse myoblast C2C12 cells

Differentiation. May-Jun 2018;101:1-7. doi: 10.1016/j.diff.2018.03.001. Epub 2018 Mar 15.


Vitamin C-linker-conjugated Ala-His-Lys tripeptide (Vit C-AHK) is a derivative of Vitamin C-conjugated tripeptides, which were originally developed as a component of a product for collagen synthesis enhancement or human dermal fibroblast growth. Here, we investigated the effect of Vit C-AHK on bone morphogenetic protein (BMP)-2-induced osteoblast differentiation in a cell culture model. Vit C-AHK enhanced proliferation of C2C12 cells and induction of BMP-2-induced alkaline phosphatase, a typical marker of osteoblast differentiation. Vit C-AHK also stimulated the phosphorylation and translocation of Smad1/5/8 to the nucleus and phosphorylation of mitogen-activated protein kinases (MAPKs) including ERK1/2 and p38. In addition, Vit C-AHK enhanced the BMP-2-induced mRNA expression of osteoblast differentiation-related genes such as ALP, BMP-2, Osteocalcin, and Runx2. Our results suggest that Vit C-AHK exerts an enhancing effect on osteoblast proliferation and differentiation through activation of Smad1/5/8 and MAPK ERK1/2 and p38 signaling and without significant cytotoxicity. These results provide important data for the development of peptide-based bone-regenerative agents and treatment of bone-related disorders.

Keywords: ALP; BMP-2; MAPK; Osteoblast differentiation; Smad1/5/8; Vit C-AHK.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Ascorbic Acid / chemistry
  • Ascorbic Acid / pharmacology*
  • Bone Morphogenetic Protein 2 / metabolism*
  • Cell Differentiation / drug effects*
  • Cell Line
  • Mice
  • Myoblasts / cytology
  • Myoblasts / drug effects*
  • Osteoblasts / cytology
  • Osteocalcin / metabolism
  • Osteogenesis
  • Transforming Growth Factor beta / metabolism


  • Bmp2 protein, mouse
  • Bone Morphogenetic Protein 2
  • Transforming Growth Factor beta
  • Osteocalcin
  • Ascorbic Acid