Synergistic effect of Wnt modulatory small molecules and an osteoinductive ceramic on C2C12 cell osteogenic differentiation

- THQ-1a and PP-9 were identified as novel osteogenic compounds that increased Runx2 and BMP2 expression in C2C12 cells.
- Osteogenic differentiation by THQ-1a and PP-9 was shown by enhanced ALP functional activity, Osteocalcin, Collagen I expression, and calcification.
- THQ-1a or PP-9 in the presence of an osteoinductive TCP synergizes osteogenesis as observed for ALP functional activity.
- Mechanistic studies showed that THQ-1a and PP-9 act by modulating Wnt signaling in C2C12 cells.

Although osteoinductive ceramics can induce osteoblast differentiation in vitro and bone regeneration in vivo, their effects rely solely on the limited number of endogenous stem cells. More recently, ceramic carriers seeded with culture-expanded stem cells have been reported as implants capable of in vivo bone formation. However, effective and safe signaling agents that promote cell differentiation to the osteogenic lineage are still needed. In the present report, two osteogenic small-molecules THQ-1a and PP-9 were identified by testing a series of compounds for Runx2 and BMP2 expression in C2C12 cells. Compounds THQ-1a and PP-9 modulated Wnt signaling and enhanced the expression of molecular markers of osteoblast differentiation. To probe the utility of these compounds for use with ceramic cell implants, the effect of THQ-1a and PP-9 on C2C12 cell osteogenic differentiation was investigated in the presence of a tricalcium phosphate (TCP) ceramic. The effect of THQ-1a and PP-9 on markers such as Osteocalcin and Collagen I was significantly increased in the presence of TCP ceramic. Additionally, THQ-1a or PP-9 in the presence of TCP ceramic gave a synergistic increase in alkaline phosphatase activity in the differentiation of C2C12 cells. Taken together, the results suggest an approach to directing cell lineage commitment for bone regeneration by the application of small-molecule osteogenic agents to cells in the presence of osteoinductive ceramics.

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