Effects of icariin on osteogenic differentiation of bone marrow stromal cells in beagle canine

Icariin (ICA), as the main active component in perennial herb Epimedium, displays a wide range of pharmacological and biological effects, including enhanced sexual function, immune regulation, anti-inflammatory, and anti-osteoporosis. In this study, the effect of ICA on osteogenic differentiation of bone marrow derived stromal cells (BMSCs) from a Beagle dog was assessed. BMSCs were Isolated from the canine bone marrow, cultured and identified by applying morphology and multi-lineage differentiation assays. Cell counting kit-8 (CCK-8) assay showed that ICA significantly promoted BMSC proliferation in a dose-dependent manner. Alkaline phosphatase (ALP) activity assay showed that ICA can increase ALP activity in a dose-dependent manner. Furthermore, RT-PCR and Western-blot indicated that ICA upregulated osteogenic-related genes and proteins. Alizarin red staining demonstrated the induced calcium deposition in BMSCs treated with 10^-6 mol/L ICA for 21 days. Overall, these findings clearly indicate that ICA is able to promote the osteogenic differentiation of BMSCs in beagle dogs. The optimal ICA concentration of 10^-6 mol/L may be appliable in bone tissue engineering.