Silibinin is a secondary metabolite derived from Silybum marianum. Silibinin had been used as a hepatoprotective substance for more than 2,000 years in Europe. A number of studies have revealed the efficacy of silibinin as an antioxidant preventing skin damages, but there is a lack of study on the hair inductive property of silibinin. Thus, we assess the effectiveness of silibinin as a therapeutic candidate for alopecia using three-dimensional cultured human dermal papilla cells.
 In this study, cultured human dermal papilla cells were seeded into 96-well round-bottom microplate to make the unified spheres. After the spheres were treated with silibinin for 48 hours, the size of cultured human dermal papilla cells was calculated using phase-contrast microscopy. To assess the cell growth effect, AKT protein levels were evaluated using western blotting. Using luciferase reporter assay, the activity of Wnt/β-catenin signaling pathways associated with hair proliferation was analyzed. Quantative real time polimerase chain reaction assay was performed to evaluate upregulation of the signiture genes such as VCAN, FGF7, BMP2 and ALPL.
The results of our study revealed silibinin increases the diameter of three-dimensional cultured human dermal papilla cells and elevates AKT protein levels. And Wnt/β-catenin signaling pathway was activated remarkably in silibinin treated spheres. In addition, signature genes of human dermal papill cell such as VCAN, FGF7, BMP2, and ALPL were upregulated by silibinin treatment.
 In conclusion, silibinin enhances the signaling pathway related to hair proliferation. Our study suggests silibinin could be a successful therapeutic agent for alopecia.

Association between Hair Proliferation and Flavonoid Silibinin in Human Dermal Papilla Cells