Zanthoxylum schinifolium enhances the osteogenic potential of periodontal ligament stem cells.

The present study demonstrates the osteogenic effect of Zanthoxylum schinifolium on periodontal ligament stem cells (PDLSCs). The dried herb of Z. schinifolium was first extracted with 70% ethanol and subsequently fractionated into five parts: n-hexane, methylene chloride (MC), ethyl acetate (EA), n-butanol (BuOH), and water fractions. The proliferation of PDLSCs was first assessed and increased by hexane, EA, or BuOH fraction of Z. schinifolium. We evaluated the osteogenic differentiation of PDLSCs by alkaline phosphatase (ALP) activity, messenger RNA (mRNA) expression of runt-related transcription factor 2 (RUNX2), osterix (OSX), FOSB, and FRA-1 as osteogenic transcription factors, and protein levels of osteopontin (OPN) and RUNX2 in response to each hexane, MC, EA, BuOH, or water fraction of Z. schinifolium. The significant ALP activity appeared in PDLSCs treated with hexane, EA, or BuOH fraction. The mRNA expression of osteogenic transcription factors was also increased by hexane, EA, or BuOH fraction with doses of 5, 10, 25, and 50 μg/ml compared to control group. We further assessed immunofluorescence staining with OPN and RUNX2 confirmed that the treatment of hexane, EA, or BuOH fraction enhances PDLSC osteogenic differentiation. In conclusion, these data suggest that fractions from Z. schinifolium differentially regulate PDLSC function. Among them, proliferation and osteogenic potential of PDLSCs were enhanced by hexane, EA, or BuOH fraction.