The effectiveness of asiaticoside on osteogenic differentiation ability in aging human periodontal ligament cells in vitro

Abstract

Retaining teeth is essential for the quality of life of the aging population. However, aging has biological effects in periodontal ligament cells with significant changes in their proliferative rate and regenerative function. Asiaticoside, extracted from Centella asiatica, was found to have potential in osteogenic differentiation and mineralization in human periodontal ligament cells (HPDLCs). The aim of this study is to investigate the effect of asiaticoside on the aging HPDLCs osteogenic differentiation and mineralization. Aging HPDLCs, retrieved from patients age 60 and above, were confirmed aging status using Senescence-Associated ß -Galactosidase assay. The cells were incubated with various concentrations of asiaticoside to test cell viability by MTT assay for 72 hours. After treating cells with asiaticoside for 1 and 7 days, the mRNA expression of osteogenic genes was analyzed by real-time polymerase chain reaction (PCR). Alizarin red was performed to evaluate the osteogenic differentiation and matrix mineralization. The results showed asiaticoside at concentrations 12.5 and 25 µM has no effect on cell viability and morphology. On day 1, the expression of BMP9, RUNX2 and OSX in 2.5 µM asiaticoside was significantly upregulated (P<0.05). Interestingly, DMP1 and BMP2 mRNA expressions in 25 µM asiaticoside were significantly enhanced at day 7 (P<0.05), while COL1 expression was decreased markedly. On day 14, Matrix mineralization was also significantly promoted (P<0.05). The results suggest asiaticoside can promote osteogenic differentiation in aging HPDLCs.