Asiaticoside Induces Matrix Synthesis and Osteogenic/Cementogenic Differentiation in HPDL Cells

Objective: Asiaticoside, an extract from Centrella asiatica, has been widely used to promote wound healing. In this study, the effects of asiaticoside on proliferation, protein synthesis and osteogenic/cementogenic differentiation in human periodontal ligament cells (HPDLs) were investigated. Methods: Human periodontal ligament (HPDL) cells were treated with different concentrations of asiaticoside (25, 50 and 100 μg/ml). Cell number was determined by MTT assay. The mRNA expression was analyzed by reverse transcription-polymerase chain reaction (RT-PCR). Western blot analysis and immunocytochemistry were used to confirm protein synthesis. Osteogenic/cementogenic differentiation was determined by alkaline phosphatase (ALP) activity, osteogenic genes expression and in vitro mineralization. Results: Asiaticoside (25 to 100 mg/ml) had neither effect on cytotoxicity nor cell proliferation. Increased expression of fibronectin (FN) and type I collagen (COL I) was observed at both mRNA and protein levels at 72 hours. Moreover, asiaticoside attenuated MMP-1 but enhanced TIMP-1 mRNA expression. The addition of asiaticoside in osteogenic medium resulted in the increase of ALP activity, the up-regulation of osteoblast/cementoblast marker gene mRNA expression and the enhancement of mineralization in HPDL cells. These results suggest the potential application of asiaticoside for enhancing the periodontal tissue healing. Conclusion: Asiaticoside could increase the fibronectin and type I collagen synthesis as well as promote the osteogenic/cementogenic differentiation of HPDL cells. The results indicate the feasible application of asiaticoside for periodontal tissue healing and regeneration. Grants: Higher Education Research Promotion and National Research University Project of Thailand, Office of the Higher Education Commission and Chulalongkorn University Centenary Academic Development Project.