Hedgehog Signaling Is Involved In Naringin Promoting Osteogenic Differentiation Of Human Periodontal Ligament Cells

Periodontal tissue includes gingiva,periodontal ligament,alveolar bone and cementum.The remodeling of it is the biological basis of orthodontic tooth movement(OTM).Many studies have confirmed that traditional Chinese medicine can affect the process of orthodontic tooth movement.Naringin,a traditional Chinese medicine,mainly exists in the pericarp and pulp of Rutaceae plants.It is one of the main effective components of Rhizoma Drynariae and has the ability to promote the proliferation and osteogenic differentiation of periodontal ligament cells.Hedgehog signaling pathway is involved in regulating bone growth and development.Runx2 plays an important role in Hedgehog promoting late stage of differentiation of osteoblasts.Naringin can promote the expression of Runx2 in BMSCs cultured in vitro.Therefore,we infer that naringin can promote orthodontic tooth movement by activating Hedgehog signaling pathway.Purpose:To find out the optimal concentration and action time of naringin in promoting osteogenic differentiation of hPDLSCs,and to explore the role of Hedgehog signaling in this promotion.Methods:Identify hPDLSCs by flow cytometry.Confirm that hPDLSCs have differentiation potential by using Alizarin Red staining and Oil Red O staining.Using quantitative test of alkaline phosphatase(ALP)to figure out the optimal concentration and action time of naringin.Determine the expression level of ALP?OCN?Runx2?SMO?Glil?Ptch by RT-PCR and Western Blotting.Results:1.Human periodontal ligament cells were successfully cultured by the method of tissue block,with normal shape and good conditions.Flow cytometry showed positive expression of CD44,CD90 and CD105,but negative expression of CD34 and CD45.Calcium nodules and orange oil droplets were observed in osteogenic and adipogenic induction.2.Measurement of alkaline phosphatase showed that naringin had the best osteogenic effect when 7 days with its concentration of lmg/L.3.Naringin can promote osteogenic differentiation of hPDLSCs through Hedgehog signaling pathway,after using cyclopamine,the expressions of ALP,Runx2,SMO,GLI1 and PTCH 1 were decreased.Conclusions:1.Naringin can promote the osteogenic differentiation of hPDLSCs.The relationship between the optimal concentration and the time of action was explored by ALP test.2.Hedgehog signaling pathway plays a regulatory role in osteogenic differentiation of hPDLSCs.3.Naringin can promote osteogenic differentiation of hPDLSCs through Hedgehog signaling pathway.