Effects Of Naringin On The Proliferation And Osteogenic Differentiation Of Human Periodontal Ligament Stem Cell In Vitro And In Vivo

Naringin is traditional Chinese medicine effective extracts of Drynaria, with clear chemical structure, which the role of the clinical efficacy have been verified. Naringin is confirmed promoting the process of bone marrow mesenchymal stem cells transform into osteoblasts. In recent years, traditional Chinese medicine treatment of periodontal disease as a supplement to conventional treatment in the clinic is taking more and more attention. This study was designed to screen out the optimal concentration of naringin effect on human periodontal ligament stem cells (hPDLSCs) and to study their osteogenic capability in the role of the optimal concentration.Materials and methods:The primary hPDLSCs were identified using immunofluorescence; on this basis, using Alizarin Red staining and Oil Red O staining to confirmed that the hPDLSCs have the ability to differentiate multiple; Measurement of alkaline phosphatase(ALP) activity to study naringin on cell of proliferation and osteogenic differentiation, screening the best effect concentration; By using RT-PCR expression of osteocalcin(OCN), osteopontin(OPN) and other bone-related genes to analyze gene expression at the optimum concentration of naringin; Finally, the cell composited with the nano-hydroxyapatite(n-HA) was transplanted into immunodeficient mice to explore its osteogenic potential in vivo.Results:Calcium nodules and orange oil droplets were observed under an optical microscope with Alizarin Red and Oil Red O staining; MTT, ALP showed that concentrations have a positive correlation with cell proliferation and differentiation when at low density, but the high groups will produce inhibition on cells, 1μM naringin on the proliferation and differentiation of hPDLSCs play a best role in promoting; RT-PCR showed that a bone-related genes had significantly higher expression compared to control groups; in vivo, apparent trabecular structure was observed by Hematoxylin-eosin(H-E) staining two months later, surrounded with a large amount of osteoblasts.Conclusion:Compared to the control group, 1μM naringin can promote the proliferation and differentiation of hPDLSCs; in the role of naringin, RUNX2, COL1A2, OPN, OCN expressed at high levels; within a certain period of time, hPDLSCs can be formed more trabecular bone, the bone maturation is also higher compared to the control group in vivo.