Rutin promotes osteogenic differentiation of periodontal ligament stem cells under inflammatory microenvironment

Abstract

To explore the effect of rutin on osteogenic differentiation of periodontal ligament stem cells under inflammatory microenvironment. Periodontal ligament stem cells (PDLSCs) were obtained by limited dilution method in vitro. PDLSCs were identified by flow cytometery. Lipopolysaccharide(LPS) was used to stimulate human periodontal ligament stem cells to establish an inflammation model in vitro. The experiment was divided into 4 groups: in group 1, only α-MEM was used to culture PDLSCs; in group 2, α-MEM medium containing LPS was used to culture PDLSCs, in group 3, rutin was added to α-MEM medium containing LPS to PDLSCs; and in group 4, α-MEM medium containing rutin was used to culture PDLSCs. Cell counting kit-8 was used to detect cell proliferation activity. Alkaline phosphatase(ALP) staining, ALP activity test, alizarin red staining, RT-PCR, and Western blot were used to detect the changes of osteogenic differentiation ability. The data were analyzed by SPSS 17.0 software package. The results of CCK-8 and ALP activity analysis showed that rutin at 10 μmol/L could significantly promote the proliferation and differentiation of periodontal stem cells under inflammatory state. ALP staining and alizarin red staining proved that (10 μmol/L) rutin could improve osteogenic differentiation of periodontal ligament stem cells under inflammatory microenvironment. RT-PCR and Western blot results showed that rutin could enhance the expression of osteogenic genes and proteins such as COL1, ALP, and RUNX2 under inflammatory state. Rutin can promote osteogenic differentiation of periodontal ligament stem cells under inflammatory microenvironment.