Abstract
BackgroundPrevious research revealed that luteolin could improve the activation of alkaline phosphatase (ALP) and osteocalcin in mouse osteoblasts. We aimed to determine the effect of luteolin on osteogenic differentiation of periodontal ligament cells (PDLCs).MethodsCultured human PDLCs (HPDLCs) were treated by luteolin at 0.01, 0.1, 1, 10, 100 μmol/L, Wnt/β-catenin pathway inhibitor (XAV939, 5 μmol/L) alone or in combination with 1 μmol/L luteolin. Immunohistochemical staining was performed to ensure cells source. Cell activity and the ability of osteogenic differentiation in HPDLCs were determined by MTT, ALP and Alizarin Red S staining. Real-time Quantitative PCR Detecting System (qPCR) and Western blot were performed to measure the expressions of osteogenic differentiation-related genes such as bone morphogenetic protein 2 (BMP2), osteocalcin (OCN), runt-related transcription factor 2 (RUNX2), Osterix (OSX) and Wnt/β-catenin pathway proteins members cyclin D1 and β-catenin.ResultsLuteolin at concentrations of 0.01, 0.1, 1, 10, 100 μmol/L promoted cell viability, ALP activity and increased calcified nodules content in HPDLCs. The expressions of BMP2, OCN, OSX, RUNX2, β-catenin and cyclin D1 were increased by luteolin at concentrations of 0.01, 0.1, 1 μmol/L, noticeably, 1 μmol/L luteolin produced the strongest effects. In addition, XAV939 inhibited the expressions of calcification and osteogenic differentiation-related genes in HPDLCs, and 1 μmol/L luteolin availably decreased the inhibitory effect.Conclusion1 μmol/L luteolin accelerated osteogenic differentiation of HPDLCs via activating the Wnt/β-catenin pathway, which could be clinically applied to treat periodontal disease.
Highlights
As a common oral disease, periodontal disease is a main cause to tooth loss and could lead to local or even systemic effects [1]
Characteristics of human periodontal ligament cells (PDLCs) (HPDLCs) and effect of luteolin on cell activity The results of immunofluorescence detection showed that vimentin staining was positive, while keratin staining was negative in HPDLCs (Fig. 1a)
Effect of luteolin on osteogenic differentiation of HPDLCs The data in our study showed that different concentrations (0.01, 0.1, 1, 10, 100 μmol/L) of luteolin significantly enhanced alkaline phosphatase (ALP) activity (P < 0.001) and increased the content of calcified nodules in HPDLCs (P < 0.001)
Summary
As a common oral disease, periodontal disease is a main cause to tooth loss and could lead to local or even systemic effects [1]. Periodontal disease mainly promotes the regeneration of periodontal tissue, producing a certain number of healthy periodontal ligament cells (PDLCs) functioning as the primary basis for the repair of periodontal disease [2]. It has been reported that luteolin, which often exists as glycosylation in nature, could affect osteogenic differentiation [8]. Luteolin is a multifunctional complex that has a positive medicinal effect, for example, anti-cancer, anti-inflammatory, regulating immunity function, resisting oxidation and reducing osteoclast activity [11]. Previous research revealed that luteolin could improve the activation of alkaline phosphatase (ALP) and osteocalcin in mouse osteoblasts. We aimed to determine the effect of luteolin on osteogenic differentiation of periodontal ligament cells (PDLCs)
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