Abstract
BackgroundPlatelet-rich plasma (PRP) has the potential to be used for bone regeneration. However, its effect on osteogenic differentiation of human periodontal ligament stem cells (hPDLSCs) and its effect on cell autophagy of hPDLSCs remain unknown. In this study, we investigated the effects of PRP on cell viability and osteogenic differentiation of hPDLSCs and the underlying molecular mechanisms.MethodshPDLSCs were isolated and identified by morphology and flow cytometry analysis. Next, thrombin-activated PRP was used to stimulate hPDLSCs. The MTT assay was used to analyze cell viability. Osteogenic differentiation was investigated using alkaline phosphatase (ALP) activity assay, alizarin red S (ARS) staining, and gene expression analysis of osteogenic markers. Expression of the autophagic proteins was determined using western blotting.ResultsThrombin-activated PRP significantly enhanced cell viability, ALP activity, osteogenic-related mRNA levels and alizarin red-mineralization activity in hPDLSCs in a dose-dependent manner. Furthermore, activated PRP dose-dependently increased LC3-II/I ratio and the expression of SIRT1 and Beclin-1. PRP treatment also enhanced the autophagic flux. It was also demonstrated that the inhibition of SIRT1 using sirtinol or suppression of autophagy by 3-methyladenine (3-MA) abrogated PRP-induced viability and osteogenic differentiation of hPDLSCs.ConclusionOur study suggested that thrombin-activated PRP accelerated the viability and osteogenic differentiation of hPDLSCs via SIRT1-mediated autophagy induction.
Highlights
Periodontal ligament stem cells (PDLSCs) are multipotent mesenchymal stem cells (MSCs) derived from periodontal tissues, with the potentials of high proliferation, Xu et al Eur J Med Res (2021) 26:105 self-renewal and multidirectional differentiation [1]
Platelet-rich plasma (PRP) releases a large number of growth factors, such as platelet-derived growth factor (PDGF), transforming growth factor beta (TGF-β), insulin-like growth factor (IGF), epidermal growth factor (EGF) and vascular endothelial growth factor (VEGF), which plays a crucial role in bone regeneration due to its potential to repair tendons, ligaments, skeletal muscles and cartilage [6, 7]
Activated PRP enhances the cell viability and osteogenic differentiation of human periodontal ligament stem cells (hPDLSCs) To determine the potential effects of PRP on cell viability and osteogenic differentiation, we treated hPDLSCs with 5%, 10% and 20% thrombin-activated PRP
Summary
Periodontal ligament stem cells (PDLSCs) are multipotent mesenchymal stem cells (MSCs) derived from periodontal tissues, with the potentials of high proliferation, Xu et al Eur J Med Res (2021) 26:105 self-renewal and multidirectional differentiation [1]. PDLSCs have been proposed as alternative stem cells for periodontal regenerative therapy due to the osteogenic differentiation potential. Several researches have proposed that activated PRP can induce proliferation and osteogenic ability of bone marrow- and adiposederived mesenchymal stem cells and skeletal muscle satellite cells [10, 11]. We investigated the effects of PRP on cell viability and osteogenic differentiation of hPDLSCs and the underlying molecular mechanisms. Thrombin-activated PRP was used to stimulate hPDLSCs. The MTT assay was used to analyze cell viability. Osteogenic differentiation was investigated using alkaline phosphatase (ALP) activity assay, alizarin red S (ARS) staining, and gene expression analysis of osteogenic markers. Results: Thrombin-activated PRP significantly enhanced cell viability, ALP activity, osteogenic-related mRNA levels and alizarin red-mineralization activity in hPDLSCs in a dose-dependent manner. Conclusion: Our study suggested that thrombin-activated PRP accelerated the viability and osteogenic differentiation of hPDLSCs via SIRT1-mediated autophagy induction
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