Abstract
Objective: This study aimed to investigate the roles of long noncoding RNA (lncRNA) maternally expressed gene 3 (MEG3) in osteogenic differentiation of periodontal ligament stem cells (PDLSCs) in periodontitis.Methods: Differentially expressed lncRNAs and mRNAs between periodontitis periodontal ligament tissues and healthy periodontal ligament tissues were selected out using R project. PDLSCs were identified using flow cytometry. Western blot was employed to detect pathway relative proteins. Besides, targeted relationships between lncRNA and miRNA, as well as miRNA and mRNA were verified by dual luciferase reporter gene assay. Osteogenic differentiation of PDLSCs was assessed by alkaline phosphatase (ALP) staining and Alizarin Red Staining (ARS). Markers for osteoblast (Runx2, Osterix, Osteocalcin, Colla1) were detected using western blot.Results: LncRNA MEG3 and IGF1 were both down-regulated, while miR-27a-3p was up-regulated in periodontitis samples compared with healthy samples. Overexpression of MEG3 promoted osteogenic differentiation by enhancing expression of IGF1 yet suppressing expression of miRNA-27a-3p. Meanwhile, the results of ALP and ARS staining indicated that up-regulation of lncRNA MEG3 or IGF1 promoted osteogenic differentiation in PDLSCs, which could be reversed with up-regulation of miRNA-27a-3p.Conclusion: Down-regulation of MEG3 suppressed osteogenic differentiation of PDLSCs through miR-27a-3p/IGF1 axis in periodontitis.
Highlights
Periodontitis is a widespread infectious human chronic inflammatory disease which is characterized by progressive disruption of the connective tissue surrounding the teeth and the inflammatory reaction mostly caused by an oral microbial biofilm [1]
IGF1 down-regulated in phosphatidylinositol 3kinase (PI3K)/Akt signaling pathway could be inactivated in periodontitis periodontal tissues compared with healthy periodontal tissues
LncRNA maternally expressed gene 3 (MEG3) was down-regulated in periodontitis periodontal tissues compared with healthy periodontal tissues and correlated with IGF1 via miR-27a-3p
Summary
Periodontitis is a widespread infectious human chronic inflammatory disease which is characterized by progressive disruption of the connective tissue surrounding the teeth and the inflammatory reaction mostly caused by an oral microbial biofilm [1]. Serious periodontitis infects 9% to 11% of adult population in the world, especially in adult population over 50 years of age [2,3,4]. Periodontal ligament stem cells (PDLSCs) are a group of mesenchymal stem cells (MSCs) derived from the periodontal ligament, which show multiple differentiation capability and a promising potential of being used to regenerate supporting tissues for the patients recently [6, 7]. Current researches have demonstrated that activating phosphatidylinositol 3kinase (PI3K) / Akt signaling pathway could regulate the osteogenic differentiation of PDLSCs [8]. Though most of the studies focus on cancers, there are still some of the researches for periodontitis [9,10,11]
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