Baicalein enhances the osteogenic differentiation of human periodontal ligament cells by activating the Wnt/β-catenin signaling pathway

Abstract

This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain.  The optimal dosage of baicalein was 10 μM. DKK-1 significantly reversed the effects of baicalein on hPDLCs. The effects are exerted by activating the Wnt/β-catenin signaling pathway. Page 3 of 29A c c e p t e d M a n u s c r i p t ABSTRACT Objective: Periodontium regeneration is one of the most important processes for periodontitis therapy. Human periodontal ligament cells (hPDLCs) play a vital role in the repair and regeneration of periodontal tissues. Our study aimed to investigated the mechanisms underlying the promotion of hPLDCsosteogenic differentiation by baicalein. Design: hPDLCs were obtained from periodontal ligament (PDL) tissues by primary culture. The MTT assay was used to determine the growth curves of hPDLCs treated with different concentrations of baicalein (1.25, 2.5, 5, or 10 μM). Alkaline phosphatase (ALP) staining and Alizarin red S staining were performed to assess osteogenic differentiation of hPDLCs administered baicalein. Osteogenic differentiation-related gene and protein expression levels and Wnt/β-catenin pathway signal changes were assessed by qRT-PCR and Western blotting analysis. Results:The results showed that baicalein decreased the growth of hPDLCs slightly and increased ALP activity and calcium deposition in a dose-dependent manner. The expression of runt-related transcription factor 2 (RUNX2), bone morphogenetic protein 2 (BMP2), Osterix (OSX) and osteocalcin (OCN) were elevated after baicalein administration. Moreover, baicalein strongly activated the Wnt/β-catenin pathway and up-regulated the expression of β-catenin, lymphoid enhancer factor 1 (LEF1) and Cyclin D1. Dickkopf-related protein 1 (DKK-1) significantly reversed the effects of baicalein on hPDLCs. Conclusions: Our findings indicated that baicalein enhanced the osteogenic differentiation of hPDLCs via the activation of the Wnt/β-catenin signaling pathway, which may represent a potential candidate for periodontitis therapy.

TAZ contributes to osteogenic differentiation of periodontal ligament cells under tensile stress

“…PDLCs were obtained and cultured as previously described . Briefly, cells were identified through immunohistochemical staining for vimentin and cytokeratin (1:200, BosterBio).…”

“…Proteins were isolated, electrophoretically separated, and immunoblotted as previously described . Briefly, the membranes were incubated with the primary antibodies (1:1000, TAZ, 1:10000, COL I, Santa Cruz; 1:1000, Cbfα1, active β‐catenin, β‐catenin, LEF1, cyclin D1, β‐actin, Cell Signaling, USA; 1:2000, OSX, Abcam, UK; 1:1000, OCN, Merck; 1:10000, GAPDH, KangCheng), followed by HRP‐conjugated secondary antibodies.…”

“…PDLCs were obtained and cultured as previously described. 20 Briefly, cells were identified through immunohistochemical staining for vimentin and cytokeratin (1:200,BosterBio). The present study was approved by the Medical Ethics Committee of Hospital of Stomatology, Wenzhou Medical University, and informed consents were obtained from all participants prior to enrollment.…”

“…Proteins were isolated, electrophoretically separated, and immunoblotted as previously described. 20 Briefly, the membranes were…”

Effects of Different Concentrations of Crithmum Maritimum L. on Periodontal Ligament Fibroblast Cells

“…yaptıkları çalışmada, antioksidan kullanımının insan PDL hücrelerinde osteojenik farklılaşmayı artırdığını vurgulamışlardır. 12 KAYNAKLAR tonik, karbonhidrat, idrar söktürücü olarak veya obezite tedavisinde kullanılmaktadır. 14,15 Bununla birlikte Mekinić ve ark.…”