Jiang Bin, Xu Jie, Zhou Yifei, Mao Jie, Guan Guangzhao, Xu Xiaomei, Mei Li
Department of Orthodontics, Hospital of Stomatology, Orofacial Reconstruction and Regeneration Laboratory, Southwest Medical University, Luzhou.
Department of Stomatology, People's Hospital of Langzhong City, Nanchong.
J Craniofac Surg. 2020 Mar/Apr;31(2):583-587. doi: 10.1097/SCS.0000000000006226.
This study was designed to investigate the role of the Wnt/β-catenin signaling pathway in estrogen-enhanced osteogenic differentiation of human peridontal ligament stem cells (hPLSCs).
The limiting dilution technique was used for cloning and purification of hPLSCs. Flow cytometric analysis of STRO-1, CD146, and CD45 was conducted to identify hPLSCs. The P3 hPDLSCs were divided into 4 groups: Control, 10M E2, 10M E2+100 ng/mL Wnt3a, 10M E2+5 × 10M Xav939. After 7 days of osteogenic induction, qRT-PCR was used to detect the mRNA expression of β-catenin, CyclinD1, alkaline phosphatase, Runx2, and OCN; Western blot was used to detect the protein expression of β-catenin, GSK3β, P-GSK3β, CyclinD1, Runx2, and OCN; After 1, 3, 5, 7 days of osteogenic induction, the activity of alkaline phosphatase was detected.
The authors' results showed that E2 was able to enhance the osteogenic differentiation of hPDLSCs and Wnt/β-catenin signaling pathway was involved. Wnt3a activated the signaling pathway of Wnt/β-catenin and further enhanced the osteogenesis of hPDLSCs. Xav939 inhibited the Wnt/β-catenin signaling pathway in estrogen-mediated environment, but did not obviously inhibit the osteogenic differentiation of hPDLSCs.
E2 enhanced osteogenic differentiation of hPDLSCs through the activation of the Wnt/β-catenin signaling pathway.
本研究旨在探讨Wnt/β-连环蛋白信号通路在雌激素增强人牙周膜干细胞（hPLSCs）成骨分化中的作用。
采用有限稀释技术对hPLSCs进行克隆和纯化。通过对STRO-1、CD146和CD45进行流式细胞术分析来鉴定hPLSCs。将第3代hPDLSCs分为4组：对照组、10 nM E2组、10 nM E2 + 100 ng/mL Wnt3a组、10 nM E2 + 5×10 nM Xav939组。成骨诱导7天后，采用qRT-PCR检测β-连环蛋白、细胞周期蛋白D1、碱性磷酸酶、Runx2和骨钙素的mRNA表达；采用蛋白质免疫印迹法检测β-连环蛋白、糖原合成酶激酶3β（GSK3β）、磷酸化糖原合成酶激酶3β（P-GSK3β）、细胞周期蛋白D1、Runx2和骨钙素的蛋白表达；成骨诱导1、3、5、7天后，检测碱性磷酸酶活性。
作者的结果表明，E2能够增强hPDLSCs的成骨分化，且Wnt/β-连环蛋白信号通路参与其中。Wnt3a激活Wnt/β-连环蛋白信号通路，进一步增强hPDLSCs的成骨作用。Xav939在雌激素介导的环境中抑制Wnt/β-连环蛋白信号通路，但未明显抑制hPDLSCs的成骨分化。
E2通过激活Wnt/β-连环蛋白信号通路增强hPDLSCs的成骨分化。
