Bin Jiang1, Jie Xu1, Yifei Zhou2, Jie Mao3, Guangzhao Guan4, Xiaomei Xu1, Li Mei5. 1. Department of Orthodontics, Hospital of Stomatology, Orofacial Reconstruction and Regeneration Laboratory, Southwest Medical University, Luzhou. 2. Department of Stomatology, People's Hospital of Langzhong City, Nanchong. 3. Department of Stomatology, People's Hospital of Rongchang City, Chongqing, China. 4. Department of Oral Diagnostic and Surgical Sciences. 5. Department of Oral Science, Sir John Walsh Research Institute, Faculty of Dentistry, University of Otago, Dunedin, New Zealand.
Abstract
OBJECTIVES: 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). METHODS: 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. RESULTS: 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. CONCLUSIONS: E2 enhanced osteogenic differentiation of hPDLSCs through the activation of the Wnt/β-catenin signaling pathway.
OBJECTIVES: 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). METHODS: 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. RESULTS: 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. CONCLUSIONS:E2 enhanced osteogenic differentiation of hPDLSCs through the activation of the Wnt/β-catenin signaling pathway.