MaoHua Meng1,2, Qian Xia3, Ying Li1,2, Xin Chen1,2, QinYing Wang1,2, JingQiao Chen1,2, XingXing Xu1,2, Huan Wang1,2, JiaYu Shu1,2, Jing Lu1,2, Lu Cheng4, ZhaoYang Ye5, Bin Song6, Qiang Dong7,8. 1. School of Stomatology, Guizhou Medical University, 9 Beijing Road, Yunyan District, Guiyang City, 550004, Guizhou Province, China. 2. Department of Prosthodontics, Stomatological Hospital of Guizhou Medical University, 9 Beijing Road, Yunyan District, Guiyang City, 550004, Guizhou Province, China. 3. Department of Preventive and Pediatric Dentistry, Stomatological Hospital of Guizhou Medical University, Guiyang City, 550004, Guizhou Province, China. 4. Department of Stomatology, Guiyang Hospital of Stomatology, Guiyang City, 550005, Guizhou Province, China. 5. Clinical Research Center, The Affiliated Hospital of Guizhou Medical University, 9 Beijing Road, Yunyan District, Guiyang City, 550004, Guizhou Province, China. yezy@gmc.edu.cn. 6. Department of Prosthodontics, Guizhou Provincial People's Hospital, 83 Zhongshan East Road, Guiyang, Guiyang City, 550003, Guizhou Province, China. songbin0407@163.com. 7. School of Stomatology, Guizhou Medical University, 9 Beijing Road, Yunyan District, Guiyang City, 550004, Guizhou Province, China. dongstrongvv@outlook.com. 8. Department of Prosthodontics, Stomatological Hospital of Guizhou Medical University, 9 Beijing Road, Yunyan District, Guiyang City, 550004, Guizhou Province, China. dongstrongvv@outlook.com.
Abstract
INTRODUCTION: The influence of enamel matrix derivative (EMD) on proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) was explored in high glucose (HG) microenvironment with interaction of Wnt/β-catenin pathway. MATERIALS AND METHODS: Extraction of BMSCs from Sprague-Dawley rats, culture, and identification were manifested. The cells were treated with different concentration of EMD in HG to figure out the most available concentration for proliferation and osteogenic differentiation. Then, observation of cell growth curve and cell cycle changes, and detection of Osterix, runt-related transcription factor 2 (Runx2), COL-I, early osteogenic indexes, Calcium salt deposition, and β-catenin protein in Wnt/β-catenin pathway were assured. After adding Wnt/β-catenin pathway inhibitor (XAV-939) in the cells with osteogenesis induction, detection of binding of β-catenin to Osterix was clarified. RESULTS: Via identification BMSCs cultured in vitro was qualified. Different concentrations of EMD could accelerate cell proliferation in HG and osteogenesis induction, and 75 μg/mL EMD had the best effect. The HG augmented BMSCs proliferation and the propidium iodide index of flow cytometry cycle was elevated in HG, which were strengthened via the EMD. After BMSCs' osteogenesis induction, Osterix, Runx2, CoL-1, early osteogenic indexes, and calcium salt deposition were reduced, but elevated via EMD. β-Catenin was the lowest in the HG, but elevated after EMD. After addition of XAV-939, reduction of β-catenin and the downstream (Osterix and Runx2) were manifested. Detection of binding protein bands was in β-catenin and Osterix of the HG after EMD treatment. CONCLUSION: EMD may facilitate the osteogenic differentiation of BMSCs via activating the Wnt/β-catenin pathway in HG.
INTRODUCTION: The influence of enamel matrix derivative (EMD) on proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) was explored in high glucose (HG) microenvironment with interaction of Wnt/β-catenin pathway. MATERIALS AND METHODS: Extraction of BMSCs from Sprague-Dawley rats, culture, and identification were manifested. The cells were treated with different concentration of EMD in HG to figure out the most available concentration for proliferation and osteogenic differentiation. Then, observation of cell growth curve and cell cycle changes, and detection of Osterix, runt-related transcription factor 2 (Runx2), COL-I, early osteogenic indexes, Calcium salt deposition, and β-catenin protein in Wnt/β-catenin pathway were assured. After adding Wnt/β-catenin pathway inhibitor (XAV-939) in the cells with osteogenesis induction, detection of binding of β-catenin to Osterix was clarified. RESULTS: Via identification BMSCs cultured in vitro was qualified. Different concentrations of EMD could accelerate cell proliferation in HG and osteogenesis induction, and 75 μg/mL EMD had the best effect. The HG augmented BMSCs proliferation and the propidium iodide index of flow cytometry cycle was elevated in HG, which were strengthened via the EMD. After BMSCs' osteogenesis induction, Osterix, Runx2, CoL-1, early osteogenic indexes, and calcium salt deposition were reduced, but elevated via EMD. β-Catenin was the lowest in the HG, but elevated after EMD. After addition of XAV-939, reduction of β-catenin and the downstream (Osterix and Runx2) were manifested. Detection of binding protein bands was in β-catenin and Osterix of the HG after EMD treatment. CONCLUSION: EMD may facilitate the osteogenic differentiation of BMSCs via activating the Wnt/β-catenin pathway in HG.
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