经典Wnt信号通路对hDPSCs增殖、迁移和成牙本质向分化的影响

doi:10.13701/j.cnki.kqyxyj.2020.09.014

口腔医学研究 ›› 2020, Vol. 36 ›› Issue (9): 861-865.DOI: 10.13701/j.cnki.kqyxyj.2020.09.014

经典Wnt信号通路对hDPSCs增殖、迁移和成牙本质向分化的影响

关孟莹, 何丽娜, 潘爽, 李艳萍, 刘会梅, 牛玉梅^*

哈尔滨医科大学附属口腔医院牙体牙髓病科黑龙江哈尔滨 150001

收稿日期:2020-01-15 出版日期:2020-09-28 发布日期:2020-09-15
通讯作者: *牛玉梅,E-mail: niuym＠hrbmu.edu.cn
作者简介:关孟莹(1993~),女,黑龙江人,硕士,住院医师,主要从事牙髓干细胞再生的相关研究。
基金资助:
黑龙江省博士后基金(编号:LBH-Z17177);中国博士后基金(编号:2018M641871)

Effects of Canonical Wnt Signaling Pathway on Proliferation, Migration, and Odontogenic Differentiation of hDPSCs

GUAN Mengying, HE Lina, PAN Shuang, LI Yanping, LIU Huimei, NIU Yumei^*

Department of Endodontics, The First Affiliated Hospital of Harbin Medical University, Harbin 150001, China

Received:2020-01-15 Online:2020-09-28 Published:2020-09-15

摘要/Abstract

摘要： 目的: 探究经典Wnt/β-catenin信号通路对人牙髓干细胞(hDPSCs)增殖、迁移、成牙本质向分化及矿化的影响。方法: 筛选DKK1抑制Wnt信号通路的最佳浓度,并用该浓度处理hDPSCs。通过MTT和划痕实验观察细胞增殖和迁移能力的变化。碱性磷酸酶染色实验及Western Blot分析DKK1对hDPSCs成牙本质向分化能力的影响。茜素红染色探究DKK1对hDPSCs矿化前期阶段矿化能力的影响。结果: (1)MTT实验结果显示,DKK1组在第5天时,hDPSCs的A值高于对照组(P<0.05)。(2)划痕实验结果显示,细胞从划痕边界不断向中间迁移,在24 h时DKK1组细胞迁移距离明显大于对照组。(3)茜素红染色结果观察到,DKK1组可见多个大小不等的矿化结节。(4)Western blot结果显示,DKK1上调DMP-1的蛋白表达。结论: 抑制经典Wnt信号通路能够促进hDPSCs的增殖、迁移、成牙本质向分化及矿化。

关键词: Dikkopf-1, 人牙髓干细胞, 增殖, 迁移, 成牙本质向分化

Abstract: Objective:To investigate the effects of canonical Wnt/β-catenin signaling pathway on proliferation, migration, odontogenic differentiation, and mineralization of human dental pulp stem cells (hDPSCs). Methods: The optimal concentration of DKK1 was chosen to inhibit Wnt signaling pathway and treated hDPSCs. The proliferation and migration of hDPSCs were determined by MTT and scratching assay. The effects of DKK1 on odontogenic differentiation of hDPSCs were detected by Western Blot and ALP staining. Alizarin red was carried out to explore the effects of DKK1 on the mineralization ability of hDPSCs at the early stage of mineralization. Results: MTT test showed that the A of hDPSCs in the DKK1 group was higher than that in the control group on the 5th day(P<0.05). Scratching assay showed that the cells migrated from the scratch edge to the middle, and after 24h the cell migration distance of the DKK1 group was significantly larger than that of control group. Alizarin red staining showed that multiple mineralized nodules at various sizes were observed in the DKK1 group. Western blot results showed that DKK1 up-regulated the protein expression of DMP-1. Conclusion: Inhibiting canonical Wnt signaling pathway promoted the proliferation, migration, odontogenic differentiation, and mineralization of hDPSCs.

Key words: Dikkopf-1, human dental pulp stem cells, proliferation, migration, odontogenic differentiation

关孟莹, 何丽娜, 潘爽, 李艳萍, 刘会梅, 牛玉梅. 经典Wnt信号通路对hDPSCs增殖、迁移和成牙本质向分化的影响[J]. 口腔医学研究, 2020, 36(9): 861-865.

GUAN Mengying, HE Lina, PAN Shuang, LI Yanping, LIU Huimei, NIU Yumei. Effects of Canonical Wnt Signaling Pathway on Proliferation, Migration, and Odontogenic Differentiation of hDPSCs[J]. Journal of Oral Science Research, 2020, 36(9): 861-865.

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经典Wnt信号通路对hDPSCs增殖、迁移和成牙本质向分化的影响

Effects of Canonical Wnt Signaling Pathway on Proliferation, Migration, and Odontogenic Differentiation of hDPSCs

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