Nell-1蛋白通过Wnt/β-catenin通路调控人上颌窦黏膜来源间充质干细胞成骨分化

doi:10.13701/j.cnki.kqyxyj.2023.03.009

口腔医学研究 ›› 2023, Vol. 39 ›› Issue (3): 227-235.DOI: 10.13701/j.cnki.kqyxyj.2023.03.009

Nell-1蛋白通过Wnt/β-catenin通路调控人上颌窦黏膜来源间充质干细胞成骨分化

徐静¹, 吕慧欣¹, 王一涵¹, 鲍鑫², 张逸¹, 周延民^1*

1.吉林大学口腔医院种植科,吉林省牙发育及颌骨重塑与再生重点实验室吉林长春 130012;
2.吉林大学口腔医院修复科吉林长春 130012

收稿日期:2022-10-27 出版日期:2023-03-28 发布日期:2023-03-21
通讯作者: * 周延民,E-mail:zhouym@jlu.edu.cn
作者简介:徐静(1995~ ),女,山东临沂人,硕士在读,主要从事口腔种植临床与基础研究。
基金资助:
国家自然科学基金面上项目(编号:82071152)

Nell-1 Regulates Osteogenic Differentiation of Human Maxillary Sinus Membrane-derived Mesenchymal Stem Cells through Wnt/β-catenin Pathway

XU Jing¹, LV Huixin¹, WANG Yihan¹, BAO Xin², ZHANG Yi¹, ZHOU Yanmin^1*

1. Department of Implantology, Hospital of Stomatology, Jilin University, Jilin Provincial Key Laboratory of Tooth Development and Bone Remodeling, Changchun 130012, China;
2. Department of Prosthodontics, Hospital of Stomatology, Jilin University, Changchun 130012, China

Received:2022-10-27 Online:2023-03-28 Published:2023-03-21

摘要/Abstract

摘要： 目的: 研究不同浓度Nell-1(Nel-like type 1 molecule)蛋白对人上颌窦黏膜来源间充质干细胞(maxillary sinus membrane-derived mesenchymal stem cells, MSMSCs)成骨分化的影响。方法: 体外分离培养并鉴定人MSMSCs。使用Nell-1蛋白(10、100、1000 ng/mL)诱导MSMSCs,利用CCK-8法和细胞活死染色法检测细胞活性变化;碱性磷酸酶活性检测细胞成骨能力改变;实时定量荧光聚合酶链反应(qRT-PCR)检测成骨相关因子mRNA表达情况;免疫荧光染色和Western blot法检测成骨蛋白表达情况。结果: Nell-1蛋白对MSMSCs有一定的促增殖作用;碱性磷酸酶活性检测显示各实验组成骨效果优于对照组;qRT-PCR、免疫荧光染色和Western blot结果显示,100 ng/mL的Nell-1显著促进β-catenin、Runx-2、OCN等mRNA及蛋白的表达(P＜0.01)。结论: Nell-1蛋白可通过上调Wnt/β-catenin信号通路的表达提高MSMSCs体外成骨分化能力。

关键词: 间充质干细胞, 骨再生, 上颌窦, Nell-1

Abstract: Objective: To investigate the effect of Nell-1 protein on osteogenic differentiation of human maxillary sinus membrane-derived mesenchymal stem cells (MSMSCs). Methods: The MSMSCs were induced by Nell-1 protein (10,100,1000 ng/mL), and cell activity changes were detected using CCK-8 assay and cell live-dead staining. The changes in osteogenic ability were detected by alkaline phosphatase activity. mRNA expression of osteogenic-related factors was detected by quantitative real-time fluorescence PCR (qRT-PCR). Immunofluorescence staining and western blot were used to detect osteogenic protein expression. Results: Nell-1 protein had a promote effect on proliferative of MSMSCs. Alkaline phosphatase activity assay showed that the osteogenic effect of each experimental group was better than that of the control group. qRT-PCR, immunofluorescence staining, and western blot showed that 100 ng/mL of Nell-1 significantly promoted the mRNAs and protein expression of β-catenin, Runx-2, OCN, and others (P<0.01). Conclusion: Nell-1 protein can improve the osteogenic differentiation ability of MSMSCs in vitro by upregulating the expression of Wnt/β-catenin signaling pathway.

Key words: mesenchymal stem cells, bone regeneration, maxillary sinus, Nell-1

徐静, 吕慧欣, 王一涵, 鲍鑫, 张逸, 周延民. Nell-1蛋白通过Wnt/β-catenin通路调控人上颌窦黏膜来源间充质干细胞成骨分化[J]. 口腔医学研究, 2023, 39(3): 227-235.

XU Jing, LV Huixin, WANG Yihan, BAO Xin, ZHANG Yi, ZHOU Yanmin. Nell-1 Regulates Osteogenic Differentiation of Human Maxillary Sinus Membrane-derived Mesenchymal Stem Cells through Wnt/β-catenin Pathway[J]. Journal of Oral Science Research, 2023, 39(3): 227-235.

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Nell-1蛋白通过Wnt/β-catenin通路调控人上颌窦黏膜来源间充质干细胞成骨分化

Nell-1 Regulates Osteogenic Differentiation of Human Maxillary Sinus Membrane-derived Mesenchymal Stem Cells through Wnt/β-catenin Pathway

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