神经调控cathepsin K阳性细胞谱系分化影响小鼠牙槽骨发育和稳态的研究

doi:10.3969/j.issn.1006-7795.2025.06.004

首都医科大学学报

神经调控cathepsin K阳性细胞谱系分化影响小鼠牙槽骨发育和稳态的研究

徐若诗^1,2，彭郑英¹，王玉顺¹，李轩^1,3，皮怀瑾^1,3，袁泉^1,3*

1. 口腔疾病防治全国重点实验室国家口腔医学中心国家口腔疾病临床医学研究中心，成都 610041；2. 四川大学华西口腔医院牙体牙髓病科，成都 610041；3. 四川大学华西口腔医院口腔种植科，成都 610041

收稿日期:2025-07-08 修回日期:2025-08-11 出版日期:2025-12-19 发布日期:2025-12-19
通讯作者: 袁泉 E-mail:yuanquan@scu.edu.cn
基金资助:
国家自然科学基金项目(82125006, 82001001), 口腔健康北京实验室开放项目(202301)，四川省科技计划项目(2024YFFK0134)。

The mechanism of neural modulation on the lineage differentiation of cathepsin K-positive cells in mouse alveolar bone development and homeostasis

Xu Ruoshi ^1,2, Peng Zhengying¹, Wang Yushun¹, Li Xuan^1,3, Pi Huaijin^1,3, Yuan Quan ^1,3*

1.State Key Laboratory of Oral Diseases & National Center for Stomatology & National Clinical Research Center for Oral Diseases, Chengdu 610041, China; 2. Department of Cariology and Endodontics, West China Hospital of Stomatology, Sichuan University , Chengdu 610041, China; 3. Department of Oral Implantology, West China Hospital of Stomatology, Sichuan University, Chengdu 610041, China

Received:2025-07-08 Revised:2025-08-11 Online:2025-12-19 Published:2025-12-19
Supported by:
This study was supported by National Natural Science Foundation of China (82125006, 82001001), Open Project with Beijing Laboratory of Oral Health (202301), Sichuan Science and Technology Program (2024YFFK0134).

摘要/Abstract

摘要： 目的通过组织蛋白酶K (cathepsin K, Ctsk) 阳性骨骼干细胞谱系示踪，以明确牙槽骨的骨组织、牙周膜和牙本质发育的细胞分化和外周神经调控机制。方法首先构建Ctsk-Cre;tdTomato;Col2.3-GFP荧光报告小鼠模型，收取新生小鼠牙槽骨骨组织、牙周膜组织和牙髓组织，进行Ctsk阳性谱系示踪和免疫荧光染色。进一步构建小鼠下牙槽神经切断模型，分析tdTomato⁺Col2.3-GFP⁺双阳性细胞。结果本研究成功构建Ctsk-Cre;tdTomato;Col2.3-GFP荧光报告小鼠模型。新生小鼠谱系示踪和免疫荧光染色显示，牙槽骨tdTomato阳性细胞表达含锌指成骨细胞特异性转录因子 (osterix，Osx)，牙周膜tdTomato阳性细胞表达Periostin，牙本质tdTomato阳性细胞表达Col2.3-GFP。与神经完整组相比，失神经小鼠的tdTomato阳性谱系减少，成分化状态的tdTomato⁺Col2.3-GFP⁺双阳性细胞减少。结论新生小鼠牙槽骨发育阶段，Ctsk阳性细胞参与骨组织、牙周膜和牙本质的发育，并受下牙槽神经的调控，对成年损伤修复和稳态维持具有重要意义。

关键词: 谱系示踪, 基因小鼠模型, 荧光, 发育, 神经, 牙本质

Abstract: Objective To explorethe cellular differentiation mechanisms, with Cathepsin K (Ctsk)-positive skeletal stem cells (SSCs) lineage tracing, underlying alveolar bone tissue, periodontal ligament, and dentin development, as well as the regulatory role of peripheral nerves.Methods Ctsk-Cre;tdTomato;Col2.3-GFP dual-fluorescence mouse models were generated. Tissues including alveolar bone, periodontal ligament, and dental pulp were harvested from neonatal mice for Ctsk-positive lineage tracing and immunofluorescence staining. Futhermore, an inferior alveolar nerve transection model was established to analyze tdTomato⁺Col2.3-GFP⁺ double-positive cells.Results The Ctsk-Cre;tdTomato;Col2.3-GFP fluorescent reporter mouse model was successfully generated. Lineage tracing and immunofluorescence staining in neonatal mice revealed that tdTomato+ cells in the alveolar bone expressed osterix, tdTomato+ cells in the periodontal ligament expressed Periostin, and tdTomato+ cells in the dentin expressed Col2.3-GFP. Further analysis with the inferior alveolar nerve transection model showed reduced tdTomato+ lineage and decreased numbers of differentiating tdTomato⁺ Col2.3-GFP⁺ double-positive cells in denervated mice compared to the nerve-intact group.Conclusions During the developmental stage of alveolar bone in neonatal mice, Ctsk positive cells participated in the development of bone tissue, periodontal ligament, and dentin, which were regulated by the inferior alveolar nerve. It shed light on injury repair and homeostasis of adulthood skeleton.

Key words: lineage tracing, genetic mouse model, fluorescence, development, nerve, dentin

中图分类号:

Q38，Q344

徐若诗, 彭郑英, 王玉顺, 李轩, 皮怀瑾, 袁泉. 神经调控cathepsin K阳性细胞谱系分化影响小鼠牙槽骨发育和稳态的研究[J]. 首都医科大学学报, doi: 10.3969/j.issn.1006-7795.2025.06.004.

Xu Ruoshi , Peng Zhengying, Wang Yushun, Li Xuan, Pi Huaijin, Yuan Quan. The mechanism of neural modulation on the lineage differentiation of cathepsin K-positive cells in mouse alveolar bone development and homeostasis[J]. Journal of Capital Medical University, doi: 10.3969/j.issn.1006-7795.2025.06.004.

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神经调控cathepsin K阳性细胞谱系分化影响小鼠牙槽骨发育和稳态的研究

The mechanism of neural modulation on the lineage differentiation of cathepsin K-positive cells in mouse alveolar bone development and homeostasis

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