上海交通大学学报（医学版）

上海交通大学学报（医学版） >

2025 , Vol. 45 >Issue 9: 1106 - 1115

DOI: https://doi.org/10.3969/j.issn.1674-8115.2025.09.003

论著 · 基础研究

颌骨成骨细胞调控B细胞分化的效应研究

  • 王歆雨 ,
  • 陈芊烨 ,
  • 孙计萍 ,
  • 鲁婷玮 ,
  • 黄湘如 ,
  • 孙思远 ,
  • 刘媛琪 ,
  • 潘厚文 ,
  • 代庆刚 ,
  • 沈蕾 ,
  • 江凌勇
展开
  • 1.上海交通大学医学院附属第九人民医院口腔颅颌面科，上海交通大学口腔医学院，国家口腔医学中心，国家口腔疾病临床医学研究中心，上海市口腔医学重点实验室，上海市口腔医学研究所，上海 200011
    2.上海交通大学医学院附属第九人民医院口腔颌面-头颈肿瘤科，上海交通大学口腔医学院，国家口腔医学中心，国家口腔疾病临床医学研究中心，上海市口腔医学重点实验室，上海市口腔医学研究所，上海 200011
    3.上海交通大学基础医学院免疫学与微生物学系，上海市免疫学研究所，上海 200025
    4.上海交通大学医学院附属第九人民医院口腔第二门诊部，上海交通大学口腔医学院，国家口腔医学中心，国家口腔疾病临床医学研究中心，上海市口腔医学重点实验室，上海市口腔医学研究所，上海 200011
江凌勇，主任医师，博士；电子信箱：jianglingyong@sjtu.edu.cn。

收稿日期: 2025-02-19

  录用日期: 2025-04-29

  网络出版日期: 2025-09-30

基金资助

上海交通大学医学院“双百人”项目(20221809);国家自然科学基金(82430032);国家自然科学基金(81870740);国家自然科学基金(82071083);国家自然科学基金(82271006);国家重点研发计划(2024YFC2510700);上海市科技创新行动计划国际科技合作项目/政府间国际科技合作项目(23410713600);上海市自然科学基金(21ZR1436900);上海市自然科学基金(22ZR1436700);上海交通大学医学院附属第九人民医院交叉研究基金(JYJC202116);上海交通大学医学院生物材料与再生医学交叉研究项目(2022LHB02)

收起

Effect of jaw osteoblasts on B cell development via cytokine secretion

  • WANG Xinyu ,
  • CHEN Qianye ,
  • SUN Jiping ,
  • LU Tingwei ,
  • HUANG Xiangru ,
  • SUN Siyuan ,
  • LIU Yuanqi ,
  • PAN Houwen ,
  • DAI Qinggang ,
  • SHEN Lei ,
  • JIANG Lingyong
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  • 1.Department of Oral and Maxillofacial Surgery, Shanghai Ninth People′s Hospital, Shanghai Jiao Tong University School of Medicine; College of Stomatology, Shanghai Jiao Tong University; National Center for Stomatology; National Clinical Research Center for Oral Diseases; Shanghai Key Laboratory of Stomatology; Shanghai Research Institute of Stomatology, Shanghai 200011, China
    2.Department of Oral Maxillofacio-Head and Neck Oncology, Shanghai Ninth People′s Hospital, Shanghai Jiao Tong University School of Medicine; College of Stomatology, Shanghai Jiao Tong University; National Center for Stomatology; National Clinical Research Center for Oral Diseases; Shanghai Key Laboratory of Stomatology; Shanghai Research Institute of Stomatology, Shanghai 200011, China
    3.Department of Immunology and Microbiology, Shanghai Jiao Tong University School of Basic Medical Sciences; Shanghai Institute of Immunology, Shanghai 200025, China
    4.Department of 2nd Dental Centre, Shanghai Ninth People′s Hospital, Shanghai Jiao Tong University School of Medicine; College of Stomatology, Shanghai Jiao Tong University; National Center for Stomatology; National Clinical Research Center for Oral Diseases; Shanghai Key Laboratory of Stomatology; Shanghai Research Institute of Stomatology, Shanghai 200011, China
JANG Lingyong, E-mail: jianglingyong@sjtu.edu.cn.

Received date: 2025-02-19

  Accepted date: 2025-04-29

  Online published: 2025-09-30

Supported by

“Two-Hundred Talents” Program of Shanghai Jiao Tong University School of Medicine(20221809);National Natural Science Foundation of China(82430032);National Key Research and Development Program of China(2024YFC2510700);Shanghai Science and Technology Innovation Action Plan International Science and Technology Cooperation Program(23410713600);Natural Science Foundation of Shanghai(21ZR1436900);Cross-Disciplinary Research Fund of Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine(JYJC202116);Biomaterials and Regenerative Medicine Institute Cooperative Research Project, Shanghai Jiao Tong University School of Medicine(2022LHB02)

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摘要

目的·探究小鼠颌骨成骨细胞对B细胞分化发育的调控效应及具体作用机制。方法·采用优化的酶消化法制备小鼠颌骨单细胞悬液，体外诱导分化为成骨细胞。通过实时荧光定量聚合酶链反应（RT-qPCR）、碱性磷酸酶（ALP）染色及茜素红S（ARS）染色验证成骨细胞的成骨潜能。通过免疫荧光染色观察小鼠颌骨组织中成骨细胞与B细胞的空间定位关系。采用流式细胞分选技术获得高纯度、高活性的造血谱系前体细胞。通过建立基于Transwell小室的共培养体系，探究不同浓度颌骨成骨细胞（5×104、2.5×105、5×105个/孔）对B细胞分化（5×104个/孔）发育的调节作用。通过流式细胞术及RT-qPCR检测不同浓度成骨细胞诱导下B细胞的活性与分化程度。利用RT-qPCR检测成骨细胞成骨分化过程中与B细胞发育相关的外分泌因子的表达情况。结果·成骨诱导后，ALP与ARS染色结果显示小鼠颌骨成骨细胞具有优越的成骨潜能。RT-qPCR结果显示成骨细胞内高表达成骨基因Runx2、Osx、Ocn和Alp。组织免疫荧光检测结果显示，小鼠颌骨组织内成骨细胞与B细胞在空间定位上紧密相邻。在体外共培养模型中，流式细胞分析结果显示，成骨细胞以浓度依赖性的方式促进B细胞分化与发育。RT-qPCR与细胞免疫荧光检测结果显示，成骨细胞上调B细胞发育关键基因Ebf1、Rag1、Il7r和Pax5（均P<0.001）。RT-qPCR结果显示，与B细胞发育密切相关的细胞因子Il7、Baff和Flt3l在成骨细胞成骨分化的过程中显著上调（均P<0.05）。结论·颌骨成骨细胞以浓度依赖性的方式促进B细胞分化与发育，成骨细胞可能通过分泌生长因子上调B细胞内分化关键基因的表达。

关键词： 颌骨成骨细胞; 骨免疫; B细胞; 免疫调节

本文引用格式

王歆雨 , 陈芊烨 , 孙计萍 , 鲁婷玮 , 黄湘如 , 孙思远 , 刘媛琪 , 潘厚文 , 代庆刚 , 沈蕾 , 江凌勇 . 颌骨成骨细胞调控B细胞分化的效应研究[J]. 上海交通大学学报（医学版）, 2025 , 45(9) : 1106 -1115 . DOI: 10.3969/j.issn.1674-8115.2025.09.003

Abstract

Objective ·To investigate the regulatory effects and underlying mechanisms of mouse mandibular osteoblasts on B cell differentiation and development. Methods ·Single-cell suspensions from mouse mandibular bone were prepared using an optimized enzymatic digestion method and induced to differentiate into osteoblasts in vitro. Osteogenic potential was validated by real-time quantitative PCR (RT-qPCR), alkaline phosphatase (ALP) staining, and alizarin red S (ARS) staining. The spatial localization relationship between osteoblasts and B cells in mandibular tissues was examined via immunofluorescence staining. High-purity hematopoietic progenitor cells were isolated using fluorescence-activated cell sorting. A Transwell co-culture system was established to assess the regulatory effects of different osteoblast concentrations (5×104, 2.5×105, and 5×105 cells/well) on B cell differentiation (5×104 cells/well). Flow cytometry and RT-qPCR were employed to evaluate B cell viability and differentiation. Additionally, RT-qPCR was used to analyze the expression of osteoblast-secreted factors associated with B cell development during osteogenic differentiation. Results ·Mandibular osteoblasts exhibited robust osteogenic potential, as confirmed by ALP/ARS staining and high expression of osteogenic markers (Runx2, Osx, Ocn, and Alp) via RT-qPCR. Immunofluorescence revealed close spatial proximity between osteoblasts and B cells in mandibular tissues. In the co-culture system, osteoblasts promoted B cell differentiation in a concentration-dependent manner. RT-qPCR and immunofluorescence demonstrated that osteoblasts significantly upregulated key genes involved in B cell development (Ebf1, Rag1, Il7r, and Pax5; all P<0.001). Furthermore, osteoblast-derived factors (Il7, Baff, and Flt3l) were markedly elevated during osteogenic differentiation (all P<0.05). Conclusion ·Mandibular osteoblasts enhance B cell differentiation and development in a concentration-dependent manner, likely through secreting growth factors that upregulate critical B cell differentiation genes.

Key words： jaw osteoblasts; osteoimmunology; B lymphocytes; immunoregulation

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