颌骨成骨细胞调控B细胞分化的效应研究
Effect of jaw osteoblasts on B cell development via cytokine secretion
摘要
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.关键词
颌骨成骨细胞/骨免疫/B细胞/免疫调节Key words
jaw osteoblasts/osteoimmunology/B lymphocytes/immunoregulation分类
医药卫生引用本文复制引用
王歆雨,沈蕾,江凌勇,陈芊烨,孙计萍,鲁婷玮,黄湘如,孙思远,刘媛琪,潘厚文,代庆刚..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):1106-1115,10.基金项目
上海交通大学医学院"双百人"项目(20221809) （20221809）
国家自然科学基金(82430032,81870740,82071083,82271006) （82430032,81870740,82071083,82271006）
国家重点研发计划(2024YFC2510700) （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) （23410713600）
上海市自然科学基金(21ZR1436900,22ZR1436700) （21ZR1436900,22ZR1436700）
上海交通大学医学院附属第九人民医院交叉研究基金(JYJC202116) （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). "Two-Hundred Talents"Program of Shanghai Jiao Tong University School of Medicine(20221809) （2022LHB02）
National Natural Science Foundation of China(82430032,81870740,82071083,82271006) （82430032,81870740,82071083,82271006）
National Key Research and Development Program of China(2024YFC2510700) （2024YFC2510700）
Shanghai Science and Technology Innovation Action Plan International Science and Technology Cooperation Program(23410713600) （23410713600）
Natural Science Foundation of Shanghai(21ZR1436900,22ZR1436700) （21ZR1436900,22ZR1436700）
Cross-Disciplinary Research Fund of Shanghai Ninth People's Hospital,Shanghai Jiao Tong University School of Medicine(JYJC202116) （JYJC202116）
Biomaterials and Regenerative Medicine Institute Cooperative Research Project,Shanghai Jiao Tong University School of Medicine(2022LHB02). （2022LHB02）
