Effect of jaw osteoblasts on B cell development via cytokine secretion
10.3969/j.issn.1674-8115.2025.09.003
- VernacularTitle:颌骨成骨细胞调控B细胞分化的效应研究
- Author:
Xinyu WANG
1
;
Qianye CHEN
;
Jiping SUN
;
Tingwei LU
;
Xiangru HUANG
;
Siyuan SUN
;
Yuanqi LIU
;
Houwen PAN
;
Qinggang DAI
;
Lei SHEN
;
Lingyong JIANG
Author Information
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
- Publication Type:Journal Article
- Keywords:
jaw osteoblasts;
osteoimmunology;
B lymphocytes;
immunoregulation
- From:
Journal of Shanghai Jiaotong University(Medical Science)
2025;45(9):1106-1115
- CountryChina
- Language:Chinese
-
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.
