Wang Xinyu, Chen Qianye, Sun Jiping, Huang Zihan, Zhang Zijian, Lu Tingwei, Huang Xiangru, Sun Siyuan, Liu Yuanqi, Pan Houwen, Mei Li, Cooper Paul R, Dai Qinggang, Shen Lei, Jiang Lingyong
Center of Craniofacial Orthodontics, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai Research Institute of Stomatology, Shanghai, China.
Department of Oral and Maxillofacial-Head and Neck Oncology, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
Front Bioeng Biotechnol. 2025 Aug 11;13:1596143. doi: 10.3389/fbioe.2025.1596143. eCollection 2025.
The bone marrow microenvironment, comprising various cell types and molecular signals, finely orchestrates the self-renewal and lineage commitment of hematopoietic stem cells (HSCs). Although most investigations have centered on mesenchymal stem cells (MSCs) from long bones, the distinct properties and immunoregulatory functions of craniofacial bone marrow derived MSCs remain largely unexplored. Notably, jawbone MSCs not only exhibit a robust capacity for promoting hematopoietic regeneration but also offer therapeutic potential in infectious diseases.
Using an optimized enzymatic digestion protocol, we obtained a highly viable single-cell suspension from mouse jawbone . Single-cell sequencing was then performed to explore the interactions between jawbone MSCs and HSCs, while tissue immunofluorescence clarified their spatial distribution. osteogenic and adipogenic differentiation assays confirmed the multilineage potential of jawbone MSCs. A biomimetic co-culture system, designed to emulate the bone marrow niche, was employed to assess the impact of jawbone MSCs on HSC differentiation, which was evaluated via flow cytometry. Mechanistic insights into HSC changes were gleaned from RT-qPCR and cellular immunofluorescence. Subsequently, an LPS-induced acute infection model was established to evaluate the therapeutic efficacy of jawbone MSCs. Finally, comprehensive analysis of single-cell sequencing data, in conjunction with RT-qPCR findings, elucidated the regulatory pathways through which jawbone MSCs promote hematopoiesis.
Single-cell sequencing revealed a robust interaction between jawbone MSCs and HSCs. Tissue immunofluorescence demonstrated that in the mouse jawbone, MSCs and HSCs were located in close spatial proximity. osteogenic and adipogenic induction experiments showed that jawbone MSCs possess considerable multilineage differentiation potential. Co-culture assays further indicated that jawbone MSCs induce HSCs to differentiate into various immune cell types, particularly promoting B cell generation. RT-qPCR and immunofluorescence assays confirmed that pivotal transcription factors, such as PAX5, were activated in B cells. In an infection model, jawbone MSCs exhibited significant anti-infective capabilities, effectively reducing mortality and systemic inflammation in infected mice. A deeper analysis of the single-cell sequencing data revealed that jawbone MSCs mainly facilitate hematopoiesis by secreting CXCL12.
Through single-cell sequencing, multilineage induction, co-culture systems, and a mouse model of LPS-induced acute infection, this study systematically elucidates the close interplay between jawbone MSCs and HSCs, as well as their pivotal roles in immune modulation and anti-infective responses. The findings demonstrate that jawbone MSCs not only exhibit robust multilineage differentiation potential but also secrete CXCL12 and activate key B cell transcription factors (such as PAX5). This process significantly promotes HSC differentiation into B cells, improves survival rates in infected mice, and attenuates systemic inflammation. These results establish a strong foundation for further investigation into the applications of jawbone MSCs in immune regulation and disease therapy.
骨髓微环境由多种细胞类型和分子信号组成，精细地调控着造血干细胞（HSC）的自我更新和谱系定向。尽管大多数研究集中于长骨来源的间充质干细胞（MSC），但颅面骨骨髓来源的MSC的独特特性和免疫调节功能在很大程度上仍未被探索。值得注意的是，颌骨MSC不仅具有促进造血再生的强大能力，还在传染病治疗中具有潜在应用价值。
采用优化的酶消化方案，我们从小鼠颌骨获得了高活力的单细胞悬液。随后进行单细胞测序以探索颌骨MSC与HSC之间的相互作用，同时组织免疫荧光明确了它们的空间分布。成骨和成脂分化实验证实了颌骨MSC的多谱系分化潜能。设计了一种仿生共培养系统来模拟骨髓微环境，用于评估颌骨MSC对HSC分化的影响，通过流式细胞术进行评估。从RT-qPCR和细胞免疫荧光中获取了关于HSC变化的机制性见解。随后，建立了脂多糖诱导的急性感染模型以评估颌骨MSC的治疗效果。最后，结合RT-qPCR结果对单细胞测序数据进行综合分析，阐明了颌骨MSC促进造血的调控途径。
单细胞测序揭示了颌骨MSC与HSC之间的强大相互作用。组织免疫荧光表明，在小鼠颌骨中，MSC和HSC在空间上紧密相邻。成骨和成脂诱导实验表明颌骨MSC具有相当大的多谱系分化潜能。共培养实验进一步表明，颌骨MSC诱导HSC分化为多种免疫细胞类型，尤其促进B细胞生成。RT-qPCR和免疫荧光实验证实，关键转录因子如PAX5在B细胞中被激活。在感染模型中，颌骨MSC表现出显著的抗感染能力，有效降低了感染小鼠的死亡率和全身炎症反应。对单细胞测序数据的深入分析表明，颌骨MSC主要通过分泌CXCL12促进造血。
通过单细胞测序、多谱系诱导、共培养系统以及脂多糖诱导的急性感染小鼠模型，本研究系统地阐明了颌骨MSC与HSC之间的密切相互作用，以及它们在免疫调节和抗感染反应中的关键作用。研究结果表明，颌骨MSC不仅具有强大的多谱系分化潜能，还分泌CXCL12并激活关键的B细胞转录因子（如PAX5）。这一过程显著促进HSC分化为B细胞，提高感染小鼠的存活率，并减轻全身炎症。这些结果为进一步研究颌骨MSC在免疫调节和疾病治疗中的应用奠定了坚实基础。
