Jiang Wentao, Duan Shuhan, Li Weiping, Yan Huijiao, Si Chenli, Xu Ningwei, Li Yishuai, Zhang Wenjie, Gu Shensheng
Department of Endodontics, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
College of Stomatology, Shanghai Jiao Tong University, Shanghai, China.
Front Bioeng Biotechnol. 2025 Apr 24;13:1578410. doi: 10.3389/fbioe.2025.1578410. eCollection 2025.
Angiogenesis represents a critical challenge in dental pulp regeneration due to the tissue's restricted nutrient supply through a 0.5-mm apical foramen. While dental pulp stem cells (DPSCs) hold regenerative potential, their limited vascularization capacity impedes clinical applications. Through Single-cell RNA sequencing (scRNA-seq) analysis of human dental pulp, we discovered a PDGF (+) mesenchymal subset exhibiting enhanced angiogenic signatures, suggesting targeted cell selection could overcome this bottleneck.
ScRNA-seq identified PDGF (+) subpopulation in human pulp samples, validated through multiplex immunohistochemical of the localization of PDGF/CD73/CD31. PDGF-BB-overexpressing DPSCs were engineered via lentiviral vectors. Functional assessments included: 1) CCK-8/Edu/cell cycle/transwell assays for proliferation and migration ability 2) HUVECs co-culture models analyzing chemotaxis and tube formation 3) Vascularized tissue formation in rat kidney capsule transplants.
The CD73 (+) PDGF (+) subpopulation demonstrated spatial correlation with CD31 (+) vasculature. PDGF-BB overexpression enhanced DPSCs' proliferative capacity and migration capacity. Co-cultured HUVECs exhibited increased tube formation with PDGF-BB group. transplants generated more vascular structures containing CD31 (+) endothelia. These findings establish PDGF-BB engineering as an effective strategy to amplify DPSCs' angiogenic potential, while emphasizing the therapeutic value of functionally-defined stem cell subpopulations in pulp regeneration.
由于牙髓组织通过0.5毫米的根尖孔获得的营养供应有限，血管生成是牙髓再生中的一项关键挑战。虽然牙髓干细胞（DPSCs）具有再生潜力，但其有限的血管生成能力阻碍了临床应用。通过对人牙髓进行单细胞RNA测序（scRNA-seq）分析，我们发现了一个表现出增强血管生成特征的血小板衍生生长因子（PDGF）阳性间充质亚群，这表明靶向细胞选择可以克服这一瓶颈。
scRNA-seq在人牙髓样本中鉴定出PDGF阳性亚群，并通过PDGF/CD73/CD31定位的多重免疫组织化学进行验证。通过慢病毒载体构建过表达PDGF-BB的DPSCs。功能评估包括：1）CCK-8/Edu/细胞周期/Transwell实验检测增殖和迁移能力；2）人脐静脉内皮细胞（HUVECs）共培养模型分析趋化性和管形成；3）大鼠肾囊移植中的血管化组织形成。
CD73阳性PDGF阳性亚群与CD31阳性脉管系统存在空间相关性。PDGF-BB过表达增强了DPSCs的增殖能力和迁移能力。与PDGF-BB组共培养的HUVECs管形成增加。移植产生了更多含有CD31阳性内皮的血管结构。这些发现确立了PDGF-BB工程作为一种放大DPSCs血管生成潜力的有效策略，同时强调了功能定义的干细胞亚群在牙髓再生中的治疗价值。
