PDGF-BB overexpressing dental pulp stem cells improve angiogenesis in dental pulp regeneration

Wentao Jiang ^1,2,3,4 Shuhan Duan ^2,3,4,5 Weiping Li ^6,7,8 HUIJIAO YAN ^2,3,4,9 Chenli Si ^10,3,4 Ningwei Xu ^1,2,3,4 Yishuai Li ^1,2,3,4 Wenjie Zhang ^2,3,4,9* Shensheng Gu ^1,2,3,4*

• ¹ Department of Endodontics, Shanghai Ninth People’s Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China
• ² College of Stomatology, School of Medicine, Shanghai Jiao Tong University, Pudong, Shanghai Municipality, China
• ³ National Center for Stomatology & National Clinical Research Center for Oral Diseases, Shanghai, China
• ⁴ Shanghai Key Laboratory of Stomatology & Shanghai Research Institute of Stomatology, Shanghai, China
• ⁵ Department of Oral Surgery, Shanghai Ninth People’s Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, Shanghai Municipality, China
• ⁶ Affiliated Stomatological Hospital, Nanjing Medical University, Nanjing, Jiangsu Province, China
• ⁷ State Key Laboratory Cultivation Base of Research, Prevention and Treatment for Oral Diseases, Nanjing, China
• ⁸ Jiangsu Province Engineering Research Center of Stomatological Translational Medicine, Nanjing, China
• ⁹ Department of Prosthodontics, Shanghai Ninth People’s Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, China
• ¹⁰ Department of Dermatology, Shanghai Ninth People’s Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, China

Introduction: 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. Methods: 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. Results and Discussion: 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. In vivo 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.