In Vivo Bone Regeneration Evaluation

All animal experiments were conducted under the ethical committee guidelines of the Affiliated Stomatological Hospital of Chongqing Medical University. Cells from each group (approximately 3×10⁶ cells per group) were loaded on β-tricalcium phosphate (β-TCP) porcelain granules (Bio-lu Biomaterials, Shanghai, China) and subcutaneously implanted the mixture into the flanks of 6-week-old BALB/c nude mice. 8 weeks later, these implants were obtained, fixed with paraformaldehyde (4%), decalcified in EDTA decalcification solution (Servicebio, Wuhan, China), and embedded in paraffin. Tissues embedded were serially sliced (5 μm) and processed for hematoxylin and eosin (H&E; Solarbio, Beijing, China) staining and Masson trichrome (Solarbio, Beijing, China) staining. Furthermore, the immunohistochemistry (IHC) staining was also carried out with anti-OCN antibodies (Abcam, Cambridge, UK) as previously described (33 (link)). The images were acquired by digital section scanner VS200 (Olympus, Japan).