Successful bone regeneration after dental and craniofacial reconstruction can present a significant clinical challenge. In this study, we investigated whether the regenerative potential of human periosteum-derived stromal progenitor cells (PDCs) depends on their anatomical origin. Paired periosteum samples were collected from jaw and appendicular bones (fibula, femur, or iliac crest) of five donors and expanded in a xeno-free, platelet lysate-supplemented medium. All cultures exhibited a typical fibroblastic morphology and a uniform stromal immunophenotype (≥95% CD73/90/105 and ≤2% CD45/14/19/HLA-DR). PDCs were tested in a 3D cartilage disc formation assay after seeding on collagen-coated transwell inserts or on transferrable porcine collagen membranes. Appendicular-derived PDCs consistently formed stable, cartilage-like 3D discs with a mean weight of 14.7 mg and a range of 8.8-23.6 mg, whereas jaw-derived PDCs produced contracted, irregular aggregates. Histological and molecular analyses confirmed greater proteoglycan deposition and higher COL2A1 expression in appendicular-derived PDCs, indicating superior intrinsic chondrogenic potential. Culturing jaw-derived PDCs on porcine collagen membranes did not enhance their differentiation capacity. Our findings demonstrate that the chondrogenic potential of PDCs is strongly determined by their anatomical origin. Therefore, selecting the appropriate periosteal source, combined with optimized biomaterials, is crucial for achieving predictable and efficient bone regeneration.
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