Phenotype-Preserving Co-culture of Osteoblasts and Chondrocytes Enhances Bone-Cartilage Interface Integration in a PRP-Augmented Scaffold

表型保持的成骨细胞和软骨细胞共培养可增强富血小板血浆增强支架中的骨-软骨界面整合

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Abstract

BACKGROUND: Effective bone-cartilage integration remains a challenge in orthopedic surgery. Conventional methods often fail to reconstruct the native osteochondral interface. This study explores a scaffold-mediated approach utilizing co-cultured osteoblasts and chondrocytes, with platelet-rich plasma (PRP) as a potential promotor for bone-cartilage interface healing. METHODS: We developed a co-culture system integrating both osteoblasts and chondrocytes on PLGA scaffolds, either with or without PRP supplementation. Cell phenotype maintenance was evaluated by RT-PCR, while morphological analysis was performed by scanning electron microscopy and fluorescence microscopy. To assess healing potential, we created a gap-mimic construct comprising bone, scaffold, and cartilage layers, which was implanted subcutaneously in BALB/c-nude mice. Gap healing was evaluated at 4 and 8 weeks through macroscopic examination, quantitative adhesion analysis, and histological assessment of cellular invasion. RESULTS: Co-cultured osteoblasts and chondrocytes maintained their phenotypes on PLGA scaffolds, with PRP significantly enhancing cell adhesion (215% increase for chondrocytes, 120% for osteoblasts) and proliferation. In vivo, cell-containing scaffolds demonstrated significantly greater attachment at the bone-cartilage interface compared to acellular constructs. PRP-treated scaffolds exhibited higher attachment rates (82.3% vs 76.7%) and cellular invasion (5/6 vs 3/6 constructs) at 8 weeks, with invasion observed as early as 4 weeks in the PRP group, suggesting accelerated remodeling. CONCLUSION: This study demonstrates the feasibility of developing transplantable scaffolds containing co-cultured osteoblasts and chondrocytes while preserving their phenotypes. These scaffolds exhibit significant potential in promoting healing at the bone-cartilage interface, with PRP further enhancing proliferation and improving the scaffold's ability to promote bone-cartilage interface healing.

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