Bi-layered Composite Scaffold for Repair of the Osteochondral Defects.

Osteochondral defect presents a big challenge for clinical treatment. This study aimed at constructing a bi-layered composite chitosan/chitosan-β-tricalcium phosphate (CS/CS-β-TCP) scaffold and at repairing the rat osteochondral defect. The bi-layered CS/CS-β-TCP scaffold was fabricated by lyophilization, and its microstructure was observed by a scanning electron microscope. Chondrocytes and bone marrow stem cells (BMSCs) were seeded into the CS layer and the CS-β-TCP layer, respectively. Viability and proliferation ability of the cells were observed under a confocal microscope. After subcutaneous implantation, the chondrogenic ability of the CS layer and osteogenic ability of the CS-β-TCP layer were evaluated by immunofluorescence. Then, the bi-layered scaffolds were implanted into the rat osteochondral defects and the harvested samples were macroscopically and histologically evaluated. The bi-layered CS/CS-β-TCP scaffold exhibited the distinctive microstructures for each layer. The seeded chondrocytes in the CS layer could maintain the chondrogenic lineage, whereas BMSCs in the CS-β-TCP layer could continually differentiate into the osteogenic lineage. Moreover, cells in both layers could maintain well viability and excellent proliferation ability. For the study, the newly formed tissues in the bi-layered scaffolds group were similar with the native osteochondral tissues, which comprised hyaline-like cartilage and subchondral bone, with better repair effects compared with those of the pure CS group and the blank control group. This is the first time that the bi-layered composite CS/CS-β-TCP scaffold has been fabricated and evaluated with respect to osteochondral defect repair. The bi-layered CS/CS-β-TCP scaffolds could facilitate osteochondral defect repair and might be the promising candidates for osteochondral tissue engineering.