In VivoMaturation of Scaffold-free Engineered Articular Cartilage on Hydroxyapatite

Abstract

Tissue engineering is a promising approach, not only for cartilage, but also for osteochondral repair. Recent studies have demonstrated that scaffold-free cartilaginous tissue can be engineered using the alginate-recovered-chondrocyte (ARC) method. This method has also been applied to form osteochondral tissue using bovine articular chondrocytes and coralline hydroxyapatite (HA). The purpose of this study was to test whether osteochondral tissue, fabricated in vitro using the ARC method combined with a block of HA, would undergo maturation in vivo using a subcutaneous model in immunodeficient mice. Articular chondrocytes were isolated from the cartilage of New Zealand white rabbits and cultured in alginate beads. The cells with their associated matrix were recovered by dissolving the alginate beads with a sodium citrate buffer, resuspended in media and seeded onto a porous HA block. After 4 weeks of culture, some samples were analyzed, and others were implanted into subcutaneous pockets in nude mice. The analysis involved removing the cartilage portion of the de novo-formed ARC-HA graft and performing biochemical and histological examinations. Some samples were subjected to nondecalcified histology. Histological and immunohistochemical analyses of cartilaginous tissue were performed at 0, 2, 4, and 8 weeks after implantation. Biochemical characteristics were examined at 0, 4, and 8 weeks. The size and shape of the implanted ARC osteochondral tissue changed with time. The histological and immunohistochemical examination of the tissue revealed that it contained a cartilage-like matrix that stained strongly with Toluidine blue and for collagen type II. The proteoglycan (PG) content had increased significantly at 4 weeks from baseline. However, by 8 weeks, the PG content had decreased from 4 weeks. The results presented here represent a possible approach to form a tissue-engineered osteochondral implant. Further studies are needed to improve biomechanical properties of the osteochondral implant to be suitable for surgical transplantation.