Development and Validation of Large-Sized Engineered Cartilage Constructs in Full –Thickness Chondral Defects in a Rabbit Model
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Long-term applicability of current surgical interventions for the repair of articular cartilage is jeopardized by the formation of mechanically inferior repair tissue. Cartilage tissue engineering offers the possibility of developing functional repair tissue, similar to that of native cartilage, enabling long-lasting repair of cartilage defects. Current techniques, however, rely on the need for a large number of cells, requiring substantial harvesting of donor tissue or a separate cell expansion phase. As routine cell expansion methods tend to elicit negative effects on cell function, the following study describes an approach to generate large-sized engineered cartilage constructs (≥ 3 cm2) directly from a small number of immature rabbit chondrocytes (approximately 20,000), without the use of a scaffold. After characterizing the hyaline-like engineered constructs, the in vivo repair capacity was assessed in a chondral defect model in the patellar groove of rabbits. In vitro remodeling of the constructs developed in the bioreactor occurred as early as 3 weeks, with the histological staining exhibiting zonal differences throughout the depth of the tissue. With culturing parameters optimized (3 weeks growth under 15 mM NaHCO3), constructs were grown and implanted into critical-sized 4 mm chondral defects. Assessed after 1, 3 and 6 months (n=6), implants were scored macroscopically to evaluate integration and survival of the implants. Out of 18 rabbits, 16 received normal or nearly normal over-all repair assessment. Histological and immunohistochemical evaluation showed good integration with surrounding cartilage and underlying subchondral bone. Architectural remodeling of the constructs was present at each time point, with the presence of flattened chondrocytes at the implant surface and columnar arrangement of chondrocytes in deeper zones. The observation of in vivo remodeling was also supported by the changes in biochemical composition of the constructs. At each time point, constructs had a collagen to proteoglycan ratio similar to that of native cartilage (3:1 collagen to proteoglycan). In contrast, the repair tissue for each control group was inferior to that produced with treated defects. These initial results hold promise for the generation of engineered articular cartilage for the clinical repair of cartilage defects without the limitations of current surgical repair strategies.