Subcutaneous Regeneration of Engineered Cartilage: A Comparison of Cell Sheets and Chondrocyte-Scaffold Constructs in a Porcine Model

Background: Stable cartilage regeneration in immunocompetent large animals remains a bottleneck problem that restricts clinical application. The inflammation elicited by degradation products of scaffolds has a decisive influence on cartilage formation. Although prolonged preculture in vitro could form mature engineered cartilage and allow sufficient degradation of scaffolds, the inflammatory reaction was still observed. This study explored the feasibility of using chondrocyte sheet technology to regenerate stable cartilage in the subcutaneous environment with a pig model. Methods: Passage 1 chondrocytes were used to form cell sheets by high-density culture. As a control, chondrocytes were seeded onto polyglycolic acid/polylactic acid scaffolds for 6 and 12 weeks' in vitro preculture, respectively. Then, they were autologously implanted subcutaneously into pigs for 2, 8, and 24 weeks. Gross view, histologic staining, and biochemical and biomechanical characteristics were evaluated. Results: With prolonged culture in vitro, relatively homogeneous engineered cartilages were formed with less scaffold residue. However, the chondrocyte-polyglycolic acid/polylactic acid group still encountered severe inflammation and inferior cartilage formation at 2 and 8 weeks in vivo. The engineered cartilage with cell sheet technique exhibited a relatively more stable and mature tissue structure without obvious inflammatory response at 24 weeks in vivo, which was similar to the native auricular cartilage. Conclusions: The chondrocyte sheet technique could successfully regenerate mature and stable engineered cartilages in pig models. It is possibly an effective method of repairing cartilage defects in the clinic that uses regenerated substitutes derived from autologous cell sheets.