Neovascularization and free microsurgical transfer of in vitro cartilage-engineered constructs

Cartilage tissue engineering shows to have tremendous potential for the reconstruction of three-dimensional cartilage defects. To ensure survival, shape, and function, in vitro cartilage-engineered constructs must be revascularized. This article presents an effective method for neovascularization and free microsurgical transfer of these in vitro constructs. Twelve female Chinchilla Bastard rabbits were used. Cartilage-engineered constructs were created by isolating chondrocytes from auricular biopsies, amplifying in monolayer culture, and then seeding them onto polycaprolactone scaffolds. In each prefabricated skin flap, three in vitro cartilage-engineered constructs (2 x 2 x 0.5 cm) and one construct without cells (served as the control) were implanted beneath an 8 x 15 cm random-pattern skin flap, neovascularized by implantation of an arteriovenous vascular pedicle with maximal blood flow. Six weeks later, the neovascularized flaps with embedded cartilage-engineered constructs were completely removed based on the newly implanted vascular pedicle, and then freely retransferred into position using microsurgery. Macroscopic observation, selective microangiography, histology, and immunohistochemistry were performed to determine the construct vitality, neovascularization, and new cartilage formation. The results showed that all neovascularized skin flaps with embedded constructs were successfully free-transferred as free flaps. The implanted constructs were well integrated and protected within the flap. All constructs were well neovascularized and showed histologically stability in both size and form. Immunohistology showed the existence of cartilage-like tissue with extracellular matrix neosynthesis.