TISSUE-ENGINEERED OSTEOCHONDRAL CONSTRUCTS IN THE SHAPE OF AN ARTICULAR CONDYLE

Abstract

Background: An entire articular condyle engineered from stem cells may provide an alternative therapeutic approach to total joint replacement. This study describes our continuing effort to optimize the chondrogenic and osteogenic differentiation from mesenchymal stem cells toward engineering articular condyles in vivo. Methods: Primary rat bone-marrow mesenchymal stem cells were induced to differentiate into chondrogenic and osteogenic lineages in vitro and were suspended in polyethylene glycol-based hydrogel. The hydrogel cell suspensions, each at a density of 20 × 106 cells/mL, were stratified into two separate layers that were molded into the shape and dimensions of an adult human cadaveric mandibular condyle by sequential photopolymerization. The osteochondral constructs fabricated in vitro were implanted in the dorsum of immunodeficient mice for twelve weeks. Results: De novo formation of articular condyles in the shape and dimensions of the adult human mandibular condyle occurred after a twelve-week period of in vivo implantation. Histological evaluation demonstrated two stratified layers of cartilaginous and osseous tissues, and yet there was mutual infiltration of cartilage-like and bone-like tissues into each other's territories. The cartilaginous portion was stained intensively to safranin O and expressed immunolocalized type-II collagen. Chondrocytes adjacent to the tissue-engineered osteochondral junction were enlarged and expressed type-X collagen, typical of hypertrophic chondrocytes. The osseous portion contained bone trabeculae-like structures and expressed immunolocalized type-I collagen, osteopontin, and osteonectin. Conclusions: A cell encapsulation density of 20 million cells/mL with in vivo incubation for twelve weeks yields further tissue maturation and phenotypic growth of both cartilage-like and bone-like tissues in the tissue-engineered articular condyle. Clinical Relevance: Tissue engineering of an entire condyle with chondral and osseous components derived from a single population of adult stem cells, as described in the present study, may have therapeutic implications in total joint replacement.