Behavior of tissue-engineered human cartilage after transplantation into nude mice.
Cartilage lacks the ability to regenerate structural defects. Therefore, autologous grafting has been used routinely to replace cartilaginous lesions. Because tissue engineering of human cartilage with the help of bioresorbable polymer scaffolds is possible in experimental models, the demand for the clinical application grows. In this study we present an analysis of the behavior of transplants made of human chondrocyte pools, agarose and the resorbable polymer scaffold Ethisorb and a preliminary comparison with transplants made of single patients' cells and Ethisorb but without the additional ingredient agarose. Chondrocytes were isolated from the matrix of human septal cartilage by enzymatic digestion. The pool cells were kept in monolayer culture for 2 weeks, the single patients' cells for 3-4 weeks. Chondrocyte pools were suspended in agarose and seeded into the resorbable polymer scaffold Ethisorb. Single patients' cells were seeded without agarose. All cell-polymer constructs were kept in perfusion culture for 10-14 days and transplanted subcutaneously into thymusaplastic nude mice. Additionally we implanted Ethisorb fleeces embedded in agarose without chondrocytes. After 6, 12 and 24 weeks the animals were sacrificed and the specimens were explanted and analyzed histochemically and immunohistochemically. Polymer scaffolds not seeded with chondrocytes did not show cartilage formation. Resorption was complete after 12 weeks in vivo. Transplants from cell pools remained mechanically stable over 24 weeks apart from four transplants that were resorbed completely. Cartilage formation was observed in all pool-specimens with the presence of chondronic structures and a homogeneous matrix containing hyaline cartilage-specific matrix molecules such as collagen type II. Single patients' transplants showed hyaline cartilage matrix synthesis and mechanical stability as well. Chondrocyte pools are a suitable method to study cartilage engineering of human cells in vitro and in vivo in experimental models. Under clinical conditions it is, however, necessary to study the generation of cartilage from single patients' cells. We showed that it is possible without additional ingredients such as agarose. However, variations in the preliminary results show that the clinical application with human cells is more difficult than one would expect when using human chondrocyte pools. Further studies need to be performed to find out which individual factors influence the in vitro engineered cartilage's fate in vivo.