Chondrocytes in culture produce a mechanically functional tissue

Abstract

A mechanically testable tissue was grown in vitro from rabbit chondrocytes that were initially plated at high density (approximately 80,000 cells/cm2). The DNA, collagen, and proteoglycan content, as well as the tissue thickness, tensile stiffness, and synthesis rates, were measured at 4, 6, and 8 weeks. The biochemical properties were similar to those for immature cartilage, with predominantly type-II collagen produced; this indicated that the cells retained their chondrocytic phenotype. The tissue formed a coherent mechanical layer with testable tensile stiffness as early as 4 weeks. The tensile elastic modulus reached 1.3 MPa at 8 weeks, which is in the range of values for native cartilage from the midzone. Collagen density was approximately 24 mg/ml at 8 weeks, which is about one-half the value for native cartilage, and the collagen fibril diameters were smaller. Chondrocytes in culture responded to culture conditions and were stimulated by cytokine interleukin-1β. When culture conditions were varied to RPMI nutrient medium with lower fetal bovine serum and higher ascorbic acid concentrations, the thickness decreased and the modulus increased significantly. Interleukin-1β, added to the 8-week culture for 2 weeks, caused a decrease of 60% in thickness, a decrease of 81% in proteoglycan content, and a decrease of 31% in collagen content; this is similar to the response of cartilage explants to interleukin-1β. This cartilage analog may be useful as a model system to study structure-function relationships in cartilage or as cartilage-replacement tissue.