Proteoglycan synthesis in chondrocyte cultures from osteoarthrotic and normal human articular cartilage

Chondrocytes derived by outgrowth from normal and osteoarthrotic human femoral head cartilage were grown in high density cultures for five passages. Cultures were analysed for their sulfated macromolecular components in medium, layer-matrix and intracellular compartments. Two fractions were obtained: typical proteoglycans and glycosaminoglycan-peptides (Mr approx. 60 000) which might result from an enzymatic degradation of proteoglycans. Proteoglycans from normal and osteoarthrotic cultures exhibited similar biochemical properties (size, protein:uronic acid ratio, glycosaminoglycan composition). Slightly less proteoglycans were aggregated with hyaluronic acid in osteoarthrotic than in normal cultures. Three populations of proteoglycan subunit were obtained under dissociative conditions (Sepharose CL-2B) in both normal and osteoarthrotic cultures: proteoglycan 1 (Kav=0.04), proteoglycan 2 (Kav=0.26) which were aggregated with hyaluronic acid in associative conditions, and proteoglycan 4 (Kav=0.48). A fourth population, proteoglycan 3 (Kav=0.33, Sepharose 2B and CL-2B) was intracellular in osteoarthrotic cultures. After a 4 day incubation period, about 60% more proteoglycans were found in osteoarthrotic than in normal cultures (medium+52%, layer-matrix + 44% and 17 times the normal value inside the cells). Proteoglycan distribution kinetics in the three compartments showed a higher net accumulation of proteoglycans in osteoarthrotic-derived cultures.