The role of chondrocyte senescence in the pathogenesis of osteoarthritis and in limiting cartilage repair.

BACKGROUND

With increasing age, the prevalence of osteoarthritis increases and the efficacy of articular cartilage repair decreases. As chondrocytes age, they synthesize smaller, less uniform aggrecan molecules and less functional link proteins, their mitotic and synthetic activity decline, and their responsiveness to anabolic mechanical stimuli and growth factors decreases. These observations led us to hypothesize that progressive cell senescence decreases the ability of chondrocytes to maintain and to restore articular cartilage.

METHODS

To test this hypothesis, we measured cell senescence markers (beta-galactosidase expression, mitotic activity, and telomere length) in human articular cartilage chondrocytes from twenty-seven donors ranging in age from one to eighty-seven years. We also assessed mitochondrial DNA, membrane potential, and numerical density. To determine if chondrocyte age changes are reversible, we transfected human articular cartilage chondrocytes with the human telomerase gene (hTERT) and human papilloma virus oncogenes (E6 and E7).

RESULTS

Beta-galactosidase expression increased with age (r = 0.84, p = 0.0001), while mitotic activity and telomere length declined (r = -0.77, p = 0.001 and r = -0.71, p = 0.0004, respectively). Decreasing telomere length was closely correlated with increasing expression of beta-galactosidase and decreasing mitotic activity. As the number of population doublings increased, mitochondrial DNA was degraded, mitochondrial membrane potential was lost, and the number of mitochondria per cell declined. Transfection of human articular cartilage chondrocytes from a forty-seven-year-old donor with hTERT and human papilloma virus proto-oncogenes E6 and E7 created a cell line that has completed more than 300 population doublings as compared with an upper limit of twenty-five population doublings for normal cells. Telomere length increased in cells transduced with hTERT.

CONCLUSIONS

These findings help to explain the previously reported age-related declines in chondrocyte synthetic activity, mitotic activity, and responsiveness to anabolic cytokines and mechanical stimuli. They also suggest that in vivo chondrocyte senescence contributes to the age-related increase in the prevalence of osteoarthritis and decrease in the efficacy of cartilage repair. The creation of immortal cells with increased telomere length suggests that the progression of human chondrocytes toward senescence is not inevitable.