Chondrocyte apoptosis and nitric oxide production during experimentally induced osteoarthritis.
OBJECTIVE
Chondrocytes produce nitric oxide (NO) and undergo apoptosis in response to exogenous NO. This study sought to examine the relationship between NO synthesis, chondrocyte apoptosis, and the development of cartilage degradation during experimental osteoarthritis (OA).


METHODS
OA was induced in rabbits by anterior cruciate ligament transection (ACLT). Knees were harvested after 4 weeks and assessed for OA severity and chondrocyte apoptosis. Conditioned media from cultured cartilage explants were analyzed for nitrite content. Cartilage sections were analyzed by immunohistochemistry for the presence of nitrotyrosine.


RESULTS
All ACLT knees demonstrated osteoarthritic changes. Conditioned media from ACLT cartilage organ cultures contained higher levels of nitrite as compared with cartilage samples from the nonoperated side or from rabbits that had not received ACLT. Cultures of specific areas of cartilage from ACLT knees showed high levels of NO production in the medial femoral and medial tibial cartilage. Approximately 28.7% of chondrocytes isolated from ACLT cartilage and 6.7% of chondrocytes from cartilage of the nonoperated side underwent apoptosis. In situ staining demonstrated apoptotic cells in the superficial and middle zones of ACLT cartilage. A high number of apoptotic cells was present at the pannus-cartilage junction. In control cartilage, the superficial zone contained a small number of cells in apoptosis. The prevalence of apoptotic cells was significantly correlated with the levels of nitrite production and OA grade.


CONCLUSIONS
These observations suggest that, during the early phases of OA, NO production may lead to chondrocyte apoptosis, and that both events contribute to the pathogenesis of cartilage degradation. Inhibitors of NO synthesis and chondrocyte apoptosis may therefore be of therapeutic value after cartilage injury and in patients with OA.
Hashimoto
S
S
The Scripps Research Institute, La Jolla, California 92937, USA.
Takahashi
K
K
Amiel
D
D
Coutts
R D
RD
Lotz
M
M
D000818:Animals; D017209:Apoptosis; D002358:Cartilage, Articular / Q000378:metabolism / Q000473:pathology*; D019902:Chondrocytes / Q000378:metabolism / Q000473:pathology*; D017077:Culture Media, Conditioned / Q000378:metabolism; D046508:Culture Techniques; D005434:Flow Cytometry; D007124:Immunoenzyme Techniques; D009569:Nitric Oxide / Q000378:metabolism*; D010003:Osteoarthritis / Q000473:pathology*; D011509:Proteoglycans / Q000378:metabolism; D011817:Rabbits; D014443:Tyrosine / Q000008:administration & dosage / Q000378:metabolism
D017077:Culture Media, Conditioned; D011509:Proteoglycans; D009569:Nitric Oxide; D014443:Tyrosine