Chondrocyte death during murine osteoarthritis.
OBJECTIVE
To determine whether chondrocyte apoptosis occurs during the progression of osteoarthritis (OA) in the STR/ort mouse model of OA.


METHODS
Serial cryostat sections were cut (10 microns) through the knee joint of young and old male STR/ort mice and graded for the severity of OA lesions. Age- and sex-matched CBA mice were used as controls. Apoptotic chondrocytes were detected using the TUNEL assay. Ultrastructural changes were examined using electron microscopy (EM). Expression of biochemical markers associated with apoptosis (bax, bcl-2 and caspases-3, -8 & -9) was investigated using immunohistochemistry.


RESULTS
TUNEL assays on histological sections of STR/ort knee joints showed that the number of TUNEL-positive chondrocytes in the tibial medial articular cartilage correlated with the severity of the OA damage. These cells were located close to the lesional area. Only very occasional TUNEL positive chondrocytes were detected in either morphologically normal STR/ort cartilage or in control CBA cartilage. Ultrastructural analysis of chondrocytes neighboring focal osteoarthritic lesions in STR/ort tibial cartilage revealed an abundance of abnormal cells exhibiting numerous morphological changes. These resembled, but in some cases differed, from changes reported in classical apoptosis. The changes include abnormal distribution of chromatin, cell shrinkage, membrane blebbing and deposition of cell remnants (apoptotic bodies) in the lacuna space. Despite the TUNEL and EM changes, immunohistochemistry failed to detect any changes in the ratio of bax to bcl-2 in tibial chondrocytes of STR/ort mice. Both bcl-2 and bax levels decreased with age in morphologically normal STR/ort and control CBA cartilage. None of the caspases tested for was detected in tibial chondrocytes of either strain.


CONCLUSIONS
Chondrocyte cell death is correlated with the progression of OA in STR/ort mice and has many of the morphological characteristics of classical apoptosis. Absence of changes in bax to bcl-2 ratio in STR/ort chondrocytes indicate that the mitochondrial pathway of apoptosis is unlikely to be involved. Failure to detect caspases could be due to low levels of enzyme expression, expression within a very brief time period, or to a caspase-independent mechanism of cell death.
Mistry
D
D
Department of Experimental Biology, William Harvey Research Institute, St. Bartholomew's and The Royal London School of Medicine and Dentistry, Charterhouse Square, EC1M 6BQ, London, UK.
Oue
Y
Y
Chambers
M G
MG
Kayser
M V
MV
Mason
R M
RM
D000818:Animals; D017209:Apoptosis / Q000502:physiology*; D015415:Biomarkers / Q000032:analysis; D002358:Cartilage, Articular / Q000473:pathology / Q000503:physiopathology*; D053181:Caspase 8; D053453:Caspase 9; D020169:Caspases / Q000032:analysis; D019902:Chondrocytes / Q000502:physiology*; D004195:Disease Models, Animal; D006614:Hindlimb; D007150:Immunohistochemistry / Q000379:methods; D020287:In Situ Nick-End Labeling / Q000379:methods; D008297:Male; D051379:Mice; D008815:Mice, Inbred Strains; D008854:Microscopy, Electron / Q000379:methods; D010003:Osteoarthritis / Q000473:pathology / Q000503:physiopathology*; D011518:Proto-Oncogene Proteins / Q000032:analysis; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000032:analysis; D013977:Tibia; D051028:bcl-2-Associated X Protein
C494827:Bax protein, mouse; D015415:Biomarkers; D011518:Proto-Oncogene Proteins; D019253:Proto-Oncogene Proteins c-bcl-2; D051028:bcl-2-Associated X Protein; C505526:Casp8 protein, mouse; C506434:Casp9 protein, mouse; D053181:Caspase 8; D053453:Caspase 9; D020169:Caspases