Cellular events leading to chondrocyte death after cartilage impact injury.
OBJECTIVE
We undertook this study to test our postulate that leukocytes extend the zone of injury in cartilage after acute mechanical trauma.


METHODS
Fresh cadaveric canine femoral condyles were subjected to 20-25-MPa impact injury. Condyle explants or dispersed chondrocytes were cultured with autologous blood mononuclear leukocytes (MNLs). Viability of chondrocytes at varying distances from the impact site was assessed by trypan blue exclusion.


RESULTS
Mechanical injury caused a significant loss of viable chondrocytes over 7 days, even in cartilage >10 mm from the impact site. After biomechanical stress, death of cells within 10 mm of the impact could be largely prevented by addition of N(G)-monomethyl-L-arginine to inhibit nitric oxide (NO) generation. Chondrocytes within 10 mm of the impact were also susceptible to killing by living MNLs, but not by incubation with the supernatants of endotoxin-activated MNLs. Chondrocytes in this vulnerable zone expressed intercellular adhesion molecule 1 (ICAM-1) (CD54), facilitating attachment of MNLs that localized adjacent to the chondrocytes. Leukocytes killed dispersed chondrocytes harvested from the impact zone by generation of reactive oxygen species. Leukocyte-mediated killing could be blocked by desferoxamine or by antibodies to CD18, which prevent attachment of leukocytes to ICAM-1-expressing chondrocytes.


CONCLUSIONS
Our data suggest that after mechanical injury, chondrocytes distant from the site may be killed through the generation of NO. Inflammatory leukocytes further extend the zone of chondrocyte death by adhering to chondrocytes expressing ICAM-1 and by inducing the accumulation of free oxygen radicals in the chondrocyte cytoplasm. Patients may benefit from therapies that reduce infiltration of inflammatory leukocytes into acutely injured cartilage.
Green
D M
DM
Baylor College of Medicine, Houston, Texas, USA.
Noble
P C
PC
Ahuero
J S
JS
Birdsall
H H
HH
D000818:Animals; D002356:Cartilage / Q000293:injuries* / Q000473:pathology*; D016923:Cell Death; D019902:Chondrocytes / Q000473:pathology*; D004285:Dogs; D007963:Leukocytes, Mononuclear / Q000502:physiology