Increase in excitatory amino acid concentration and transporters expression in osteoarthritic knees of anterior cruciate ligament transected rabbits

Objective: The present study aimed to determine the role of excitatory amino acids (EAAs) and EAA transporters (EAATs) in an osteoarthritis (OA) model of rabbit knees. Methods: OA was induced in New Zealand white male rabbits by anterior cruciate ligament transection (ACLT) in one knee of one hind limb; the other knee left unoperated. Rabbits that received ACLT of knee were assigned to the ACLT group (n = 6), while a sham-operated group (n = 6) underwent arthrotomy with no ACLT. Six naive rabbits that received no surgery were used as normal control. The width of the knee joint was measured to determine the severity of joint inflammation. Before operation and at 10, 20, and 30 weeks after operation, knee joint dialysates were collected by microdialysis and assayed for EAAs by high-performance liquid chromatography. Gross morphology and histopathology and EAATs glutamate/aspartate transporter (GLAST) and glutamate transporter-1 (GLT-1) expression in the articular cartilage of the knees were evaluated by immunohistochemistry and western blot analysis. Results: In the ACLT knees, a significant increase in the joint width was observed (5.3 +/- 0.9 mm, P < 0.05) at 30 weeks after operation, while the sham-operated and naive knees showed no difference as compared with the basal values. The concentrations (mu M) of aspartate and glutamate in knee dialysates at 30 weeks after ACLT in naive, sham, and ACLT were 0.36 +/- 0.07 and 4.5 +/- 1.10; 0.38 +/- 0.09 and 4.61 +/- 1.11; 0.67 +/- 0.18 and 9.71 +/- 2.89, respectively. Levels of glutamate and aspartate in the dialysates obtained from the ACLT knees increased by 213.3 +/- 29.6% and 187.5 +/- 33.8% (P < 0.05) when compared to those in the sham-operated knees. Both naive and ACLT chondrocytes were positively stained by antibodies against GLAST and GLT-1. GLAST and GLT-1 protein expressions were significantly increased in the ACLT knees (P < 0.05). Conclusion: Our findings indicate an involvement of EAAs and EAATs in the pathogenesis of OA in ACLT rabbits. (C) 2008 Osteoarthritis Research Society International. Published by Elsevier Ltd. All rights reserved.