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 naïve 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 naïve knees showed no difference as compared with the basal values. The concentrations (microM) of aspartate and glutamate in knee dialysates at 30 weeks after ACLT in naïve, 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 naïve 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).


CONCLUSIONS
Our findings indicate an involvement of EAAs and EAATs in the pathogenesis of OA in ACLT rabbits.
Section of Orthopedic Surgery, Pingtung Christian Hospital, Pingtung, Taiwan.
Y-H
YH
Jean
Z-H
ZH
Wen
Y-C
YC
Chang
S-P
SP
Hsieh
J-D
JD
Lin
C-C
CC
Tang
W-F
WF
Chen
A-K
AK
Chou
C-S
CS
Wong
D000818:Animals; D016118:Anterior Cruciate Ligament / Q000737:chemistry*; D001224:Aspartic Acid / Q000378:metabolism*; D002985:Clinical Protocols; D018846:Excitatory Amino Acids / Q000378:metabolism*; D018698:Glutamic Acid / Q000378:metabolism*; D007150:Immunohistochemistry; D007270:Injections, Intra-Articular; D007719:Knee Joint / Q000601:surgery; D008297:Male; D026901:Membrane Transport Proteins / Q000737:chemistry; D017551:Microdialysis; D020370:Osteoarthritis, Knee / Q000378:metabolism* / Q000503:physiopathology; D011817:Rabbits; D016059:Range of Motion, Articular / Q000502:physiology
D018846:Excitatory Amino Acids; D026901:Membrane Transport Proteins; D001224:Aspartic Acid; D018698:Glutamic Acid