The utility of measuring C-terminal telopeptides of collagen type II (CTX-II) in serum and synovial fluid samples for estimation of articular cartilage status in experimental models of destructive joint diseases.
OBJECTIVE
To characterize and validate a novel, enzyme-linked immunoassay for measuring cross-linked dimer forms of C-terminal telopeptides of type II collagen (CTX-II) in serum and synovial fluid of rodents, and investigate whether CTX-II measurements can reflect joint status in two established animal models of destructive joint diseases.


METHODS
Firstly, the specificity, in vivo validity, antigen recovery, and reproducibility of the assay were investigated. Secondly, we induced arthritis in rats using either bovine collagen type II or mono-iodoacetate. CTX-II levels were measured in the serum and synovial fluid of the affected femoro-tibial joint and correlated with microscopic severity of joint lesions as determined by validated scoring systems.


RESULTS
The F4601 monoclonal antibody (mAb) is highly specific for the EKGPDP sequence at the CTX-II. Strong CTX-II signals were detected during enzymatic degradation of articular cartilage explants by matrix metalloproteinase (MMP)-9 or MMP-13. The assay presented a good degree of precision and reproducibility (inter- and intra-assay CVs< 8.0%). In the collagen-induced arthritis (CIA) model, the assay indicated markedly increased levels of CTX-II in both the synovial fluid and the serum. Furthermore, CTX-II levels in both the synovial fluid (r = 0.76; P < 0.0001) and the serum (r = 0.85; P < 0.0001) showed strong correlations with the microscopic severity scores of joint lesions at Day 22. In the mono-iodoacetate-induced arthritis (MIA) model, CTX-II concentration in the synovial fluid (r = 0.53; P < 0.0001), but not in the serum, correlated with the microscopic severity score.


CONCLUSIONS
The Preclinical CTX-II assay could provide a useful supplement to currently available methods for the non-invasive assessment of cartilage status. The utility of serum CTX-II to reflect joint status appeared to be limited to systemic forms of destructive joint diseases.
Oestergaard
S
S
Nordic Bioscience A/S, Herlev, Denmark. soe@nordicbioscience.com
Chouinard
L
L
Doyle
N
N
Karsdal
M A
MA
Smith
S Y
SY
Qvist
P
P
Tankó
L B
LB
D000818:Animals; D001169:Arthritis, Experimental / Q000097:blood / Q000473:pathology; D002358:Cartilage, Articular / Q000473:pathology*; D024042:Collagen Type I / Q000097:blood / Q000737:chemistry; D024043:Collagen Type II / Q000097:blood / Q000737:chemistry; D004195:Disease Models, Animal; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods / Q000592:standards; D007596:Joints / Q000473:pathology; D010455:Peptides / Q000097:blood / Q000737:chemistry; D051381:Rats; D015203:Reproducibility of Results; D013582:Synovial Fluid / Q000737:chemistry
D024042:Collagen Type I; D024043:Collagen Type II; D010455:Peptides; C052929:collagen type I trimeric cross-linked peptide