Crosslinking by advanced glycation end products increases the stiffness of the collagen network in human articular cartilage: a possible mechanism through which age is a risk factor for osteoarthritis.
OBJECTIVE
Age is an important risk factor for osteoarthritis (OA). During aging, nonenzymatic glycation results in the accumulation of advanced glycation end products (AGEs) in cartilage collagen. We studied the effect of AGE crosslinking on the stiffness of the collagen network in human articular cartilage.


METHODS
To increase AGE levels, human adult articular cartilage was incubated with threose. The stiffness of the collagen network was measured as the instantaneous deformation (ID) of the cartilage and as the change in tensile stress in the collagen network as a function of hydration (osmotic stress technique). AGE levels in the collagen network were determined as: Nepsilon-(carboxy[m]ethyl)lysine, pentosidine, amino acid modification (loss of arginine and [hydroxy-]lysine), AGE fluorescence (360/460 nm), and digestibility by bacterial collagenase.


RESULTS
Incubation of cartilage with threose resulted in a dose-dependent increase in AGEs and a concomitant decrease in ID (r = -0.81, P < 0.001; up to a 40% decrease at 200 mM threose), i.e., increased stiffness, which was confirmed by results from the osmotic stress technique. The decreased ID strongly correlated with AGE levels (e.g., AGE fluorescence r = -0.81, P < 0.0001). Coincubation with arginine or lysine (glycation inhibitors) attenuated the threose-induced decrease in ID (P < 0.05).


CONCLUSIONS
Increasing cartilage AGE crosslinking by in vitro incubation with threose resulted in increased stiffness of the collagen network. Increased stiffness by AGE crosslinking may contribute to the age-related failure of the collagen network in human articular cartilage to resist damage. Thus, the age-related accumulation of AGE crosslinks presents a putative molecular mechanism whereby age is a predisposing factor for the development of OA.
Verzijl
Nicole
N
TNO Prevention and Health, Leiden, The Netherlands.
DeGroot
Jeroen
J
Ben
Zaken Chaya
ZC
Brau-Benjamin
Orit
O
Maroudas
Alice
A
Bank
Ruud A
RA
Mizrahi
Joe
J
Schalkwijk
Casper G
CG
Thorpe
Suzanne R
SR
Baynes
John W
JW
Bijlsma
Johannes W J
JW
Lafeber
Floris P J G
FP
TeKoppele
Johan M
JM
D000328:Adult; D000375:Aging / Q000378:metabolism / Q000473:pathology; D001120:Arginine / Q000378:metabolism / Q000494:pharmacology; D002358:Cartilage, Articular / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology*; D003094:Collagen / Q000378:metabolism*; D003432:Cross-Linking Reagents / Q000378:metabolism*; D017127:Glycation End Products, Advanced / Q000378:metabolism*; D006801:Humans; D008239:Lysine / Q000378:metabolism / Q000494:pharmacology; D010003:Osteoarthritis / Q000378:metabolism / Q000473:pathology; D011765:Pyruvaldehyde / Q000494:pharmacology; D012044:Regression Analysis; D012266:Ribose / Q000494:pharmacology; D013314:Stress, Mechanical; D013780:Tetroses / Q000494:pharmacology
D003432:Cross-Linking Reagents; D017127:Glycation End Products, Advanced; D013780:Tetroses; D012266:Ribose; D011765:Pyruvaldehyde; D003094:Collagen; D001120:Arginine; D008239:Lysine; C073321:erythrose