title: 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.
abstract: 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.
authors.affiliation: TNO Prevention and Health, Leiden, The Netherlands.
authors.forename: Nicole
authors.initials: N
authors.lastname: Verzijl
authors.forename: Jeroen
authors.initials: J
authors.lastname: DeGroot
authors.forename: Zaken Chaya
authors.initials: ZC
authors.lastname: Ben
authors.forename: Orit
authors.initials: O
authors.lastname: Brau-Benjamin
authors.forename: Alice
authors.initials: A
authors.lastname: Maroudas
authors.forename: Ruud A
authors.initials: RA
authors.lastname: Bank
authors.forename: Joe
authors.initials: J
authors.lastname: Mizrahi
authors.forename: Casper G
authors.initials: CG
authors.lastname: Schalkwijk
authors.forename: Suzanne R
authors.initials: SR
authors.lastname: Thorpe
authors.forename: John W
authors.initials: JW
authors.lastname: Baynes
authors.forename: Johannes W J
authors.initials: JW
authors.lastname: Bijlsma
authors.forename: Floris P J G
authors.initials: FP
authors.lastname: Lafeber
authors.forename: Johan M
authors.initials: JM
authors.lastname: TeKoppele
mesh_terms: 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
chemical_list: D003432:Cross-Linking Reagents; D017127:Glycation End Products, Advanced; D013780:Tetroses; D012266:Ribose; D011765:Pyruvaldehyde; D003094:Collagen; D001120:Arginine; D008239:Lysine; C073321:erythrose