Logo of biochemjBJ Latest papers and much more!
Biochem J. 2000 Sep 1; 350(Pt 2): 381–387.
PMCID: PMC1221264

Age-related accumulation of Maillard reaction products in human articular cartilage collagen.


Non-enzymic modification of tissue proteins by reducing sugars, the so-called Maillard reaction, is a prominent feature of aging. In articular cartilage, relatively high levels of the advanced glycation end product (AGE) pentosidine accumulate with age. Higher pentosidine levels have been associated with a stiffer collagen network in cartilage. However, even in cartilage, pentosidine levels themselves represent <1 cross-link per 20 collagen molecules, and as such cannot be expected to contribute substantially to the increase in collagen network stiffness. In the present study, we investigated a broad range of Maillard reaction products in cartilage collagen in order to determine whether pentosidine serves as an adequate marker for AGE levels. Not only did the well-characterized AGEs pentosidine, N(epsilon)-(carboxymethyl)lysine, and N(epsilon)-(carboxyethyl)lysine increase with age in cartilage collagen (all P<0.0001), but also general measures of AGE cross-linking, such as browning and fluorescence (both P<0.0001), increased. The levels of these AGEs are all higher in cartilage collagen than in skin collagen. As a functional measure of glycation the digestibility of articular collagen by bacterial collagenase was investigated; digestibility decreased linearly with age, proportional to the extent of glycation. Furthermore, the arginine content and the sum of the hydroxylysine and lysine content of cartilage collagen decrease significantly with age (P<0.0001 and P<0. 01 respectively), possibly due to modification by the Maillard reaction. The observed relationship between glycation and amino acid modification has not been reported previously in vivo. Our present results indicate that extensive accumulation of a variety of Maillard reaction products occurs in cartilage collagen with age. Altogether our results support the hypothesis that glycation contributes to stiffer and more brittle cartilage with advancing age.

Full Text

The Full Text of this article is available as a PDF (152K).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Dunn JA, Patrick JS, Thorpe SR, Baynes JW. Oxidation of glycated proteins: age-dependent accumulation of N epsilon-(carboxymethyl)lysine in lens proteins. Biochemistry. 1989 Nov 28;28(24):9464–9468. [PubMed]
  • Dunn JA, McCance DR, Thorpe SR, Lyons TJ, Baynes JW. Age-dependent accumulation of N epsilon-(carboxymethyl)lysine and N epsilon-(carboxymethyl)hydroxylysine in human skin collagen. Biochemistry. 1991 Feb 5;30(5):1205–1210. [PubMed]
  • Ahmed MU, Thorpe SR, Baynes JW. Identification of N epsilon-carboxymethyllysine as a degradation product of fructoselysine in glycated protein. J Biol Chem. 1986 Apr 15;261(11):4889–4894. [PubMed]
  • Wells-Knecht MC, Thorpe SR, Baynes JW. Pathways of formation of glycoxidation products during glycation of collagen. Biochemistry. 1995 Nov 21;34(46):15134–15141. [PubMed]
  • Sell DR, Monnier VM. Structure elucidation of a senescence cross-link from human extracellular matrix. Implication of pentoses in the aging process. J Biol Chem. 1989 Dec 25;264(36):21597–21602. [PubMed]
  • Uchiyama A, Ohishi T, Takahashi M, Kushida K, Inoue T, Fujie M, Horiuchi K. Fluorophores from aging human articular cartilage. J Biochem. 1991 Nov;110(5):714–718. [PubMed]
  • Bank RA, Bayliss MT, Lafeber FP, Maroudas A, Tekoppele JM. Ageing and zonal variation in post-translational modification of collagen in normal human articular cartilage. The age-related increase in non-enzymatic glycation affects biomechanical properties of cartilage. Biochem J. 1998 Feb 15;330(Pt 1):345–351. [PMC free article] [PubMed]
  • DeGroot J, Verzijl N, Bank RA, Lafeber FP, Bijlsma JW, TeKoppele JM. Age-related decrease in proteoglycan synthesis of human articular chondrocytes: the role of nonenzymatic glycation. Arthritis Rheum. 1999 May;42(5):1003–1009. [PubMed]
  • Ahmed MU, Brinkmann Frye E, Degenhardt TP, Thorpe SR, Baynes JW. N-epsilon-(carboxyethyl)lysine, a product of the chemical modification of proteins by methylglyoxal, increases with age in human lens proteins. Biochem J. 1997 Jun 1;324(Pt 2):565–570. [PMC free article] [PubMed]
  • Dyer DG, Blackledge JA, Thorpe SR, Baynes JW. Formation of pentosidine during nonenzymatic browning of proteins by glucose. Identification of glucose and other carbohydrates as possible precursors of pentosidine in vivo. J Biol Chem. 1991 Jun 25;266(18):11654–11660. [PubMed]
  • Schnider SL, Kohn RR. Effects of age and diabetes mellitus on the solubility and nonenzymatic glucosylation of human skin collagen. J Clin Invest. 1981 Jun;67(6):1630–1635. [PMC free article] [PubMed]
  • Monnier VM, Kohn RR, Cerami A. Accelerated age-related browning of human collagen in diabetes mellitus. Proc Natl Acad Sci U S A. 1984 Jan;81(2):583–587. [PMC free article] [PubMed]
  • Sell DR, Monnier VM. Isolation, purification and partial characterization of novel fluorophores from aging human insoluble collagen-rich tissue. Connect Tissue Res. 1989;19(1):77–92. [PubMed]
  • Lee KW, Simpson G, Ortwerth B. A systematic approach to evaluate the modification of lens proteins by glycation-induced crosslinking. Biochim Biophys Acta. 1999 Jan 6;1453(1):141–151. [PubMed]
  • Monnier VM, Sell DR, Nagaraj RH, Miyata S, Grandhee S, Odetti P, Ibrahim SA. Maillard reaction-mediated molecular damage to extracellular matrix and other tissue proteins in diabetes, aging, and uremia. Diabetes. 1992 Oct;41 (Suppl 2):36–41. [PubMed]
  • Dyer DG, Dunn JA, Thorpe SR, Bailie KE, Lyons TJ, McCance DR, Baynes JW. Accumulation of Maillard reaction products in skin collagen in diabetes and aging. J Clin Invest. 1993 Jun;91(6):2463–2469. [PMC free article] [PubMed]
  • Pokharna HK, Pottenger LA. Nonenzymatic glycation of cartilage proteoglycans: an in vivo and in vitro study. Glycoconj J. 1997 Dec;14(8):917–923. [PubMed]
  • Freeman MA. The fatigue of cartilage in the pathogenesis of osteoarthrosis. Acta Orthop Scand. 1975 Jun;46(3):323–328. [PubMed]
  • Weightman B. Tensile fatigue of human articular cartilage. J Biomech. 1976;9(4):193–200. [PubMed]
  • Kempson GE. Relationship between the tensile properties of articular cartilage from the human knee and age. Ann Rheum Dis. 1982 Oct;41(5):508–511. [PMC free article] [PubMed]
  • Degenhardt TP, Thorpe SR, Baynes JW. Chemical modification of proteins by methylglyoxal. Cell Mol Biol (Noisy-le-grand) 1998 Nov;44(7):1139–1145. [PubMed]
  • Schmidt MB, Mow VC, Chun LE, Eyre DR. Effects of proteoglycan extraction on the tensile behavior of articular cartilage. J Orthop Res. 1990 May;8(3):353–363. [PubMed]
  • Lyons TJ, Kennedy L. Non-enzymatic glycosylation of skin collagen in patients with type 1 (insulin-dependent) diabetes mellitus and limited joint mobility. Diabetologia. 1985 Jan;28(1):2–5. [PubMed]
  • Bank RA, Jansen EJ, Beekman B, te Koppele JM. Amino acid analysis by reverse-phase high-performance liquid chromatography: improved derivatization and detection conditions with 9-fluorenylmethyl chloroformate. Anal Biochem. 1996 Sep 5;240(2):167–176. [PubMed]
  • Bank RA, Beekman B, Verzijl N, de Roos JA, Sakkee AN, TeKoppele JM. Sensitive fluorimetric quantitation of pyridinium and pentosidine crosslinks in biological samples in a single high-performance liquid chromatographic run. J Chromatogr B Biomed Sci Appl. 1997 Dec 5;703(1-2):37–44. [PubMed]
  • Kim YJ, Sah RL, Doong JY, Grodzinsky AJ. Fluorometric assay of DNA in cartilage explants using Hoechst 33258. Anal Biochem. 1988 Oct;174(1):168–176. [PubMed]
  • Stegemann H, Stalder K. Determination of hydroxyproline. Clin Chim Acta. 1967 Nov;18(2):267–273. [PubMed]
  • Creemers LB, Jansen DC, van Veen-Reurings A, van den Bos T, Everts V. Microassay for the assessment of low levels of hydroxyproline. Biotechniques. 1997 Apr;22(4):656–658. [PubMed]
  • Takahashi M, Kushida K, Ohishi T, Kawana K, Hoshino H, Uchiyama A, Inoue T. Quantitative analysis of crosslinks pyridinoline and pentosidine in articular cartilage of patients with bone and joint disorders. Arthritis Rheum. 1994 May;37(5):724–728. [PubMed]
  • Maroudas A, Palla G, Gilav E. Racemization of aspartic acid in human articular cartilage. Connect Tissue Res. 1992;28(3):161–169. [PubMed]
  • Fu MX, Requena JR, Jenkins AJ, Lyons TJ, Baynes JW, Thorpe SR. The advanced glycation end product, Nepsilon-(carboxymethyl)lysine, is a product of both lipid peroxidation and glycoxidation reactions. J Biol Chem. 1996 Apr 26;271(17):9982–9986. [PubMed]
  • Miyata T, Inagi R, Asahi K, Yamada Y, Horie K, Sakai H, Uchida K, Kurokawa K. Generation of protein carbonyls by glycoxidation and lipoxidation reactions with autoxidation products of ascorbic acid and polyunsaturated fatty acids. FEBS Lett. 1998 Oct 16;437(1-2):24–28. [PubMed]
  • Eyre DR, Dickson IR, Van Ness K. Collagen cross-linking in human bone and articular cartilage. Age-related changes in the content of mature hydroxypyridinium residues. Biochem J. 1988 Jun 1;252(2):495–500. [PMC free article] [PubMed]
  • Miyahara T, Shiozawa S, Murai A. The effect of age on amino acid composition of human skin collagen. J Gerontol. 1978 Jul;33(4):498–503. [PubMed]
  • Dilley KJ, Harding JJ. Changes in proteins of the human lens in development and aging. Biochim Biophys Acta. 1975 Apr 29;386(2):391–408. [PubMed]
  • Coghlan SD, Augusteyn RC. Changes in the distribution of proteins in the aging human lens. Exp Eye Res. 1977 Dec;25(6):603–611. [PubMed]
  • Schnider SL, Kohn RR. Effects of age and diabetes mellitus on the solubility of collagen from human skin, tracheal cartilage and dura mater. Exp Gerontol. 1982;17(3):185–194. [PubMed]
  • Heaf DJ, Galton DJ. Sorbitol and other polyols in lens, adipose tissue and urine in diabetes mellitus. Clin Chim Acta. 1975 Aug 18;63(1):41–47. [PubMed]
  • Tomana M, Prchal JT, Garner LC, Skalka HW, Barker SA. Gas chromatographic analysis of lens monosaccharides. J Lab Clin Med. 1984 Jan;103(1):137–142. [PubMed]
  • Lee KW, Mossine V, Ortwerth BJ. The relative ability of glucose and ascorbate to glycate and crosslink lens proteins in vitro. off. Exp Eye Res. 1998 Jul;67(1):95–104. [PubMed]
  • Maroudas A. Distribution and diffusion of solutes in articular cartilage. Biophys J. 1970 May;10(5):365–379. [PMC free article] [PubMed]
  • Wells-Knecht MC, Huggins TG, Dyer DG, Thorpe SR, Baynes JW. Oxidized amino acids in lens protein with age. Measurement of o-tyrosine and dityrosine in the aging human lens. J Biol Chem. 1993 Jun 15;268(17):12348–12352. [PubMed]
  • Wells-Knecht MC, Lyons TJ, McCance DR, Thorpe SR, Baynes JW. Age-dependent increase in ortho-tyrosine and methionine sulfoxide in human skin collagen is not accelerated in diabetes. Evidence against a generalized increase in oxidative stress in diabetes. J Clin Invest. 1997 Aug 15;100(4):839–846. [PMC free article] [PubMed]
  • van Boekel MA. The role of glycation in aging and diabetes mellitus. Mol Biol Rep. 1991 May;15(2):57–64. [PubMed]
  • Brownlee M. Advanced protein glycosylation in diabetes and aging. Annu Rev Med. 1995;46:223–234. [PubMed]
  • Harding JJ, Dilley KJ. Structural proteins of the mammalian lens: a review with emphasis on changes in development, aging and cataract. Exp Eye Res. 1976 Jan;22(1):1–73. [PubMed]
  • Kern HL, Zolot SL. Transport of vitamin C in the lens. Curr Eye Res. 1987 Jul;6(7):885–896. [PubMed]
  • Taylor A, Jacques PF, Nadler D, Morrow F, Sulsky SI, Shepard D. Relationship in humans between ascorbic acid consumption and levels of total and reduced ascorbic acid in lens, aqueous humor, and plasma. Curr Eye Res. 1991 Aug;10(8):751–759. [PubMed]
  • Grushko G, Schneiderman R, Maroudas A. Some biochemical and biophysical parameters for the study of the pathogenesis of osteoarthritis: a comparison between the processes of ageing and degeneration in human hip cartilage. Connect Tissue Res. 1989;19(2-4):149–176. [PubMed]
  • Sims TJ, Rasmussen LM, Oxlund H, Bailey AJ. The role of glycation cross-links in diabetic vascular stiffening. Diabetologia. 1996 Aug;39(8):946–951. [PubMed]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society


Save items

Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...


  • Compound
    PubChem Compound links
  • MedGen
    Related information in MedGen
  • PubMed
    PubMed citations for these articles
  • Substance
    PubChem Substance links

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...