Destruction of articular cartilage is a characteristic feature of osteoarthritis (OA). Collagen hydrolysates are mixtures of collagen peptides and have gained huge public attention as nutriceuticals used for prophylaxis of OA. Here, we evaluated for the first time whether different bovine collagen hydrolysate preparations indeed modulate the metabolism of collagen and proteoglycans from human OA cartilage explants and determined the chemical composition of oligopeptides representing collagen fragments. Using biophysical techniques, like MALDI-TOF-MS, AFM, and NMR, the molecular weight distribution and aggregation behavior of collagen hydrolysates from bovine origin (CH-Alpha?, Peptan? B 5000, Peptan? B 2000) were determined. To investigate the metabolism of human femoral OA cartilage, explants were obtained during knee replacement surgery. Collagen synthesis of explants as modulated by 0–10 mg/ml collagen hydrolysates was determined using a novel dual radiolabeling procedure. Proteoglycans, NO, PGE2, MMP-1, -3, -13, TIMP-1, collagen type II, and cell viability were determined in explant cultures. Groups of data were analyzed using ANOVA and the Friedman test (n = 5–12). The significance was set to p≤0.05. We found that collagen hydrolysates obtained from different sources varied with respect to the width of molecular weight distribution, average molecular weight, and aggregation behavior. None of the collagen hydrolysates tested stimulated the biosynthesis of collagen. Peptan? B 5000 elevated NO and PGE2 levels significantly but had no effect on collagen or proteoglycan loss. All collagen hydrolysates tested proved not to be cytotoxic. Together, our data demonstrate for the first time that various collagen hydrolysates differ with respect to their chemical composition of collagen fragments as well as by their pharmacological efficacy on human chondrocytes. Our study underscores the importance that each collagen hydrolysate preparation should first demonstrate its pharmacological potential both in vitro and in vivo before being used for both regenerative medicine and prophylaxis of OA.
References
[1]
Oesser S, Adam M, Babel W, Seifert J (1999) Oral administration of 14C-labeled gelatin hydrolysate leads to an accumulation of radioactivity in cartilage of mice (C57/BL). J Nutr 129: 1891–1895.
[2]
European Food Safety Authority (2005) Opinion of the Food Safety Authority on safety of collagen and a processing method for the production of collagen. EFSA J 174: 1–9.
[3]
Bello AE, Oesser S (2006) Collagen hydrolysate for the treatment of osteoarthritis and other joint disorders: a review of the literature. Curr Med Res Opin 22: 2221–2232.
[4]
Clark KL, Sebastianelli W, Flechsenhar KR, Aukermann DF, Meza F, et al. (2008) 24-week study on the use of collagen hydrolysate as a dietary supplement in athletes with activity-related joint pain. Curr Med Res Opin 24: 1485–1496.
[5]
Moskowitz RW (2000) Role of collagen hydrolysate in bone and joint disease. Semin Arthritis Rheum 30: 87–99.
[6]
Benito-Ruiz P, Camacho-Zambrano MM, Carrillo-Arcentales JN, Mestanza-Peralta Ma, Vallejo-Flores CA, et al. (2009) A randomized controlled trial on the efficacy and safety of a food ingredient, collagen hydrolysate, for improving joint comfort. Int J Food Sci Nutr (Suppl 2): 99–113.
[7]
Henroitin Y, Lambert C, Couchourel D, Ripoll C, Chiotelli E (2011) Nutraceuticals: do they represent a new era in the management of osteoarthritis? – A narrative review from the lessons taken with five products. Osteoarthritis Cartilage 19: 1–21.
[8]
McAlindon TE, Muite M, Krishnan N, Ruthazer R, Price LL, et al. (2011) Change in knee osteoarthritis cartilage detected by delayed gadolinium enhanced magnetic resonance imaging following treatment with collagen hydrolysate: a pilot randomized controlled trial. Osteoarthritis Cartilage 19: 399–405.
[9]
EFSA Panel on Dietetic Products, Nutrition and Allergies (2011) Scientific Opinion on the substantiation of a health claim related to collagen hydrolysate and maintenance of joints pursuant to Article 13(5) of Regulation (EC) No 1924/2006. EFSA Journal 9(7): 2291.
[10]
Oesser S, Seifert J (2003) Stimulation of type II collagen biosynthesis and secretion in bovine chondrocytes cultured with degraded collagen. Cell Tissue Res 311: 393–399.
[11]
Raabe O, Reich C, Wenisch S, Hild A, Burg-Roderfeld M, et al. (2010) Hydrolyzed fish collagen induced chondrogenic differentiation of equine adipose tissue-derived stromal cells. Histochem Cell Biol 134: 545–554.
[12]
Jennings L, Wu L, King KB, H?mmerle H, Cs-Szabo G, et al. (2001) The effect of collagen fragments on the extracellular matrix metabolism of bovine and human chondrocytes. Connect Tissue Res 42: 71–86.
[13]
Fichter M, K?rner U, Sch?mburg J, Jennings L, Cole AA, et al. (2006) Collagen degradation products modulate matrix metalloproteinase expression in cultured articular chondrocytes. J Orthop Res 24: 63–70.
[14]
Siebert H-C, Burg-Roderfeld M, Eckert T, St?tzel S, Kirch U, et al. (2010) Interaction of the α2A domain of integrin with small collagen fragments. Protein & Cell 1(4): 393–405.
[15]
St?tzel S, Schurink M, Wienk H, Siebler U, Burg-Roderfeld M, et al. (2012) Molecular organization of different collagen hydrolysates and collagen fragments as revealed by a combination of Atomic Force Microscopy (AFM) and Diffusion Ordered NMR Spectroscopy (DOSY). ChemPhysChem 13: 3117–3125.
[16]
Goldring MB (2006) Update on the biology of the chondrocytes and new approaches to treating cartilage diseases. Best Pract Res Clin Rheumatol 20: 1003–1025.
[17]
Roman-Blas JA, Jimenez SA (2006) NF-kB as a potential therapeutic target in osteoarthritis and rheumatoid arthritis. Osteoarthritis Cartilage 14: 839–848.
[18]
Goldring MB, Berenbaum F (2004) The regulation of chondrocyte function by proinflammatory mediators: prostaglandins and nitric oxide. Clin Orthop Relat Res 427 (Suppl): S37–46.
[19]
Attur M, Al-Mussawir HE, Patel J, Kitay A, Dave M, et al. (2008) Prostaglandin E2 exerts catabolic effects in osteoarthritis cartilage: evidence for signaling via the EP4 receptor. J Immunol 181: 5082–5088.
[20]
Fukuda K, Kumano F, Takayama M, Saito M, Otani K, et al. (1995) Zonal differences in nitric oxide synthesis by bovine chondrocytes exposed to interleukin-1. Inflamm Res 44: 434–437.
[21]
Steinmeyer J, Konttinen YT (2006) Oral treatment options for degenerative joint disease-presence and future. Adv Drug Del Rev 58: 168–211.
[22]
Bayliss MT, Ali SY (1979) Age-related changes in the composition and structure of human articular cartilage proteoglycans. Biochem J 176: 683–693.
[23]
Fan Z, Bau B, Yang H, Soeder S, Aigner T (2005) Freshly isolated osteoarthritic chondrocytes are catabolically more active than normal chondrocytes, but less responsive to catabolic stimulation with interleukin-1beta. Arthritis Rheum 52(1): 136–143.
[24]
Aigner T, Haag J, Martin J, Buckwalter J (2007) Osteoarthritis: aging of matrix and cells–going for a remedy. Curr Drug Targets 8(2): 325–332.
[25]
Collins DH (1949) The Pathology of articular and spinal diseases. London: Edward Arnold and Co, 76–79.
[26]
Sadowski T, Steinmeyer J (2001) Effects of tetracyclines on the production of matrix metalloproteinases and plasminogen activators as well as of their natural inhibitors, tissue inhibitor of metalloproteinases-1 and plasminogen activator inhibitor-1. Inflamm Res 50: 175–182.
[27]
Sadowski T, Steinmeyer J (2001) Minocycline inhibits the production of inducible nitric oxide synthase in articular chondrocytes. J Rheumatol 28: 336–340.
[28]
Steinmeyer J, Kordelle J, Stürz H (2010) In vitro inhibition of aggrecanase activity by tetracyclines and proteoglycan loss from osteoarthritic human articular cartilage. J Orthop Res 28: 828–833.
[29]
Goodwin JL, Farley ML, Swaim B, Goldring SR, Goldring MB, et al. (2008) Dual proline labeling protocol for individual “baseline” and “response” biosynthesis measurements in human articular cartilage. Osteoarthritis Cartilage 16: 1263–1266.
[30]
Juva K, Prockop DJ (1966) Modified procedure for the assay of H3– or C14-labeled hydroxyproline. Anal Biochem 15: 77–83.
[31]
Switzer BR, Summer GK (1971) Improved method for hydroxyproline analysis in tissue hydrolysates. Anal Biochem 39: 487–491.
[32]
Farndale RW, Buttle DJ, Barrett AJ (1986) Improved quantitation and discrimination of sulphated glycosaminoglycans by use of dimethylmethylene blue. Biochim Biophys Acta 83: 173–177.
[33]
Green LC, Wagner DA, Glogowski J, Skipper PL, Wishnok JS (1982) Analysis of nitrate, nitrite, and [15N]nitrate in biological fluids. Anal Biochem 126: 131–138.
[34]
Sauerland K, Raiss RX, Steinmeyer J (2003) Proteoglycan metabolism and viability of articular cartilage explants as modulated by the frequency of intermittent loading. Osteoarthritis Cartilage 11: 343–350.
[35]
Aigner T, Zien A, Ersatz A, Gebhard PM, McKenna L (2001) Anabolic and catabolic gene expression pattern analysis in normal versus osteoarthritic cartilage using complementary DNA-array technology. Arthritis Rheum 44: 2777–2789.
[36]
Aigner T, Fundel K, Saas J, Gebhard PM, Haag J, et al. (2006) Large-scale gene expression profiling reveals major pathogenetic pathways of cartilage degeneration in osteoarthritis. Arthritis Rheum 54: 3533–3544.
[37]
Eckert T, St?tzel S, Burg-Roderfeld M, Sewing J, Lütteke T, et al. (2012) In silico study on sulfated and non-sulfated carbohydrate chains from proteoglycans in Cnidaria and their interaction with collagen. O J Phys Chem, in press.
[38]
Oesser S, Proksch E, Schunck M (2008) Prophylactic treatment with a special collagen hydrolysate decreases cartilage tissue degeneration in the knee joints. Osteoarthritis Cartilage (Suppl 4): S45.
[39]
Wolf A, Ackermann B, Steinmeyer J (2007) Collagen synthesis of articular cartilage explants in response to frequency of cyclic mechanical loading. Cell Tissue Res 327: 155–166.
[40]
Hollander AP, Pidoux I, Rainer A, Rorabeck C, Bourne R, et al. (1995) Damage of type II collagen in aging and osteoarthritis starts at the articular surface, originates around chondrocytes, and extends into the cartilage with progressive degeneration. J Clin Invest 96: 2859–2869.
[41]
Bank RA, Krikken M, Beekman B, Stoop R, Maroudas A, et al. (1997) A simplified measurement of degraded collagen in tissues: application in healthy, fibrillated and osteoarthritic cartilage. Matrix Biol 16: 233–243.
[42]
Naito S, Shiomi T, Okada A, Kimura T, Chijiwa M, et al. (2007) Expression of ADAMTS4 (aggreacanse-1) in human osteoarthritic cartilage. Pathol Int 57: 703–711.
[43]
Hashimoto G, Aoki T, Nakamura H, Tanzawa K, Okada Y (2001) Inhibition of ADAMTS4 (aggrecanase-1) by tissue inhibitors of metalloproteinases (TIMP-1, 2, 3, 4). FEBS Lett 494: 192–195.
[44]
Kashiwagi M, Tortorella M, Nagase H, Brew K (2001) TIMP-3 is a potent inhibitor of aggrecanase 1 (ADAM-TS4) and aggrecanase 2 (ADAM-TS5). J Biol Chem 276: 12501–12504.
[45]
Vincent TL, Saklatvala J (2008) Is the response of cartilage to injury relevant to osteoarthritis? Arthritis Rheum 58: 1207–1210.
[46]
Fukui N, Ikeda Y, Ohnuki T, Tanaka N, Hikita A, et al. (2008) Regional differences in chondrocyte metabolism in osteoarthritis: a detailed analysis by laser capture microdissection. Arthritis Rheum 58: 154–163.