The inhibition of collagenase induced degradation of collagen by the galloyl-containing polyphenols tannic acid, epigallocatechin gallate and epicatechin gallate

  • John K. JacksonEmail author
  • Jinying Zhao
  • Wesley Wong
  • Helen M. Burt


Collagen based cosmetic fillers require repeat treatments due to collagenase derived degradation of the filler in the intradermal injection site. The objective of this study was to investigate the inhibition of this degradation by the galloyl-containing compounds tannic acid, epigallocatechin gallate (EGCG), epicatechin gallate (ECG) and gallic acid (GA). A gel permeation chromatography assay was developed to quantitate the collagenase induced reductions in collagen molecular weight. The binding of the compounds to collagen was measured using HPLC. The stabilization of collagen was measured using Differential Scanning Calorimetry (DSC). Tannic acid, EGCG and ECG (but not GA) were found to strongly inhibit collagen degradation at concentrations in the low micromolar range. The compounds bound strongly to collagen and stabilized collagen. It is concluded that tannic acid, EGCG and ECG bind to collagen via extensive hydrogen bonding augmented by some hydrophobic interactions and prevent the free access of collagenase to active sites on the collagen chains.


Hyaluronic Acid Polyphenol Collagenase Gallic Acid EGCG 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.



This work was supported using funds from Angiotech Pharmaceuticals, Station Street, Vancouver, BC, Canada.


  1. 1.
    Goldberg DJ. Fillers in cosmedic dermatology. London, PA: Informa Healthcare; 2006.Google Scholar
  2. 2.
    Eppley BL, Dadvand B. Injectable soft-tissue fillers: clinical overview. Plast Reconstr Surg. 2006;118(4):98e–106e.CrossRefPubMedGoogle Scholar
  3. 3.
    Werschler WP, Weinkle S. Longevity of effects of injectable products for soft-tissue augmentation. J Drugs Dermatol. 2005;4(1):20–7.PubMedGoogle Scholar
  4. 4.
    Baumann L, Kaufman J, Saghari S. Collagen fillers. Dermatol Ther. 2006;19:134–40.CrossRefPubMedGoogle Scholar
  5. 5.
    Owens JM. Soft tissue implants and fillers. Otolaryngol Clin North Am. 2005;38:361–9.CrossRefPubMedGoogle Scholar
  6. 6.
    Laeschke K. Biocompatibility of microparticles into soft tissue fillers. Semin Cutan Med Surg. 2004;23:214–7.CrossRefPubMedGoogle Scholar
  7. 7.
    Pitaru S, Noff M, Blok L, et al. Long-term efficacy of novel ribose-cross-linked collagen dermal filler: a histologic and histomorphometric study in an animal model. Dermatol Surg. 2007;33:1045–54.CrossRefPubMedGoogle Scholar
  8. 8.
    Covington AD, Lilley TH, Song L, Evans CS. Collagen and polyphenols: new relationships and new outcomes. Part 1. Flavonoid reactions for new tanning processes. Jalca. 2005;100:325–43.Google Scholar
  9. 9.
    Deaville ER, Green RJ, Mueller-Harvey I, Willoughby I, Frazier RA. Hydrolyzable tannin structures influence relative globular and random coil protein binding strengths. J Agric Food Chem. 2007;55:4454–561.CrossRefGoogle Scholar
  10. 10.
    Hagerman AE, Butler L. Protein precipitation method for the quantitative determination of tannins. J Agric Food Chem. 1978;26(4):809–11.CrossRefGoogle Scholar
  11. 11.
    Goo HC, Hwang YS, Choi YR, Cho HN, Suh H. Development of collagenase-resistant collagen and its interaction with adult human dermal fibroblasts. Biomaterials. 2003;24:5099–113.CrossRefPubMedGoogle Scholar
  12. 12.
    Liao XP, Lu ZB, Shi B. Selective adsorption of vegetable tannins onto collagen fibers. Ind Eng Chem Res. 2003;42:3397–402.CrossRefGoogle Scholar
  13. 13.
    Tang HR, Covington AD, Hancock RA. Structure-activity relationships in the hydrophobic interactions of polyphenols with cellulose and collagen. Biopolymers. 2003;70(3):403–13.CrossRefPubMedGoogle Scholar
  14. 14.
    Demule M, Brossard M, Pagé M, Gingras D, Béliceau R. Matrix metalloproteinase inhibition by green tea catechins. Biochim Biophys Acta. 2000;1478:51–60.Google Scholar
  15. 15.
    Deyl Z, Macek K. High-performance gel permeation chromatography of collagens. J Chromatogr. 1982;230:409–14.CrossRefPubMedGoogle Scholar
  16. 16.
    Meyer M, Morgenstern B. Characterization of Gelatine and acid soluble collagen by size exclusion chromatography coupled with multi angle light scattering (SEC-MALS). Biomacromolecules. 2003;4:1727–32.CrossRefPubMedGoogle Scholar
  17. 17.
    Saito ST, Froelich PE, Gosaman G, Bergold AM. Full validation of a simple method for determination of catechins and caffeine in Brazilian green tea (Cameillia sinensis assamica) using HPLC. Chrmoatographia. 2007;65:607–10.Google Scholar
  18. 18.
    Xu J, Tan T, Janson JC. One step purification of epigallocatechin gallate from crude green tea extracts by mixed mode adsorption chromatography on highly crosslinked agarose media. J Chromatogr A. 2007;1169:235–8.CrossRefPubMedGoogle Scholar
  19. 19.
    Hong Jungil, Lu Hong, Meng Xiaofeng, Ryu Jae-Ha, Hara Yukihiko, Yang ChungS. Stability, Cellular uptake, biotransformation, and efflux of tea polyphenol (-)-Epigallocatechin-3-Gallate in HT-29 human colon adenocarcinoma cells. Cancer Res. 2002;62:7241–6.PubMedGoogle Scholar
  20. 20.
    Peterson JT. Matrix metalloproteinase inhibitor development and the remodelling of drug discovery. Heart Fail Rev. 2004;9:63–79.CrossRefPubMedGoogle Scholar
  21. 21.
    Covington AD. Modern tanning chemistry. Chem Soc Rev. 1997;26:111–26.CrossRefGoogle Scholar
  22. 22.
    Calderon P, Van Buren J, Robinson WB. Factors influencing the formation of precipitates and hazes by gelatine and condensed and hydrolysable tannins. J Agric Food Chem. 1968;16(3):479–82.CrossRefGoogle Scholar
  23. 23.
    Cho HH, Matsumura K, Nakajima N, Han DW, Tsutsumi S, Hyon SH. Degradation control of collagen by epigallocatechin-3-o-gallate. Key Eng Mater. 2007;342–343:781–4.CrossRefGoogle Scholar
  24. 24.
    Makimura M, Hirasawa M, Kobayashi K, et al. Inhibitory effect of tea catechins on collagenase activity. J Periodontol. 1993;7:630–6.Google Scholar
  25. 25.
    Celej MS, Montich GG, Fidelio GD. Protein stability induced by ligand binding correlates with changes in protein flexibility. Protein Sci. 2003;12:1496–506.CrossRefPubMedGoogle Scholar
  26. 26.
    Fathima NN, Madhan B, Rao JR, Nair BU, Ramasami T. Interaction of aldehydes with collagen: effect of thermal, enzymatic and conformational stability. Int J Biol Macromol. 2004;34:241–7.CrossRefPubMedGoogle Scholar
  27. 27.
    Tang HR, Covington AD, Hancock RA. Use of DSC to detect the heterogeneity of hydrothermal stability in the polyphenol-treated collagen matrix. J Agric Food Chem. 2003;51:6652–6.CrossRefPubMedGoogle Scholar
  28. 28.
    McClain P, Wiley E. Differential scanning calorimeter studies of the thermal transitions of collagen. J Biol Chem. 1972;247(8):692–7.PubMedGoogle Scholar
  29. 29.
    Miles CA, Avery NC, Rodin VV, Bailey AJ. The increase in denaturation temperature following cross-linking of collagen is caused by dehydration of the fibers. J Mol Biol. 2005;346:551–6.CrossRefPubMedGoogle Scholar
  30. 30.
    Cotran RS, Kumar V, Robbins SL. Pathologic basis of disease. Philadelphia: W.B. Saunders Company; 1989. p. 39–87.Google Scholar
  31. 31.
    Seandel M, Noack-Kunnmann K, Zhu D, Aimes RT, Quigley JP. Growth factor induced angiogenesis in vivo requires specific cleavage of fibrillar type 1 collagen. Blood. 2001;97:2323–32.CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  • John K. Jackson
    • 1
    Email author
  • Jinying Zhao
    • 1
  • Wesley Wong
    • 1
  • Helen M. Burt
    • 1
  1. 1.Faculty of Pharmaceutical SciencesUniversity of British ColumbiaVancouverCanada

Personalised recommendations