Knee Surgery, Sports Traumatology, Arthroscopy

, Volume 20, Issue 6, pp 1159–1167 | Cite as

Expression levels of matrix metalloproteinase (MMP)-9 and its specific inhibitor TIMP-1, in septic and aseptic arthritis of the knee

  • Vasileios Christos Fotopoulos
  • Athina Tzinia
  • Matthew Tzurbakis
  • Vasiliki Kalfakakou
  • Stamatina Levidiotou-Stefanou
  • Anastasios Georgoulis



In cases of septic knee arthritis, there is excess of matrix metalloproteinases (MMPs) over tissue inhibitors of metalloproteinases (TIMPs), due to enhanced expression and activation that are induced by bacteria in comparison with rheumatic or degenerative arthritis. The aim of this study was to explore the expression levels of synovial gelatinase MMP-9 and its specific inhibitor TIMP-1 in septic and aseptic arthritis and their potential use as additional aids to clinical investigation.


Gelatin zymography and western blot analysis were applied in effusions from knees of the patients with septic (SA—10 patients), rheumatic (RA—10 patients) and osteoarthritis (OA—10 patients).


Zymographic analysis revealed that all samples contained latent MMP-2 activity, albeit activated MMP-2 appeared in more of the septic than aseptic effusions. MMP-9 was not detected in osteoarthritic synovial fluid samples. Only trace amounts of MMP-9 activity were detected in 4 of 10 patients with RA, whereas higher MMP-9 levels were evident in all samples from SA (P = 0.0241). In immunoblotting assays, samples from SA showed significantly higher levels of MMP-9 compared with samples from RA (P = 0.0052), confirming zymographic results. Although no significant difference in TIMP-1 levels was observed, the estimated MMP-9/TIMP-1 ratio of septic effusions was significantly higher compared with aseptic ones (P = 0.0029).


The data presented suggest enhanced expression and activation of MMP-9 in septic native knee arthritis compared with aseptic. The presence of high levels of MMP-9 with concomitantly increased MMP-9/TIMP-1 ratio and activated gelatinases in effusions, independent of neutrophilic counts, may be indicative for infection.


Septic Knee Arthritis MMP-9 TIMP-1 Gelatinases 

Supplementary material

167_2011_1676_MOESM1_ESM.doc (42 kb)
Supplementary material 1 (DOC 41 kb)


  1. 1.
    Aasen AO, Wang JE (2006) Mediator responses in surgical infections. Surg Infect 7(suppl 2):S3–S4Google Scholar
  2. 2.
    Arican M, Coughlan AR, Clegg PD, Carter SD (2000) MMP-2 and -9 activity in bovine synovial fluids. J Vet Med. A Physiol Pathol Clin Med 47:449–456PubMedCrossRefGoogle Scholar
  3. 3.
    Calander AM, Starckx S, Opdenakker G et al (2006) Matrix metalloproteinase-9 (gelatinase B) deficiency leads to increased severity of Staphylococcus aureus-triggered septic arthritis. Microbes Infect 8:1434–1439PubMedCrossRefGoogle Scholar
  4. 4.
    Chu S, Yang S, Lue K et al (2004) Clinical significance of gelatinases in septic arthritis of native and replaced knees. Clin Orthop Rel Res 427:179–183CrossRefGoogle Scholar
  5. 5.
    Elkington PTG, O’Kane CM, Friedland JS (2005) The paradox of MMPs in infectious disease. Clin Exp Immunol 142:12–20PubMedCrossRefGoogle Scholar
  6. 6.
    Fernando de Souza L, Jardim FR, Sauter IP et al (2008) High glucose increases RAW 264.7 macrophages activation by lipoteichoic acid from S. aureus. Clin Chim Acta 398:130–133CrossRefGoogle Scholar
  7. 7.
    Fietz S, Einspanier R, Hoppner S et al (2008) Determination of MMP-2 and -9 activities in synovial fluid of horses with osteoarthritic and arthritic joint diseases using gelatin zymography and immunocapture activity assays. Equine Vet J 40:266–271PubMedCrossRefGoogle Scholar
  8. 8.
    Firestein GS, Budd RC, Harris ED Jr et al (2008) Kelley’s textbook of rheumatology, 8th ed. Saunders, Philadelphia, pp 1–1902Google Scholar
  9. 9.
    Francoz D, Desrochers A, Simard N et al (2008) Relative expression of matrix metalloproteinase-2 and -9 in synovial fluid from healthy calves and calves with experimentally induced septic arthritis. Am J Vet Res 69:1022–1028PubMedCrossRefGoogle Scholar
  10. 10.
    Gehre F, Leib SL, Grandgirard D et al (2008) Essential role of choline for pneumococcal virulence in an experimental model of meningitis. J Intern Med 264:143–154PubMedCrossRefGoogle Scholar
  11. 11.
    Goldberg GI, Strongin A, Collier IE et al (1992) Interaction of 92-kDa type IV collagenase with TIMPs prevents dimerization, complex formation with interstitial collagenase, and activation of the proenzyme with stromelysin. J Biol Chem 267:4583–4591PubMedGoogle Scholar
  12. 12.
    Gupta MN, Sturrock RD, Field M (2001) A prospective 2-year study of 75 patients with adult-onset septic arthritis. Rheumatol (Oxford) 40:24–30CrossRefGoogle Scholar
  13. 13.
    Jalian HR, Liu PT, Kanchanapoomi M et al (2008) All-trans retinoic acid shifts Propionibacterium acnes-induced matrix degradation expression profile toward matrix preservation in human monocytes. J Invest Dermatol 128:2777–2782PubMedCrossRefGoogle Scholar
  14. 14.
    Jobin MC, Gottschalk M, Grenier D (2006) Upregulation of PGE2 and MMP-9 production by human macrophage-like cells: synergistic effect of capsular material and cell wall from Streptococcus suis. Microb Pathog 40:29–34PubMedCrossRefGoogle Scholar
  15. 15.
    Jotwani R, Eswaran SV, Moonga S, Cutler CW (2010) MMP-9/TIMP-1 imbalance induced in human dendritic cells by Porphyromonas gingivalis. FEMS Immunol Med Microbiol 58:314–321PubMedGoogle Scholar
  16. 16.
    Kanangat S, Postlethwaite A, Hasty K et al (2006) Induction of multiple MMPs in human dermal and synovial fibroblasts by S. aureus: implications in the pathogenesis of septic arthritis and other soft tissue infections. Arthr Res Ther 8:R176CrossRefGoogle Scholar
  17. 17.
    Kleiner DE, Stetler-Stevenson WG (1994) Quantitative zymography: detection of picogram quantities of gelatinases. Anal Biochem 218:325–329PubMedCrossRefGoogle Scholar
  18. 18.
    Lorente L, Martin MM, Labarta L et al (2009) Matrix metalloproteinase-9, -10, and tissue inhibitor of matrix metalloproteinases-1 blood levels as biomarkers of severity and mortality in sepsis. Crit Care 13:1006CrossRefGoogle Scholar
  19. 19.
    Ma L, Cranney A, Holroyd-Leduc JM (2009) Acute monoarthritis: what is the cause of my patient’s painful swollen joint? CMAJ 180:59–65PubMedCrossRefGoogle Scholar
  20. 20.
    Makowski GS, Ramsby ML (2003) Zymographic analysis of latent and activated forms of MMP-2 and -9 in synovial fluid: correlation to polymorphonuclear leukocyte infiltration and in response to infection. Clin Chim Acta 329:77–81PubMedCrossRefGoogle Scholar
  21. 21.
    Margaretten ME, Kohlwes J, Moore D, Bent S (2007) Does this patient have septic arthritis? JAMA 297:1478–1488PubMedCrossRefGoogle Scholar
  22. 22.
    Oggioni MR, Memmi G, Maggi T et al (2003) Pneumococcal zinc metalloproteinase ZmpC cleaves human MMP-9 and is a virulence factor in experimental pneumonia. Mol Microbiol 49:795–805PubMedCrossRefGoogle Scholar
  23. 23.
    Okada Y, Naka K, Kawamura K et al (1995) Localization of MMP-9 in osteoclasts: implications for bone resorption. Lab Invest 72:311–322PubMedGoogle Scholar
  24. 24.
    Punzi L, Oliviero F, Plebani M (2005) New biochemical insights into the pathogenesis of osteoarthritis and the role of laboratory investigations in clinical assessment. Crit Rev Clin Lab Sci 42:279–309PubMedCrossRefGoogle Scholar
  25. 25.
    Schaaf B, Liebau C, Kurowski V et al (2008) Hospital acquired pneumonia with high-risk bacteria is associated with increased pulmonary matrix metalloproteinase activity. BMC Pulm Med 8:12PubMedCrossRefGoogle Scholar
  26. 26.
    Sellner J, Leib SL (2006) In bacterial meningitis cortical brain damage is associated with changes in parenchymal MMP-9/TIMP-1 ratio and increased collagen type IV degradation. Neurobiol Dis 21:647–656PubMedCrossRefGoogle Scholar
  27. 27.
    Snoek-van Beurden PA, von den Hoff JW (2005) Zymographic techniques for the analysis of MMPs and their inhibitors. Biotechniques 38:73–83PubMedCrossRefGoogle Scholar
  28. 28.
    Taggart CC, Greene CM, Carroll TP et al (2005) Elastolytic proteases: inflammation resolution and dysregulation in chronic infective lung disease. Am J Respir Crit Care Med 171:1070–1076PubMedCrossRefGoogle Scholar
  29. 29.
    Tamura F, Nakagawa R, Akuta T et al (2004) Proapoptotic effect of proteolytic activation of MMPs by Streptococcus pyogenes thiol proteinase. Infect Immun 72:4836–4847PubMedCrossRefGoogle Scholar
  30. 30.
    Tanabe S, Bodet C, Grenier D (2007) Peptostreptococcus micros cell wall elicits a pro-inflammatory response in human macrophages. J Endotoxin Res 13:219–226PubMedCrossRefGoogle Scholar
  31. 31.
    Triebel S, Blaser J, Reinke H, Tschesche H (1992) A 25 kDa alpha 2-microglobulin-related protein is a component of the 125 kDa form of human gelatinase. FEBS Lett 314:386–388PubMedCrossRefGoogle Scholar
  32. 32.
    Van Doren SR, Wei S, Gao G et al (2008) Inactivation of N-TIMP-1 by N-terminal acetylation when expressed in bacteria. Biopolymers 89:960–968PubMedCrossRefGoogle Scholar
  33. 33.
    Visser S, Tupper J (2009) Septic until proven otherwise. Approach to and treatment of the septic joint in adult patients. Can Fam Phys 55:374–375Google Scholar
  34. 34.
    Wang JE, Pettersen S, Stuestol JF et al (2004) Peptidoglycan of S. aureus causes increased levels of MMPs in the rat. Shock 22:376–379PubMedCrossRefGoogle Scholar
  35. 35.
    Wang YY, Myhre AE, Pettersen SJ et al (2005) Peptidoglycan of S. aureus induces enhanced levels of MMP-9 in human blood originating from neutrophils. Shock 24:214–218PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2011

Authors and Affiliations

  • Vasileios Christos Fotopoulos
    • 1
    • 2
  • Athina Tzinia
    • 3
  • Matthew Tzurbakis
    • 2
  • Vasiliki Kalfakakou
    • 1
  • Stamatina Levidiotou-Stefanou
    • 1
  • Anastasios Georgoulis
    • 1
  1. 1.Department of MedicineUniversity of IoanninaIoanninaGreece
  2. 2.1st Orthopaedic Department“Evangelismos” General Hospital of AthensAthensGreece
  3. 3.Institute of BiologyNational Centre for Scientific Research “Demokritos”AthensGreece

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