title: Measurement of matrix metalloproteinase 9-mediated collagen type III degradation fragment as a marker of skin fibrosis.
abstract: BACKGROUND
  The current study utilized a Bleomycin-induced model of skin fibrosis to investigate the neo-epitope CO3-610 (KNGETGPQGP), a fragment of collagen III released during matrix metalloproteinase-9 (MMP9) degradation of the protein, we have previously described as a novel biomarker for liver fibrosis. The aim was to investigate CO3-610 levels in another well characterised model of fibrosis, to better describe the biomarker in relation to additional fibrotic pathologies.
  
  
  METHODS
  Skin fibrosis was induced by daily injections of Bleomycin to a total of 52 female C3 H mice, while control mice (n = 28) were treated with phosphate buffered saline (PBS), for 2, 4, 6 or 8 weeks. Skin fibrosis was evaluated using Visiopharm software on Sirius-red stained skin sections. Urine ELISA assays and creatinine corrections were performed to measure CO3-610 levels.
  
  
  RESULTS
  CO3-610 levels were significantly higher in Bleomycin-treated vs. PBS-treated mice at each time point of termination. The mean increases were: 59.2%, P < 0.0008, at 2 weeks; 113.5%, P < 0.001, at 4 weeks; 136.8%, P < 0.0001 at 6 weeks; 157.2%, P < 0.0001 at 8 weeks). PBS-treated mice showed a non-significant increase in CO3-610 levels (mean increase for weeks 2-8 = 1.7%, P = 0.789) CO3-610 levels assayed in urine were statistically significantly correlated with Western blot analysis showing increased skin fibrosis (P < 0.0001, r = 0.65).
  
  
  CONCLUSIONS
  Increased levels in mouse urine of the MMP-9 mediated collagen III degradation fragment CO3-610 were correlated with skin fibrosis progression, suggesting that CO3-610 may be a potential positive biomarker to study the pathogenesis of skin fibrosis in mice.
authors: Assay Development, Nordic Bioscience, Herlev Hovedgade 207, DK-2730, Copenhagen, Denmark. eva@nordicbioscience.com
authors: Efstathios
authors: E
authors: Vassiliadis
authors: Sanne Skovgård
authors: SS
authors: Veidal
authors: Natasha
authors: N
authors: Barascuk
authors: Jhinuk Basu
authors: JB
authors: Mullick
authors: Rikke Elgaard
authors: RE
authors: Clausen
authors: Lise
authors: L
authors: Larsen
authors: Henrik
authors: H
authors: Simonsen
authors: Dorthe Vang
authors: DV
authors: Larsen
authors: Anne-Christine
authors: AC
authors: Bay-Jensen
authors: Toni
authors: T
authors: Segovia-Silvestre
authors: Diana Julie
authors: DJ
authors: Leeming
authors: Morten A
authors: MA
authors: Karsdal
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000652:urine; D001761:Bleomycin; D024061:Collagen Type III / Q000652:urine*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D000939:Epitopes / Q000652:urine*; D005109:Extracellular Matrix; D005260:Female; D005355:Fibrosis / Q000139:chemically induced / Q000378:metabolism* / Q000473:pathology; D020780:Matrix Metalloproteinase 9 / Q000502:physiology; D051379:Mice; D008809:Mice, Inbred C3H; D012871:Skin Diseases / Q000139:chemically induced / Q000378:metabolism* / Q000473:pathology
chemical_list: D015415:Biomarkers; D024061:Collagen Type III; D000939:Epitopes; D001761:Bleomycin; D020780:Matrix Metalloproteinase 9; C501830:Mmp9 protein, mouse