title: Effects of cigarette smoke condensate and nicotine on human gingival fibroblast-mediated collagen degradation.
abstract: BACKGROUND
  Members of the matrix metalloproteinase (MMP) family have been shown to be involved in periodontal disease. Risk factors for periodontal disease include tobacco smoking. Cigarette smoke condensate (CSC) is comprised of thousands of chemicals. Nicotine is one of the active components in tobacco. This study compares the effects of CSC and nicotine at the level in CSC on the collagen-degrading ability of human gingival fibroblasts (HGFs) and the expression of selected MMPs and tissue inhibitors of metalloproteinases (TIMPs).
  
  
  METHODS
  HGFs were seeded in six-well collagen-coated plates, exposed to 100 μg/mL (2.4 μg/mL nicotine) of CSC or 2.4 μg/mL nicotine for 3 days, and then collagen degradation was analyzed. After 3 days exposure to CSC or nicotine, the conditioned media from HGFs was collected and the membrane proteins were extracted for gelatin zymography and Western blot analyses. The mRNA levels of MMP-2, MMP-14, and TIMP-2 were measured by reverse transcription-polymerase chain reaction.
  
  
  RESULTS
  The CSC increased collagen degradation, and increased the levels of TIMP-2, MMP-14, and the active MMP-2 in the membrane extracts, and their mRNA levels. CSC also increased the level of active MMP-2 in the conditioned media. Nicotine at the level in CSC (2.4 μg/mL) had little influence on collagen degradation, as well as on the protein and mRNA levels of MMP-2, MMP-14, and TIMP-2.
  
  
  CONCLUSIONS
  CSC may increase HGF-mediated collagen degradation by affecting membrane-associated MMPs and TIMPs.
authors.affiliation: Department of Oral Biology, Indiana University School of Dentistry, Indianapolis, IN 46202, USA.
authors.forename: Weiping
authors.initials: W
authors.lastname: Zhang
authors.forename: Meixian
authors.initials: M
authors.lastname: Fang
authors.forename: Fengyu
authors.initials: F
authors.lastname: Song
authors.forename: L Jack
authors.initials: LJ
authors.lastname: Windsor
mesh_terms: D015153:Blotting, Western; D002478:Cells, Cultured; D024042:Collagen Type I / Q000032:analysis / Q000187:drug effects*; D045424:Complex Mixtures / Q000494:pharmacology; D017077:Culture Media, Conditioned; D016895:Culture Media, Serum-Free; D021983:Cyclophilins / Q000032:analysis / Q000187:drug effects; D004792:Enzyme Precursors / Q000032:analysis / Q000187:drug effects; D005347:Fibroblasts / Q000187:drug effects* / Q000378:metabolism; D018093:Gelatinases / Q000032:analysis / Q000187:drug effects; D005881:Gingiva / Q000166:cytology / Q000187:drug effects*; D006801:Humans; D053511:Matrix Metalloproteinase 14 / Q000032:analysis / Q000187:drug effects; D020778:Matrix Metalloproteinase 2 / Q000032:analysis / Q000187:drug effects; D053504:Matrix Metalloproteinases, Membrane-Associated / Q000032:analysis / Q000187:drug effects; D009538:Nicotine / Q000494:pharmacology*; D011480:Protease Inhibitors / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012906:Smoke; D019716:Tissue Inhibitor of Metalloproteinase-2 / Q000032:analysis / Q000187:drug effects; D019714:Tissue Inhibitor of Metalloproteinases / Q000032:analysis / Q000187:drug effects; D014026:Nicotiana
chemical_list: D024042:Collagen Type I; D045424:Complex Mixtures; D017077:Culture Media, Conditioned; D016895:Culture Media, Serum-Free; D004792:Enzyme Precursors; D011480:Protease Inhibitors; D012906:Smoke; C560823:TIMP2 protein, human; D019714:Tissue Inhibitor of Metalloproteinases; D019716:Tissue Inhibitor of Metalloproteinase-2; D009538:Nicotine; D018093:Gelatinases; D053504:Matrix Metalloproteinases, Membrane-Associated; C069860:progelatinase; C522361:MMP2 protein, human; D020778:Matrix Metalloproteinase 2; C507086:MMP14 protein, human; D053511:Matrix Metalloproteinase 14; D021983:Cyclophilins