Effects of cigarette smoke condensate and nicotine on human gingival fibroblast-mediated collagen degradation

Weiping Zhang, Meixian Fang, Fengyu Song, L. Windsor

Research output: Contribution to journalArticle

25 Citations (Scopus)

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, aswell as on the protein andmRNAlevels of MMP-2, MMP-14, and TIMP-2. Conclusions: CSC may increase HGF-mediated collagen degradation by affecting membrane-associated MMPs and TIMPs.

Original languageEnglish (US)
Pages (from-to)1071-1079
Number of pages9
JournalJournal of Periodontology
Volume82
Issue number7
DOIs
StatePublished - Jul 2011

Fingerprint

Nicotine
Smoke
Tobacco Products
Collagen
Fibroblasts
Matrix Metalloproteinase Inhibitors
Matrix Metalloproteinase 2
Matrix Metalloproteinase 14
Tissue Inhibitor of Metalloproteinase-2
Tissue Inhibitor of Metalloproteinases
Periodontal Diseases
Conditioned Culture Medium
Messenger RNA
Aptitude
Membranes
Gelatin
Matrix Metalloproteinases
Reverse Transcription
Tobacco
Membrane Proteins

Keywords

  • Collagen
  • Matrix metalloproteinases
  • Nicotine
  • Periodontal diseases
  • Smoke

ASJC Scopus subject areas

  • Periodontics

Cite this

Effects of cigarette smoke condensate and nicotine on human gingival fibroblast-mediated collagen degradation. / Zhang, Weiping; Fang, Meixian; Song, Fengyu; Windsor, L.

In: Journal of Periodontology, Vol. 82, No. 7, 07.2011, p. 1071-1079.

Research output: Contribution to journalArticle

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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, aswell as on the protein andmRNAlevels of MMP-2, MMP-14, and TIMP-2. Conclusions: CSC may increase HGF-mediated collagen degradation by affecting membrane-associated MMPs and TIMPs.",
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