Effect of nicotine on fibroblast β1 integrin expression and distribution in vitro

G. W. Austin, M. F. Cuenin, S. D. Hokett, Mark E. Peacock, D. E. Sutherland, J. F. Erbland, M. A. Billman

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

Background: Integrins are a family of transmembrane cell surface glycoproteins, and those with the β1-subunit function in both cell-to-cell and cell-to-substrate adhesion. The purpose of this study was to determine nicotine's effect on the expression and distribution of the β1 integrin subunit on the human gingival fibroblast cell surface. Methods: Pure nicotine was diluted in medium to the following concentrations: 0 (control), 0.025, 0.05, 0.1, 0.2, and 0.4 μΜ. Human gingival fibroblasts (HFG) were grown for 24 hours in each concentration and fluorescein-labeled with a mouse monoclonal anti-human β1 antibody and secondarily incubated with a urease-labeled anti-mouse lgG antibody. After a final wash, the cells were incubated with urea/bromcresol blue substrate for 15 minutes at 37°C and measured in a microplate reader at 570 nm. Results: The integrin β1-subunit was detected on the HGF surface membrane by fluorescence labeling, and cell-enzyme-linked immunosorbent assay testing demonstrated its decreased expression with increasing nicotine concentrations that were statistically different at the concentrations of 0.2 and 0.4 μΜ versus controls (P<0.05). Conclusions: Nicotine concentrations of 0.2 and 0.4 μΜ significantly decrease β1 integrin expression in human gingival fibroblasts that may affect cell-cell and cell-substratum adhesion during wound healing.

Original languageEnglish (US)
Pages (from-to)438-444
Number of pages7
JournalJournal of Periodontology
Volume72
Issue number4
DOIs
StatePublished - Jun 21 2001
Externally publishedYes

Fingerprint

Nicotine
Integrins
Fibroblasts
Urease
In Vitro Techniques
Antibodies
Membrane Glycoproteins
Fluorescein
Cell Adhesion
Wound Healing
Urea
Fluorescence
Enzyme-Linked Immunosorbent Assay
Membranes

Keywords

  • Fibroblasts
  • Integrin beta-1 subunit
  • Nicotine/adverse effects

ASJC Scopus subject areas

  • Periodontics

Cite this

Austin, G. W., Cuenin, M. F., Hokett, S. D., Peacock, M. E., Sutherland, D. E., Erbland, J. F., & Billman, M. A. (2001). Effect of nicotine on fibroblast β1 integrin expression and distribution in vitro. Journal of Periodontology, 72(4), 438-444. https://doi.org/10.1902/jop.2001.72.4.438

Effect of nicotine on fibroblast β1 integrin expression and distribution in vitro. / Austin, G. W.; Cuenin, M. F.; Hokett, S. D.; Peacock, Mark E.; Sutherland, D. E.; Erbland, J. F.; Billman, M. A.

In: Journal of Periodontology, Vol. 72, No. 4, 21.06.2001, p. 438-444.

Research output: Contribution to journalArticle

Austin, GW, Cuenin, MF, Hokett, SD, Peacock, ME, Sutherland, DE, Erbland, JF & Billman, MA 2001, 'Effect of nicotine on fibroblast β1 integrin expression and distribution in vitro', Journal of Periodontology, vol. 72, no. 4, pp. 438-444. https://doi.org/10.1902/jop.2001.72.4.438
Austin, G. W. ; Cuenin, M. F. ; Hokett, S. D. ; Peacock, Mark E. ; Sutherland, D. E. ; Erbland, J. F. ; Billman, M. A. / Effect of nicotine on fibroblast β1 integrin expression and distribution in vitro. In: Journal of Periodontology. 2001 ; Vol. 72, No. 4. pp. 438-444.
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AU - Cuenin, M. F.

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AU - Sutherland, D. E.

AU - Erbland, J. F.

AU - Billman, M. A.

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AB - Background: Integrins are a family of transmembrane cell surface glycoproteins, and those with the β1-subunit function in both cell-to-cell and cell-to-substrate adhesion. The purpose of this study was to determine nicotine's effect on the expression and distribution of the β1 integrin subunit on the human gingival fibroblast cell surface. Methods: Pure nicotine was diluted in medium to the following concentrations: 0 (control), 0.025, 0.05, 0.1, 0.2, and 0.4 μΜ. Human gingival fibroblasts (HFG) were grown for 24 hours in each concentration and fluorescein-labeled with a mouse monoclonal anti-human β1 antibody and secondarily incubated with a urease-labeled anti-mouse lgG antibody. After a final wash, the cells were incubated with urea/bromcresol blue substrate for 15 minutes at 37°C and measured in a microplate reader at 570 nm. Results: The integrin β1-subunit was detected on the HGF surface membrane by fluorescence labeling, and cell-enzyme-linked immunosorbent assay testing demonstrated its decreased expression with increasing nicotine concentrations that were statistically different at the concentrations of 0.2 and 0.4 μΜ versus controls (P<0.05). Conclusions: Nicotine concentrations of 0.2 and 0.4 μΜ significantly decrease β1 integrin expression in human gingival fibroblasts that may affect cell-cell and cell-substratum adhesion during wound healing.

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