Nicotine modulation of in vitro human gingival fibroblast β1 integrin expression

Harold B. Snyder, Gretchen B Caughman, Jill Lewis, Michael A. Billman, George Schuster

Research output: Contribution to journalArticle

24 Citations (Scopus)

Abstract

Background: Smoking is clearly a high-risk behavior for the development of periodontal disease, and nicotine is the major vasoactive component in tobacco. Integrins are a large family of homologous transmembrane adhesion proteins serving as the principal receptors on animal cells to bind and communicate with many extracellular matrix proteins such as collagen, fibronectin, and laminin. This study's aim was to evaluate nicotine's effects on β1 integrin expression as a function of either 1) the generalized effects on RNA/protein synthesis or 2) as a specific modulation of β1 integrin synthesis. Methods: Pooled human gingival fibroblasts (HGFs) were cultured in nicotine concentrations commonly seen in smokers (0.025 to 0.8 μM), and relative incorporation of [35S]methionine into newly synthesized protein or [3H]uridine into newly synthesized RNA was measured. Cultures were harvested at various times, and duplicate cell aliquots were homogenized and fractionated to obtain cell membrane-enriched preparations or solubilized to obtain whole cell lysates. Radiolabeled RNA and proteins were quantitated by trichloroacetic acid (TCA) precipitation and liquid scintillation spectrometry. β1 integrin subunits were detected by SDS-PAGE and Western blotting, and the relative intensities of reactive bands were quantitated by scanning densitometry. Results: After 17 hours of exposure, 0.4 and 0.8 μM nicotine resulted in a dose-dependent increase in β1 integrin in whole cell lysates, and a decrease in β1 integrin in the corresponding membrane-enriched fractions. There was also a statistically significant decrease (P ≤ 0.05) in radiolabeled proteins in culture. Although there appeared to be a mild, generalized reduction in radiolabeled RNA in nicotine-treated cultures compared to controls, a 1-way analysis of variance showed no statistical significance between values. Conclusions: Our results suggest that nicotine may induce an altered compartmentalization process in which β1 integrin molecules are produced, but are not appropriately transferred to the membrane. Nicotine effects on cellular protein synthesis and its modulation of β1 integrin expression may impair gingival fibroblast ability to adhere to and communicate with one another and with the extracellular matrix, which could impair wound healing and/or exacerbate periodontal disease.

Original languageEnglish (US)
Pages (from-to)505-510
Number of pages6
JournalJournal of Periodontology
Volume73
Issue number5
DOIs
StatePublished - Jun 3 2002
Externally publishedYes

Fingerprint

Nicotine
Integrins
Fibroblasts
RNA
Proteins
Periodontal Diseases
Trichloroacetic Acid
Membranes
In Vitro Techniques
Densitometry
Extracellular Matrix Proteins
Uridine
Laminin
Risk-Taking
Fibronectins
Methionine
Wound Healing
Tobacco
Extracellular Matrix
Polyacrylamide Gel Electrophoresis

Keywords

  • Fibroblasts, gingival
  • Integrins
  • Nicotine/adverse effects
  • Protein synthesis
  • Smoking/adverse effects

ASJC Scopus subject areas

  • Dentistry(all)

Cite this

Nicotine modulation of in vitro human gingival fibroblast β1 integrin expression. / Snyder, Harold B.; Caughman, Gretchen B; Lewis, Jill; Billman, Michael A.; Schuster, George.

In: Journal of Periodontology, Vol. 73, No. 5, 03.06.2002, p. 505-510.

Research output: Contribution to journalArticle

Snyder, Harold B. ; Caughman, Gretchen B ; Lewis, Jill ; Billman, Michael A. ; Schuster, George. / Nicotine modulation of in vitro human gingival fibroblast β1 integrin expression. In: Journal of Periodontology. 2002 ; Vol. 73, No. 5. pp. 505-510.
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AU - Schuster, George

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N2 - Background: Smoking is clearly a high-risk behavior for the development of periodontal disease, and nicotine is the major vasoactive component in tobacco. Integrins are a large family of homologous transmembrane adhesion proteins serving as the principal receptors on animal cells to bind and communicate with many extracellular matrix proteins such as collagen, fibronectin, and laminin. This study's aim was to evaluate nicotine's effects on β1 integrin expression as a function of either 1) the generalized effects on RNA/protein synthesis or 2) as a specific modulation of β1 integrin synthesis. Methods: Pooled human gingival fibroblasts (HGFs) were cultured in nicotine concentrations commonly seen in smokers (0.025 to 0.8 μM), and relative incorporation of [35S]methionine into newly synthesized protein or [3H]uridine into newly synthesized RNA was measured. Cultures were harvested at various times, and duplicate cell aliquots were homogenized and fractionated to obtain cell membrane-enriched preparations or solubilized to obtain whole cell lysates. Radiolabeled RNA and proteins were quantitated by trichloroacetic acid (TCA) precipitation and liquid scintillation spectrometry. β1 integrin subunits were detected by SDS-PAGE and Western blotting, and the relative intensities of reactive bands were quantitated by scanning densitometry. Results: After 17 hours of exposure, 0.4 and 0.8 μM nicotine resulted in a dose-dependent increase in β1 integrin in whole cell lysates, and a decrease in β1 integrin in the corresponding membrane-enriched fractions. There was also a statistically significant decrease (P ≤ 0.05) in radiolabeled proteins in culture. Although there appeared to be a mild, generalized reduction in radiolabeled RNA in nicotine-treated cultures compared to controls, a 1-way analysis of variance showed no statistical significance between values. Conclusions: Our results suggest that nicotine may induce an altered compartmentalization process in which β1 integrin molecules are produced, but are not appropriately transferred to the membrane. Nicotine effects on cellular protein synthesis and its modulation of β1 integrin expression may impair gingival fibroblast ability to adhere to and communicate with one another and with the extracellular matrix, which could impair wound healing and/or exacerbate periodontal disease.

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