BACKGROUND: Integrins are a family of transmembrane cell surface glycoproteins, and those with the beta 1-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 beta 1 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 microM. Human gingival fibroblasts (HFG) were grown for 24 hours in each concentration and fluorescein-labeled with a mouse monoclonal anti-human beta 1 antibody and secondarily incubated with a urease-labeled anti-mouse IgG antibody. After a final wash, the cells were incubated with urea/bromcresol blue substrate for 15 minutes at 37 degrees C and measured in a microplate reader at 570 nm. RESULTS: The integrin beta 1-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 microM versus controls (P < 0.05). CONCLUSIONS: Nicotine concentrations of 0.2 and 0.4 microM significantly decrease beta 1 integrin expression in human gingival fibroblasts that may affect cell-cell and cell-substratum adhesion during wound healing.
BACKGROUND: Integrins are a family of transmembrane cell surface glycoproteins, and those with the beta 1-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 beta 1 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 microM. Human gingival fibroblasts (HFG) were grown for 24 hours in each concentration and fluorescein-labeled with a mouse monoclonal anti-humanbeta 1 antibody and secondarily incubated with a urease-labeled anti-mouse IgG antibody. After a final wash, the cells were incubated with urea/bromcresol blue substrate for 15 minutes at 37 degrees C and measured in a microplate reader at 570 nm. RESULTS: The integrin beta 1-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 microM versus controls (P < 0.05). CONCLUSIONS:Nicotine concentrations of 0.2 and 0.4 microM significantly decrease beta 1 integrin expression in human gingival fibroblasts that may affect cell-cell and cell-substratum adhesion during wound healing.
Authors: Marco Tatullo; Stefano Gentile; Francesco Paduano; Luigi Santacroce; Massimo Marrelli Journal: Medicine (Baltimore) Date: 2016-12 Impact factor: 1.817