Binding, uptake, and release of nicotine by human gingival fibroblasts.
Previous studies of the effects of nicotine on fibroblasts have reported an altered morphology and attachment of fibroblasts to substrates and disturbances in protein synthesis and secretion. This altered functional and attachment response may be associated with changes in the cell membrane resulting from binding of the nicotine, or to disturbances in cell metabolism as a result of high intracellular levels of nicotine. The purpose of the present study, therefore, was to 1) determine whether gingival fibroblasts bound nicotine and if any binding observed was specific or non-specific in nature; 2) determine whether gingival fibroblasts internalized nicotine, and if so, at what rate; 3) determine whether gingival fibroblasts also released nicotine back into the extracellular environment; and 4) if gingival fibroblasts release nicotine intact or as a metabolite. Cultures of gingival fibroblasts were prepared from gingival connective tissue biopsies. Binding was evaluated at 4 degrees C using a mixture of 3H-nicotine and unlabeled nicotine. Specific binding was calculated as the difference between 3H-nicotine bound in the presence and absence of unlabeled nicotine. The cells bound 1.44 (+/- 0.42) pmols/10(6) cells in the presence of unlabeled nicotine and 1.66 (+/- 0.55) pmols/10(6) cells in the absence of unlabeled nicotine. The difference was not significant. Uptake of nicotine was measured at 37 degrees C after treating cells with 3H-nicotine for time periods up to 4 hours. Uptake in pmols/10(6) cells was 4.90 (+/- 0.34) at 15 minutes, 8.30 (+/- 0.75) at 30 minutes, 12.28 (+/- 2.62) at 1 hour and 26.31 (+/- 1.15) at 4 hours.(ABSTRACT TRUNCATED AT 250 WORDS)
Hanes
P J
PJ
Department of Periodontics, School of Dentistry, Medical College of Georgia, Augusta.
Schuster
G S
GS
Lubas
S
S
D000328:Adult; D002478:Cells, Cultured; D002845:Chromatography; D005260:Female; D005347:Fibroblasts / Q000378:metabolism; D005881:Gingiva / Q000378:metabolism / Q000473:pathology*; D006801:Humans; D008297:Male; D009538:Nicotine / Q000378:metabolism / Q000493:pharmacokinetics*; D010518:Periodontitis / Q000378:metabolism / Q000473:pathology*; D013997:Time Factors; D014316:Tritium
D014316:Tritium; D009538:Nicotine