Background: Patients who smoke are at increased risk for chronic periodontitis (CP). Most studies suggest that the microbial flora in these patients is similar to that found in non-smoking CP patients. Thus, the increased risk for development of CP is not dependent on an altered microbial profile, but rather to some change in the host response to these periopathogens. There is evidence that human gingival fibroblasts (HGF) derived from diseased sites produce greater amounts of interleukin (IL)-6 and IL-8 in vitro than cells derived from healthy sites. This suggests that HGF subpopulations may be selected based upon the inflammatory milieu in which they reside. The hypothesis to be tested was that the combination of nicotine and lipopolysaccharide (LPS) could regulate HGF inflammatory mediator production.

Methods: HGF cell cultures were established from explants derived from 10 patients with CP. HGF cell cultures were stimulated with 1mM, 1µM, or 1ηM nicotine ± Escherichia coli or Porphyromonas gingivalis LPS. At 12, 24, or 48-hour time points, the cells were counted and the supernatant was collected for subsequent IL-6 and IL-8 determination in an enzyme-linked immunosorbent assay.

Results: At the 24-hour time point, 1ηM nicotine stimulated IL-6 production compared to control (P = 0.02). E. coli LPS alone caused a 3- to 4-fold increase in IL-6 and IL-8 production, whereas P. gingivalis LPS did not augment IL-6 or IL-8. A synergistic effect upregulating IL-6 was observed with combined treatment of 1mM nicotine and E. coli LPS or P. gingivalis LPS at the 24-hour time point (P < 0.0005 and P = 0.002, respectively). Similar effects were seen when IL-8 production was evaluated following HGF stimulation with high doses of nicotine and E. coli LPS or P. gingivalis LPS.

Conclusions: These results demonstrate that nicotine by itself can stimulate HGF IL-6 and IL-8 production. Moreover, the combination of high doses of nicotine and either E. coli or P. gingivalis LPS can synergistically upregulate cytokine production. These findings support the hypothesis that a proinflammatory fibroblast phenotype may be elicited in an environment enriched with bacterial LPS and nicotine. J Periodontol 2001;72:1038-1044.