title: Nicotine and smokeless tobacco effects on gingival and peripheral blood mononuclear cells.
abstract: The pathogenesis of tobacco-related periodontal diseases is not well understood. The purpose of this study was therefore to investigate smokeless tobacco extract (ST) and nicotine effects on prostaglandin E2 (PGE2) and interleukin-1beta (IL-1beta) secretion by peripheral blood mononuclear cells (PBMC, consisting of monocytes and lymphocytes) and gingival mononuclear cells (GMC). Both peripheral blood and gingival tissue adjacent to the alveolar crest were taken from non-smoking adult periodontitis patients. Gingival tissue was treated with collagenase and deoxyribonuclease and GMC and PBMC were isolated by Ficoll-Hypaque centrifugation. GMC and PBMC (100,000 cells/200 microl) were cultured for 24 hours in supplemented RPMI 1640 alone (control), or in supplemented RPMI 1640 containing 1% ST, 100 microg/ml nicotine, 1 microg/ml Porphyromonas gingivalis LPS, or 1 microg/ml P. gingivalis LPS and either 100 microg/ml nicotine or 1% ST. Enzyme immunoassays were used to quantify PGE2 and IL-1beta. Treatments were compared by repeated measures ANOVA. 100 microg/ml nicotine (7-fold, p<0.02) and 1% ST (3.5-fold, p<0.004) significantly increased secretion of PGE2 by PBMC relative to control cultures. 100 microg/ml nicotine and 1% ST, however, had no effect on IL-1beta secretion by PBMC. Enhanced PGE2 secretion also was seen when PBMC were treated with P. gingivalis LPS+ 100 microg/ml nicotine relative to P. gingivalis LPS alone (p<0.007). In contrast, 100 microg/ml nicotine significantly downregulated IL-1beta secretion by GMC relative to medium alone (p<0.008) and had no effect on PGE2 secretion by GMC. These data indicate that while nicotine and ST can stimulate PBMC to secrete PGE2, they cannot activate further mononuclear cells extracted from gingiva, possibly due to maximal previous stimulation in the periodontitis lesion.
authors.affiliation: Department of Surgical Specialties, University of Nebraska Medical Center, College of Dentistry, Lincoln 68583, USA.
authors.forename: E
authors.initials: E
authors.lastname: Bernzweig
authors.forename: J B
authors.initials: JB
authors.lastname: Payne
authors.forename: R A
authors.initials: RA
authors.lastname: Reinhardt
authors.forename: J K
authors.initials: JK
authors.lastname: Dyer
authors.forename: K D
authors.initials: KD
authors.lastname: Patil
mesh_terms: D000704:Analysis of Variance; D002478:Cells, Cultured; D015232:Dinoprostone / Q000096:biosynthesis; D005881:Gingiva / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D007118:Immunoassay; D007375:Interleukin-1 / Q000096:biosynthesis; D007963:Leukocytes, Mononuclear / Q000187:drug effects* / Q000378:metabolism; D008875:Middle Aged; D009538:Nicotine / Q000009:adverse effects*; D010518:Periodontitis / Q000097:blood / Q000378:metabolism; D010947:Plants, Toxic; D014030:Tobacco, Smokeless / Q000009:adverse effects*
chemical_list: D007375:Interleukin-1; D009538:Nicotine; D015232:Dinoprostone