THE EFFECTS OF TOBACCO TREATED PORPHYROMONAS GINGIVALIS ON HUMAN EPITHELIAL CELLS

Abstract

Bacteria and tobacco are risk factors for periodontal diseases. Bacteria-host interactions play a critical role in disease development and progression. The effects of tobacco-treated bacteria such as Porphyromonas gingivalis on epithelial cells have not yet been examined. Therefore, P. gingivalis were treated with different tobacco products (nicotine, cigarette smoke conden-sate (CSC), and dissolvable smokeless tobacco (DST) strips) to determine the effects that they have on epithelial cells. P. gingivalis were grown with or without the products for 24 hours at 37◦C. The cells were separated from the supernatant, washed with 0.9% NaCl and incubated at 60◦C to kill the bacte-ria. Protein assays was performed to determine the protein concentration in the cell pellets and supernatants. Lactate dehydrogenase (LDH) assays are being used to measure the cytotoxicity of the cells and supernatants on epi-thelial cells in a dose dependent manner. Non-toxic amounts of the cell pel-lets and supernatants will be used to treat epithelial cells for 72 hours and the media analyzed by cytokine/growth factor protein arrays. The protein assays showed that CSC and nicotine treated P. gingivalis cells had less pro-tein than the others. The total protein in the supernatant for the CSC treated bacteria was less compared to others. The protein data suggests that CSC and nicotine affect protein expression in and by the P. gingivalis cells. To-bacco-treated bacteria are hypothesized to increase the expression of pro-inflammatory cytokines/growth factors by the epithelial cells, thereby con-tributing to the inflammation seen in periodontal diseases. This research was funded by Indiana University-Purdue University Indianapolis, Multidisci-plinary Undergraduate Research Institute (MURI).