Browsing by Subject "periodontal diseases"

Now showing 1 - 2 of 2
  • Thumbnail Image
    Item
    THE EFFECTS OF TOBACCO TREATED PORPHYROMONAS GINGIVALIS ON HUMAN EPITHELIAL CELLS
    (Office of the Vice Chancellor for Research, 2012-04-13) Tursunova, Roziya H.; Al-Shibani, Nouf; Windsor, L. Jack; Gregory, Richard L.
    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).
  • Thumbnail Image
    Item
    Nicotine-Treated Fusobacterium nucleatum Binding to Collagen, Fibrinogen, and Fibronectin
    (Office of the Vice Chancellor for Research, 2016-04-08) Beshay, Y.S.; Gregory, R.L.
    Fusobacterium nucleatum, a gram-negative anaerobic bacterium found in dental plaque, causes periodontal diseases. Smoking is one of the risk factors that can increase periodontal problems and atherosclerosis. Atherosclerosis is initiated by oral bacteria (i.e., F. nucleatum) binding to surface proteins of endothelial cells, such as collagen, fibrinogen, and fibronectin. The main objective for this study was to test the binding of F. nucleatum to collagen, fibrinogen, and fibronectin under the effect of different concentrations of nicotine. F. nucleatum was grown overnight in brain-heart infusion (BHI) supplemented with yeast extract and 5% vitamin-K/hemin. Biofilm was grown for 48 hours in 0, 0.25, 0.5, 1, and 2 mg/mL of nicotine. Then, the biofilm cells were labeled with biotin 3-sulfo-N-hydroxy-succinimide ester sodium salt and fixed with 10% formaldehyde. A binding assay was conducted by coating a high-binding 96-well microtiter plate with 1 μg/mL of collagen, fibrinogen, or fibronectin. The plate was incubated overnight and blocked with 1% Bovine Serum Albumin (BSA), followed by the biotinylated and nicotine-treated F. nucleatum cells. ExtrAvidin-Peroxidase and OPD Peroxidase Substrate was used to visualize the binding. Optical density (OD) was measured with a spectrophotometer at 490 nm. Collagen, fibrinogen, and fibronectin binding assays demonstrated significantly higher absorbance with 2 mg/mL nicotine-treated F. nucleatum cells compared to untreated cells. The results indicated that an increase in nicotine concentration leads to an increase in F. nucleatum binding to collagen, fibrinogen, and fibronectin. This means that smokers may have an increased risk for atherosclerosis. Supported by Life-Health and Sciences Internship (LHSI).