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Its Stimulating and Inhibitory Effects on Oral Microorganisms Aubrey E

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Its Stimulating and Inhibitory Effects on Oral Microorganisms Aubrey E NICOTINE: ITS STIMULATING AND INHIBITORY EFFECTS ON ORAL MICROORGANISMS AUBREY E. DUBOIS*1, ZACHARY C. BENNETT*1, UMARA KHALID2, ARIBA KHALID1, REED A. MEECE1, GABRIELLE J. DIFIORE3, AND RICHARD L. GREGORY4 1DEPARTMENT OF BIOLOGY, SCHOOL OF SCIENCE, PURDUE UNIVERSITY, INDIANAPOLIS, IN; 2DEPARTMENT OF PSYCHOLOGY, SCHOOL OF SCIENCE, PURDUE UNIVERSITY, INDIANAPOLIS, IN; 3DEPARTMENT OF KINESIOLOGY, SCHOOL OF PHYSICAL EDUCATION AND TOURISM MANAGEMENT, INDIANA UNIVERSITY, INDIANAPOLIS, IN; 4DEPARTMENT OF ORAL BIOLOGY, SCHOOL OF DENTISTRY, INDIANA UNIVERSITY, INDIANAPOLIS, IN *A.E.D. AND Z.C.B. ARE JOINT FIRST AUTHORS OF THIS WORK. MANUSCRIPT RECEIVED 15 SEPTEMBER, 2014; ACCEPTED 30 OCTOBER, 2014 Copyright 2014, Fine Focus all rights reserved 64 • FINE FOCUS, VOL. 1 ABSTRACT Tobacco users are much more susceptible to dental caries and periodontal diseases than non- tobacco users. Research suggests that this increased susceptibility may be due in part to nicotine, a primary active component of tobacco. Five bacterial species and one yeast species commonly found in the human oral cavity, Lactobacillus casei, Actinomyces viscosus, Actinomyces naeslundii, Rothia dentocariosa, Enterococcus faecalis, and Candida albicans respectively, were utilized to investigate if any correlation existed between CORRESPONDING exposure to various concentrations of nicotine ranging from 0 to 32 mg/ml and the growth of AUTHOR each microorganism. The minimum inhibitory concentration (MIC), minimum bioflm inhibitory Richard L. Gregory concentration (MBIC), and planktonic growth were [email protected] measured. The MIC was determined to be 16 mg/ ml for all organisms except E. faecalis, which had an MIC of 32 mg/ml. Nicotine had a varying effect on planktonic growth across the different species. KEYWORDS A distinct upward trend in bioflm formation was found in A. viscosus, L. casei, E. faecalis, • dental caries and C. albicans through 8 mg/ml. Nicotine also • periodontal disease enhanced R. dentocariosa bioflm formation in • bioflm all concentrations through 8 mg/ml but was most • nicotine enhanced at 1 mg/ml. Alternatively, A. naeslundii • tobacco exhibited a complete downward trend through 32 • Lactobacillus casei mg/ml. The MBIC was found to be 16 mg/ml in all • Actinomyces viscosus organisms studied. These fndings further support • Actinomyces naeslundii research suggesting that the increased susceptibility • Rothia dentocariosa to oral health diseases experienced by tobacco • Enterococcus faecalis users may be caused in part by an upregulation in • Candida albicans bioflm formation of these oral pathogens. INTRODUCTION Periodontal disease is a condition that in which the gums pull away from the causes infammation of the gums including teeth and form pockets where dental plaque diseases such as gingivitis, in which begins to grow. This infection can lead to gums become swollen and bleed easily. both severe tissue and bone damage due Periodontitis, another form of periodontal to the increased exposure to previously disease, is an advanced form of gingivitis protected areas. Periodontal disease is also BACTERIAL HOST INTERACTIONS • 65 closely linked with dental caries. The lacks adequate strength to adhere to tooth occurrence of both periodontal disease enamel, however, when co-cultured with and dental caries can be associated with Streptococcus mutans, L. casei gains the several factors. Some of the most common ability to synthesize glucans thus improving risk factors include smoking, diabetes, tooth enamel adhesion. Because of this, L. autoimmune disorders, genetics, and obesity casei along with S. mutans have been found (14). The use of tobacco products remains to be the most prevalent bacteria leading to the largest risk factor for periodontal dental caries (19). disease. Environmental tobacco smoke has been associated with the increased ACTINOMYCES VISCOSUS likelihood of caries formation in children Actinomyces viscosus is a facultative (2). Recent studies in this laboratory anaerobic, Gram-positive bacterium that is demonstrated increased bioflm formation often isolated from the oral cavity, especially of S. mutans with increasing nicotine from subgingival plaque and along the teeth concentrations (20). The average nicotine and gums. A. viscosus is rod-shaped and content in a single cigarette has been found flamentous. Distinct characteristics of the in amounts up to 2 mg (12). A recent study flamentous shape include branching with tested over 40 smokeless tobacco products, swollen, rounded, or clavate ends (7). A. such as snuff and chew, and it was viscosus is a severely cariogenic bacterium discovered that they contained anywhere that acts as an agent in promoting and from 3.6-25.3 mg of nicotine per gram of initializing carious lesions (15). Studies have smokeless tobacco (33). Nicotine contents shown that the bacterium has adhesins of cigars have been found to vary greatly, which allow it to bind to complementary ranging from 5.9 mg per cigar to 335.2 mg receptors on a substrate. A. viscosus binds to per cigar (18). Studies have demonstrated salivary acidic proline-rich proteins (PRPs) that when smoking cigarettes, an average adsorbed onto the apatitic surfaces of the of approximately 1 mg of nicotine per tooth. Cryptic segments exposed in adsorbed cigarette is absorbed by the body, however, molecules are identifed by the adhesins, the amount of nicotine within oral bioflm which allow A. viscosus to effciently attach of smokers has not yet been explored (5). to teeth while planktonically suspended in This study used the following opportunistic saliva (1). pathogens, commonly found in the human oral cavity, to investigate possible effects on ACTINOMYCES NAESLUNDII growth and bioflm formation. Actinomyces naeslundii is a Gram-positive bacterium characterized by its rod shape LACTOBACILLUS CASEI and prominent fmbriae that allow it to Lactobacillus casei is a Gram-positive, bind to the tooth surface. A. naeslundii is facultative anaerobic bacterium. They are facultatively anaerobic, nonmotile, non- nonmotile and have a rod shape. Like other spore forming, and commonly grows in the strains of lactobacilli, L. casei produces a oral cavity. (6) A. naeslundii is also a major signifcant amount of lactic acid, allowing it component of dental plaque and it is known to remain viable under various pH levels. to cause dental caries, root canal infections, Poor diet, weak tooth enamel, previous and periodontal disease (27). It is one of the plaque buildup, and acidic conditions specifc species of the genus Actinomyces contribute to an optimal environment capable of causing the chronic bacterial for L. casei to thrive (22). L. casei alone infection, actinomycosis, 66 • FINE FOCUS, VOL. 1 in humans (8). A. naeslundii plays a role carbohydrates, and a number of amino in actinomycosis by initiating infections acids. It is also able to withstand extremely in carious teeth, as well as causing basic environments, high salt concentrations, infections in places where the mucosal desiccation, and the presence of many integrity of the tooth is compromised (32). antibiotics (17). These attributes allow These infections are the precursor of the E. faecalis to thrive in deep root canals. actinomycosis, however, the bacterium Additionally, scientists recently discovered itself exhibits very low pathogenic E. faecalis has the ability to suppress properties. These properties are enhanced lymphocyte function as well as produce by the presence of microorganisms such as lytic enzymes which aid in destruction of species of Prevotella, Staphylococcus, and gingival tissues (9). Streptococcus (35). CANDIDA ALBICANS ROTHIA DENTOCARIOSA Candida albicans is a ubiquitous Rothia dentocariosa is a facultative anaerobic, pleomorphic fungus that colonizes the Gram-positive, pleomorphic bacterium that gastrointestinal, epithelial, and mucosal is commonly found in either flamentous tissues of over 70% of the human or coccoid forms (24). It is common in population (26). Whether C. albicans takes the oral cavity, and has been associated on hyphal, pseudohyphal, or yeast form with many types of bacterial infections. is dependent upon environmental factors Although R. dentocariosa was frst isolated such as temperature, pH, and presence in dental caries, endocarditis is the most of serum. C. albicans is an opportunistic common infection caused by the organism. pathogen that is often the cause of infection Cases have also been reported associating in immunocompromised patients, such R. dentocariosa with intrauterine fetal death as those with HIV/AIDS or indwelling (16,25,28). R. dentocariosa is commonly medical devices. Imbalances in normal associated with dental caries and periodontal body fora due to antibiotic and steroid disease. Studies have demonstrated the use have also been shown to provoke the ability of R. dentocariosa to induce the overgrowth of Candida, causing candidiasis release of TNF-a by macrophages, thereby (21). Oral candidiasis, commonly known as eliciting infammatory responses within the thrush, can exacerbate already existing oral gingiva (36). diseases. C. albicans has also been found within dental caries and is a contributor ENTEROCOCCUS FAECALIS to periodontal disease (26). Many proteins Enterococcus faecalis is a facultative within saliva have been found to promote anaerobic, Gram-positive cocci bacterium adherence of C. albicans to gingival tissues that is typically found in pairs or short (10). Additionally, tissues already colonized chains. The most common locations
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