Effects of Nicotine on Streptococcus gordonii Growth, Biofilm Formation, and Cell Aggregation

Abstract

ABSTRACTStreptococcus gordoniiis a commensal species of human oral flora. It initiates dental biofilm formation and provides binding sites for later colonizers to attach to and generate mature biofilm. Smoking is the second highest risk factor for periodontal disease, and cigarette smoke extract has been reported to facilitatePorphyromonas gingivalis-S. gordoniidual-species biofilm formation. Our hypothesis is that nicotine, one of the most important and active components of tobacco, stimulatesS. gordoniimultiplication and aggregation. In the present study,S. gordoniiplanktonic cell growth (kinetic absorbance and CFU), biofilm formation (crystal violet stain and confocal laser scanning microscopy [CLSM]), aggregation with/without sucrose, and 11 genes that encode binding proteins or regulators of gene expression were investigated. Results demonstrated planktonic cell growth was stimulated by 1 to 4 mg/ml nicotine treatment. Biofilm formation was increased at 0.5 to 4 mg/ml nicotine. CLSM indicated bacterial cell mass was increased by 2 and 4 mg/ml nicotine, but biofilm extracellular polysaccharide was not significantly affected by nicotine. Cell aggregation was upregulated by 4, 8, and 16 mg/ml nicotine with sucrose and by 16 mg/ml nicotine without sucrose. Quantitative reverse transcriptase PCR indicatedS. gordoniiabpA,scaA,ccpA, andsrtAwere upregulated in planktonic cells by 2 mg/ml nicotine. In conclusion, nicotine stimulatesS. gordoniiplanktonic cell growth, biofilm formation, aggregation, and gene expression of binding proteins. Those effects may promote later pathogen attachment to tooth surfaces, the accumulation of tooth calculus, and the development of periodontal disease in cigarette smokers.