The aim of this study was to investigate the influence of tobacco components tobacco extract TE nicotine and Cotinine on fungal colonization and biofilm formation on two acrylic denture resins Surface topography of the denture materials and alteration of fungal susceptibility to an anti fungal agent Nystatin of both planktonic and biofilm fungal cells grown on the acrylic resins were investigated Ivocap and Lucitone polished and roughened acrylic resin discs were fabricated and randomly assigned to control tobacco extract nicotine and Cotinine treated groups Candida albicans biofilm was prepared on the resin discs in the presence or absence of tobacco components The relative number of viable fungal cells was determined by the MTT assay Quantization of biofilm growth was performed by dry weight analysis Nystatin susceptibility minimum inhibitory concentration MIC was determined by microdilution method Statistical significance was evaluated using ANOVA followed by Fisher rsquo s test p lt The study showed that the roughened discs had significantly greater numbers of C albicansthan the polished discs Lucitone acrylic discs promoted significantly more fungal biofilm growth than Ivocap discs and the presence of tobacco components significantly enhanced biofilm formation on both types of acrylic resin Dry weight analysis showed similar results The Nystatin susceptibility assay showed that cells treated with tobacco components were more resistant to this antifungal agent