Aim: To investigate the effect of pro-inflammatory cytokines and black-pigmented Bacteroides on the expression of IL-6 gene in human pulp fibroblasts. Methodology: IL-1α tumour necrosis factor-α (TNF-α) and the supernatants of Porphyromonas endodontalis, P. gingivalis and Prevotella intermedia were used to evaluate IL-6 gene expression in human pulp fibroblasts. The levels of mRNAs were measured by the quantitative reverse-transcriptase polymerase chain reaction analysis. Results: Investigations of the time dependence of IL-6 mRNA expression in pro-infammatory cytokines-treated cells revealed a rapid accumulation of the transcript after 2 h of exposure and remained elevated throughout the 24-h incubation period. In addition, black-pigmented Bacteroides also induced IL-6 gene expression in human pulp fibroblasts. Conclusions: Pro-inflammatory cytokines and black-pigmented Bacteroides may be involved in developing pulpal inflammation through the stimulation of IL-6 production.