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Histological investigations have demonstrated that root canal sealers can induce mild to severe inflammatory alternations. However, there is little information on the precise mechanisms about root canal sealers-induced inflammatory reaction. Dysregulated cytokine productions at local disease sites have been considered to be major contributors to the development of inflammatory diseases. Interleukin (IL)-6 and IL-8 released have been reported to play an important role in the pathogenesis of inflammation. The aim of this study was to investigate the effects of root canal sealers N2 (zinc-oxide eugenol based) and AH Plus (epoxy resin based) on the expression of IL-6 and IL-8 mRNA gene in human osteoblastic cell line U2OS cells. The levels of mRNAs were measured by the semi-quantitative reverse-transcriptase polymerase chain reaction analysis. The exposure of quiescent U2OS cells to N2 and AH Plus resulted in the induction of IL-6 and IL-8 mRNA gene expression (p < 0.05). The intensity of IL-8 mRNA gene was found to be significant higher than IL-6 mRNA gene (p < 0.05). Taken together, the activation of IL-6 and IL-8 mRNA gene expression may be one of the pathogenesis of zinc oxide-eugenol based and epoxy resin based root canal sealers-induced periapical inflammation.