The increased release of prostaglandins (PG) within pulpal tissues is considered to play a pathogenic role during pulpal disease progression. The rate-limiting step in the formation of PG from arachidonic acid is catalyzed by cyclooxygenase (COX). COX-2 is an inducible enzyme believed to be responsible for PG synthesis at site of inflammation. The effect of proinflammatory cytokines on human pulp cells with special reference to COX-2 expression has not been reported earlier. The aim of the present study was to investigate the effects of interleukin (IL)-1alpha and tumor necrosis factor-alpha (TNF-alpha) on the expression of COX-2 mRNA gene and protein in cultured human pulp cells. Investigations of the time dependence of COX-2 mRNA expression in proinflammatory cytokines-treated human pulp cells revealed a rapid accumulation of the transcript, a significant signal first detectable 1 h after exposure. In addition, both IL-1alpha and TNF-alpha up-regulated COX-2 protein expression by human pulp cells. The kinetics of this response showed that COX-2 was detectable in cell lysates as early as 2 h post proinflammatory cytokines challenge and remained elevated throughout the 24-h incubation period. This suggests that one of the pathogenic mechanisms of pulpal inflammation in vivo may be the synthesis of COX-2 by resident cells in response to a proinflammatory cytokines challenge. COX-2 may play an important role in the regulation of prostanoid formation in the pathogenesis of pulpal inflammation. Taken together, we propose that the use of selective COX-2 inhibitors might provide a valuable tool in the control of pulpal inflammation.