TGF-β1 stimulates cyclooxygenase-2 expression and PGE production of human dental pulp cells: Role of ALK5/Smad2 and MEK/ERK signal transduction pathways.

Background/purposes: TGF-β1 is an important growth factor that may influence the odontoblast differentiation and matrix deposition in the reactionary/reparative dentinogenesis to dental caries or other tooth injuries. TGF-β1 exerts its effects through various signaling pathways, such as Smads and MAPKs. Cyclooxygenase-2 (COX-2) is a membrane-associated enzyme that produces prostaglandin E (PGE) at sites of pulpal injury and inflammation, which leads to tissue swelling, redness and pain. The purposes of this study were to investigate the differential signal transduction pathways of TGF-β1 that mediate COX-2 stimulation and PGE production in dental pulp cells.

Methods: Pulp cells were exposed to TGF-β1 with/without SB431542 (an ALK5/Smad2 inhibitor) and U0126 (a MEK/ERK inhibitor). MTT assay was used to estimate cell viability. Enzyme-linked immunosorbent assay (ELISA) was used for measurement of PGE levels. RT-PCR and western blot were used to determined COX-2 mRNA and protein, respectively.

Results: Exposure to TGF-β1 (1-10 ng/ml) increased the COX-2 mRNA and protein level of cultured pulp cells. Exposure to TGF-β1 (0.1-10 ng/mL) significantly stimulated PGE production of dental pulp cells. Under the pretreatment of SB431542, the stimulatory effect of TGF-β1 on COX-2 level of pulp cells was inhibited. Similarly, U0126 also partly inhibited the TGF-β1-induced COX-2 expression.

Conclusion: TGF-β1 increased the COX-2 and PGE level of cultured pulp cells. The effect of TGF-β1 on COX-2 protein expression was associated with ALK5/Smad2/3 and MEK/ERK pathways. These events are important in the early inflammation, repair and regeneration of dental pulp in response to injury.