Conditioned medium from human bone marrow stromal cells attenuates initial inflammatory reactions in dental pulp tissue

Abstract

Aim: To evaluate the effect of MSC-conditioned medium (CM) on the secretion of pro- and anti-inflammatory cytokines from dental pulp cells (hDPC) in vitro, and on the gene expression in vivo after replantation of rat molars. Materials and methods: hDPC were cultured in CM for 24 h, and the concentration of interleukin IL-10, IL-4, IL-6, and IL-8, regulated on activation, normal T Cell expressed and secreted (RANTES), and prostaglandin E2 (PGE2) in the media were measured by multiplex assay and ELISA, respectively. Expression of cyclooxygenase-2 (COX-2) was also examined by Western blot analysis after 24 h. Left and right maxillary first rat molars (n = 20) were elevated for 2 min and then replanted with or without application of CM into the tooth sockets. Levels of IL-1β, IL-10, IL-4, IL-6, and IL-8, and tumor necrosis factor-alpha (TNF-α) mRNA were evaluated by real-time qRT-PCR 3 and 14 days following tooth replantation. Results: The production of IL-8, IL-10, and IL-6, RANTES and PGE2 by cells cultured in CM was significantly higher than production by cells cultured in standard medium (DMEM). At day 3 following replantation in vivo, the levels of IL-1β and IL-6, and TNF-α mRNA were significantly lower in the CM-treated replanted teeth compared with control teeth. Further, at day 3, the IL-6/IL-10 ratio was significantly lower in the CM-treated replanted teeth compared with control. At day 14 following replantation, no differences in the mRNA ratios were detected between the pulp tissues of replanted and control teeth. Conclusions: These findings indicated that CM promotes secretion of pro- and anti-inflammatory cytokines from hDPCin vitro and attenuates the initial inflammatory response in the rat dental pulp in vivo following tooth replantation.