Stimulation of cytokines in osteoblasts cultured on enamel matrix derivative

Objective. The purpose of this study was to evaluate the influence of enamel matrix derivative (EMD) on the release of transforming growth factor beta 1 (TGF-beta 1), interleukin-6 (IL-6), insulin-like growth factor I (IGF-I), bone morphogenetic protein 2 (BMP-2), and osteoprotegerin (OPG) in human and mouse osteoblasts. Study design. Human MG-63 and mouse MC3T3-E1 cells were seeded onto 6-well culture plates at an initial density of 5,000/cm(2) and grown in Dulbecco's eagle medium (DMEM) with 10% fetal bovine serum for 24 h. Then cells were cultured either with 100 mu g/ mL EMD added to DMEM or with DMEM only. After 2, 5, and 9 days' incubation the culture medium was collected and analyzed by enzyme-linked immunosorbent analysis. Data were analyzed using Student t test. Results. The EMD treatment significantly increased the production of IL-6 and TGF-beta 1 (P < .05) at all time points. The release of OPG was also increased in mouse osteoblasts (P < .05). IGF-I and BMP-2 were not detected in both control and EMD-treated groups. Conclusion. This study suggests that the stimulatory effects of EMD on tissue regeneration are mediated by the upregulation of local mediators released by osteoblasts.