P178 - TNF-α and IL-1β primed Fetal cartilage Stem Cell-derived conditioned media alleviates inflammatory markers in SW982 cells
Abstract
Purpose
Many biologic effects of stem cells have been reported to be due to the secretome. In this study, we investigated whether the conditioned medium (CM) derived from fetal cartilage stem cells (FCSCs) priming reduced the inflammatory response of osteoarthritis (OA).
Methods and Materials
FCSCs priming was carried out by pro-inflammatory cytokine including Poly(I:C), IFN-γ, TNF-α and IL-1β. For synovial inflammatory model in vitro, SW982 were stimulated with TNF-α (25 ng/ml) and IL-1β (10 ng/ml). After 24 h incubation, inflammatory SW982 were treated with normal FCSCs-CM or primed FCSCs-CM for another 24 h. To determine the anti-inflammatory effect of FCSCs-CM, inflammatory gene (TNF-α, IL-1β and IL-6) expression of SW982 was examined by qRT-PCR. The level of PGE2 was measured by ELISA for evaluate the secretion of key inflammatory cytokine. To evaluate the priming effect of FCSCs-CM, anti-inflammatory related gene expression was measured with qRT-PCR, western blotting, protein array.
Results
After priming of FCSCs, expression of growth factors (IGF-1, TGF-β1 and FGF-2) and anti-inflammatory factors (IL-10, Leptin, IL-11, IDO-1, and IL1RA) was significantly higher than that of normal FCSCs. In the inflammatory SW982, primed FCSCs-CM showed higher anti-inflammatory ability than normal FCSCs-CM by inhibiting the expression of TNF-α, IL-1β and IL-6. The levels of PGE2 were significantly reduced in the treatment group with primed FCSCs-CM compared to normal FCSCs-CM. In addition, qRT-PCR results indicated that priming conditions of TNF-α (10 ng/ml) and IL-1β (15 ng/ml) effectively enhanced the anti-inflammatory ability of FCSCs-CM. In this study, in vivo immunomodulatory effects of TNF-α and IL-1β primed fetal cartilage stem cell-derived conditioned media alleviates osteoarthritis in rat model.
Conclusion
These studies suggest that TNF-α and IL-1β stimulate the fetal stem cells most effectively and secrete anti-inflammatory agents. TNF-α and IL-1β-primed FCSCs-CM show greater ability to inhibit inflammatory events in SW982 than normal by increasing secretion of anti-inflammatory cytokines.
