Melatonin Rescued Reactive Oxygen Species-Impaired Osteogenesis of Human Bone Marrow Mesenchymal Stem Cells in the Presence of Tumor Necrosis Factor-Alpha

Figure 2

Melatonin protected the osteogenic differentiation of BMSCs in the presence of TNF-α. BMSCs were cultured and underwent osteogenesis in osteogenesis medium containing vehicle (0.05 mol/L ethanol), TNF-α (10 ng/mL), or both TNF-α (10 ng/mL) and melatonin (100 μmol/L). (a) Mineralization of BMSCs after 14 days of osteogenesis was detected by ARS and photographed by microcopy in a representative sample. Scale bars: 200 μm. ARS staining was quantified as the absorbance at 405 nm. (b, c) The expression of runt-related transcription factor-2 (RUNX2) and osteopontin (OPN) was examined using (b) reverse transcription quantitative polymerase chain reaction and (c) Western blot after 3, 7, and 14 days of osteogenic differentiation. (d) The relative protein expressions of RUNX2 and OPN were established. The results are representative of three independent experiments. versus the vehicle group, versus the TNF-α group.

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