Figure 3.

Osteoclast viability, differentiation and transcription in culture after exposure to serum harvested from mice either raised under control conditions or treated with pomegranate peel extract. * p < 0.05 PGPE versus Control condition. (A) Cell proliferation test performed on RAW264.7 cell line cultured in minimal medium (2% Fetal Bovine Serum, FBS), supplemented with serum from mice exposed to either physiological serum (1.25% FBS + 0.75% Control mice serum) or pomegranate peel extract, PGPE (1.25% FBS + 0.75 PGPE mice serum) after 1 and 2 days of treatment. Results are expressed as mean ± standard error of the mean (SEM) (n = 6). (B) Tartrate resistant acid phosphatase, TRAP activity of RAW264.7 cells cultured in optimized medium with 10% FBS or with 7.5% FBS + 2.5% mice serum from animals fed either physiological serum (Control) or pomegranate peel extract (PGPE), after 3 days of treatment. Results are expressed as mean ± SEM (n = 8). (C) Transcriptomic analysis of the main osteoclast markers in RAW264.7 cells cultured in optimized medium after 2 days of treatment with serum from mice exposed to either physiological serum (Control) or pomegranate peel extract (PGPE). Transcriptomic analysis of the Raw264.7 mRNA levels determined by Taqman Low density Arrays are presented as fold change compared to Control condition (fold change = 1) as mean ± standard deviation (SD) (n = 6). Gene: CaM: calmodulin; CCR2: chemokine receptor 2; CTR: calcitonin receptor; CTSK: cathepsin K; FOS; IFN β1: interferon β1; MMP9: metalloproteinase 9; Myc, RANK: receptor activator of nuclear factor-κB; Relb; c-SRC: and TRAF6.