Barley Seedling Extracts Inhibit RANKL-Induced Differentiation, Fusion, and Maturation of Osteoclasts in the Early-to-Late Stages of Osteoclastogenesis

Figure 5

BSE impairs the osteoclastic function of giant multinucleated cells. (a) The mRNA expression of cathepsin K was evaluated during osteoclast differentiation in the absence or presence of BSE (3 μg/ml) using real-time PCR. The relative fold change of the mRNA expression level is presented in comparison to the control (no RANKL-treated). HPRT was used as the internal control. (versus the vehicle control). (b) The mature osteoclasts were plated on bone biomimetic synthetic surface and treated for 24 hrs with BSE (3 μg/ml). The cells were fixed, permeabilised, and stained with TRAP. TRAP+ MNCs formation was visualised under a light microscope (top images). The resorption areas were removed from the cells and photographed under a light microscope (bottom images). (c-d) The form (as visualised in (b)) was counted as the number of TRAP+ MNCs (nuclei > 10; (c)), and the resorptive areas (%) were quantified using the ImageJ program (d). (versus the vehicle control). One representative result achieved from three independent experiments yielding similar results is shown.

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