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Fig. 2 | BMC Musculoskeletal Disorders

Fig. 2

From: Sequential activation of the AKT pathway in human osteoblasts treated with Oscarvit: a bioactive product with positive effect both on skeletal pain and mineralization in osteoblasts

Fig. 2

Analysis of osteoblast mineralization. a Agarose gel electropherograms showing the RT-PCR banding patterns of DMP1-, BGLAP-, SPP1- and IBSP-mRNA in osteoclasts growing in MEM without OSC (lanes 1–4) and with 20 μg/ml OSC (lanes 5–12) over a cultivation period of 15 days. Expression of the housekeeping gene GAPDH was used as a loading control. (a1-a4) Densitographic evaluations of the electropherograms showed in 2a. Relative expression values of DMP1- (a1), SPP1- (a2), BGLAP- (a3), and IBSP-mRNA (a4), normalized by the amounts of GAPDH-mRNA expression were measured by Gel Doc gel documentation software. The cells were cultured in the absence (−, open bar) or the presence (+, closed bar) of OSC-containing medium. Colums represent mean values of four donors (−) and eight donors (+). b Mineralization of human osteoblasts from a primary culture. Scanning electron micrographs showing irregular- and rough cell surface after 15 days of cultivation in OSC-supplemented medium (b1) in contrast to osteoblasts growing in medium without OSC-supplementation (b2). Phase-contrast microscopic analysis showing calcium deposits (mineralization) in osteoblasts cultivated in OSC supplemented medium over cultivation periods of 5 days (b3, b4), 10 days (b5, b6), and 15 days (b7, b8) respectively. Alizarin-stained cells (b3, b5, b7); not Alizarin-stained cells (b4, b6, b8)

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