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. 2014 Sep 8;34(5):1201–1208. doi: 10.3892/ijmm.2014.1926

Figure 4.

Figure 4

Excess β-glycerophosphate causes non-specific mineralisation and decreases tissue non-specific alkaline phosphatase (TNAP) expression in mouse calvarial osteoblast cultures. All cells were cultured in αMEM with heat-inactivated FCS (HI FCS) and 50 μg/ml ascorbate. (A) Low power whole well scans and higher resolution phase contrast microscopy images showing alizarin red stained mouse osteoblast cell layers cultured in αMEM with 0–10 mM β-glycerophosphate. In the absence of β-glycerophosphate mineralisation does not occur, whilst 2–5 mM β-glycerophosphate results in the selective mineralisation of the collagenous matrix. The 10 mM β-glycerophosphate results in widespread, non-specific dystrophic mineralisation of cell layers. Scale bars: whole well, 5 mm; phase contrast micrographs, 500 μm. (B) The 10 mM β-glycerophosphate reduced TNAP expression by 40%. Values are the means ± SEM (n=6 replicate wells), *p<0.05.