Figure 3.

Maintenance of osteogenic phenotype of pre‐osteoblasts through culture time in the presence of different osteogenic medium conditions. (A) Gene expression of osteogenic‐related markers, (i) bone sialoprotein 1/osteopontin (SPP1), (ii) alkaline phosphatase (ALPL), (iii) runt‐related transcription factor 2 (RUNX2), (iv) collagen type I (COL1A1) and (v) collagen type III (COL3A1) evaluated by RT‐PCR. Expression of target genes was normalized against GAPDH housekeeping gene and gene expression in all conditions was normalized to the basal condition (100 BM: 0 OM) of the correspondent day. Results are presented as mean ± SEM and statistically significant differences are shown as *, P < .04; **, P < .004; ***, P < .0002; δ statistically significant differences in comparison to the basal condition of the correspondent day. (B) Fluorescence microscopy images of osteopontin (scale bars, 50 μm). (C) Fluorescence intensity quantification of osteopontin at days 7 and 14