EMD increases alkaline phosphatase activity in osteoblast clusters.

<p>At time points 1, 3, 5, 7 and 10 days, osteoblasts were fixed and stained for alkaline phosphatase. A) EMD significantly increased alkaline phosphatase activity in clustered regions 5 days post seeding. After 10 days, complete staining was observed on samples coated with EMD when compared to control samples. (bar = 1000 µm) B) 10 fields of view per sample were captured and percentage area of staining calculated. Data represent means ± SE (results from 3 independent experiments). Significant increases in alkaline phosphatase activity were observed on EMD treated samples at 5, 7 and 10 days post seeding. C) mRNA was extracted and realtime PCR was performed using specific primers for alkaline phosphatase. Samples pre-coated with EMD showed significant mRNA levels 3, 5 and 7 days post seeding when compared to control groups (p<0.05). Data represent means ± SE (results from 3 independent experiments).