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. 2011 Aug 15;6(8):e23375. doi: 10.1371/journal.pone.0023375

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Miron et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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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).