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. 2021 Jun 25;9:665886. doi: 10.3389/fcell.2021.665886

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Copyright © 2021 Janebodin, Chavanachat, Hays and Reyes Gil.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Alkaline phosphatase and Alizarin Red S staining in CopGFP control and VEGFR-2 shRNA DPSCs after a culture in osteo-odontogenic BMP-2 media. Alkaline phosphatase staining demonstrated a dissimilar result between CopGFP control (A) and VEGFR-2 shRNA DPSCs (B) after osteo-odontogenic induction in BMP-2 media. Abundant positive alkaline phosphatase stained cells (in dark-bluish purple) were shown in CopGFP control DPSCs, suggesting DPSCs undergoing osteo-odontogenesis (C,D). Few slightly positive stained for alkaline phosphatase staining were found in VEGFR-2 shRNA DPSCs (E,F). Alizarin Red S staining demonstrated a different result between CopGFP control and VEGFR-2 shRNA DPSCs after osteo-odontogenic induction in BMP-2 media. Several Alizarin Red S positive mineralized nodules were detected in differentiated CopGFP control DPSCs (G). There was absence of mineralized nodule found in differentiated VEGFR-2 shRNA DPSCs (H). A representative figure reveals a low magnification of Alizarin Red S positive mineralized nodules in the CopGFP control (I). The higher magnification of the nodule showed cuboidal-like cells, resembling osteo-odontoblast-like cells (J). VEGFR-2 shRNA DPSCs did not show any mineralized nodule formation (K) but they were undifferentiated spindle-shaped (L). Scale bars indicate 100 μm.