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. 2013 Mar 5;8(3):e58580. doi: 10.1371/journal.pone.0058580

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© 2013 Onami 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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The activity of vasohibin-1 was confirmed by an in vitro endothelial cell tube formation assay. Vasohibin-1 suppressed the HUVEC tube formation in a dose-dependent manner. Representative results of HUVEC tube formation treated with 2 nM VEGF combined with 0 (A), 0.2 (B), 2 (C), and 10 nM vasohibin-1 (D) are shown. Bars indicate 100 μm. The released vasohibin-1 from the device showed comparable results to native activity (E). Significant suppression of HUVEC tube formation was observed in released vasohibin-1 when compared to those treated with NVDD (F) and with only 2 nM VEGF without vasohibin-1 (G). (H) shows the average of each experiment; significantly fewer CD31-positive points were observed in released vasohibin-1-treated wells when compared to those of the vehicle released from NVDD (p<0.0001) or the VEGF-treated control (p<0.0001). The vasohibin-1 released from the device showed activity comparable to the native vasohibin-1. Vertical bar indicates total length of tube formation. NVDD: non-vasohibin-1 (vehicle) delivery device, 10VDD: 10 μM vasohibin-1 delivery device.