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Figure 2 | BMC Complementary and Alternative Medicine

Figure 2

From: Anti-angiogenic potential of an ethanol extract of Annona atemoya seeds in vitro and in vivo

Figure 2

EEAA inhibits angiogenesis in vitro and in vivo. (A and B) Cell proliferation and viability of HUVECs after 24 h of EEAA treatment. Cells were treated with 25–100 μ g/mL EEAA for 24 h. Viable and dead cells were discriminated and counted using an automated cell counter, as described the Methods section. (C) Representative images depicting HUVEC migration after EEAA treatment with the indicated concentrations. Wound closures were measured and recorded at 0 and 18 h after injury. (D) Representative images depicting the formation of capillary-like tube structures on Matrigel by HUVECs after 8 h of EEAA treatment with the indicated concentrations. The effects of EEAA on the formation of capillary-like tube structures by HUVECs such as the changes in tube length and the number of joints, were quantified as described in the Methods section. The data are presented as the relative means ± S.D. of at least three independent experiments compared with the vehicle *P < 0.05, **P < 0.01, and ***P < 0.001 compared with the vehicle-treated groups. (D and E) (E) Effect of EEAA on angiogenic factor-derived vessel formation in vivo. Angioreactors filled with BME premixed with combinations of the angiogenic factors VEGF/FGF2 and various concentrations of EEAA (0–100 μ g/mL) were implanted into the dorsal flanks of nude mice. After 12 days, the implanted cylinders were harvested (left panel), and vascular endothelial cells that had migrated into the BME gels of the bioreactors were quantified using FITC-lectin detection (right panel). The negative control represents the bioreactors without VEGF/FGF2 inducers or EEAA. Relative angiogenesis was normalized to the mean of the negative control. *P < 0.05, and ***P < 0.001 compared with the positive control group.

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