Effect Of Gold Nanoparticles On Morphology And Function Of HUVECs In Vitro Angiogenesis

Objective To observe the effects of gold nanoparticles (AuNP) on proliferation,migration and tube formation of human umbilical vein endothelial cells (HUVECs) invitro angiogenesis, as well as the changes in cell morphology during the process, andstudy the mechanism of AuNP on the inhibitory of angiogenesis.Methods AuNP and VEGF165were incubated overnight at4℃. The proliferationrate of HUVECs cell was detected by MTT. The ability of cell migration was assessedby using the modified of wound healing assay and transwell chamber assay. Then theability of tube formation was examined by capillary-like tube formation assay andchick chorioallantoic membrane (CAM). Moreover, we detected the morphology andultratructure of cell surface by atomic force microscope (AFM), observed the cellpseudopodia by scanning electron microscopy (SEM), and examined the changes ofactin filaments in the cytoskeleton by laser scanning confocal microscope (LSCM).Further, the phosphorylation of Akt in HUVECs on the VEGF165/VEGFR-2downstream was determined by Western Blot analysis.Results AuNP inhibited VEGF165-induced proliferation of HUVECs significantly,while AuNP alone did not. It inhibited VEGF165-induced migration and tubeformation of HUVECs, as well as neovascularization in vivo of CAM mode. AFMimage shown HUVECs were more active after treated with VEGF165, its appearedobvious pseudopodia, blurred edge with larger particles and rough surface in its cellmembrane, while treated with AuNP cells appeaered less active. After treated withAuNP, the pseudopodia and particles in cell surface of HUVECs were reduceddetected by SEM, the F-actin in HUVECs become disordered and chaotic thatexamined by LSCM. Further more, AuNP reduced the phosphorylation of Akt insignaling pathway of PI3K-Akt.Conclusion1.AuNP can inhibit VEGF165-induced proliferation, migration andtube formation of HUVECs, as well as neovascularization in vivo of CAM mode.2.AuNP can change the ultrastructure of cell surface, arrange the actin filamentsdisorderly, rearrange the cytoskeleton, and reduce pseudopodia of HUVECs indued by VEGF165.3. AuNP inhibit the angiogenesis possible via PI3K-Akt signalingpathway.