The Effect Of Electroactive P(VDF-TrFE) Membranes On Angiogenesis Of Human Umbilical Vein Endothelial Cells

BackgroundBone loss due to trauma,tumors,periodontitis,peri-implantitis,and other diseases remains a major challenge in implantology and periodontics.Currently,guided bone tissue regeneration is one of the most widely used bone augmentation protocols in clinical practice.The application of barrier membranes plays a critical role in the subsequent outcome of the procedure.Most barrier membranes are biologically inert materials that only provide a physical barrier and lack biological activity.Therefore,research to develop bioactive barrier membranes is of high clinical relevance.During bone tissue regeneration,neovascularization plays a significant role.Previous studies have shown bioelectrical signals play a facilitating role in the process of angiogenesis.However,the effects of materials with different electrical properties on angiogenesis are still unclear.Therefore,our group proposed preparing an electroactive material P(VDF-TrFE)and studying its effects on HUVECs’ activity and angiogenic function.The goal of this study was to lay the groundwork for the use of a new electroactive guided bone tissue regeneration membrane based on P(VDF-TrFE).ObjectiveThis project aimed to prepare electroactive P(VDF-TrFE)membranes with different surface potentials and investigate their effects on the activity,proliferation,migration,and angiogenic function of HUVECs in vitro.MethodsFirstly,P(VDF-TrFE)was prepared by the flow-delay method and polarized by oil bath polarization.The experimental groups were set up as negative poled group,positive poled group,nonpoled group,and blank control group.The physicochemical properties of the materials were examined using scanning electron microscopy and water contact angle.The crystalline shape of the materials was examined using Fourier transform infrared spectroscopy and X-ray diffractometry.Analysis of changes in electrical properties of materials by measuring surface potential.Cell activity was detected in direct contact experiments using live-dead staining,CCK-8 kit.Cell morphology was observed using fluorescence microscopy and migration of HUVECs was observed by scratching experiments.The effect of the materials on the tube formation ability of HUVECs was observed by tube formation assay.The effect of the materials on the expression level of angiogenesis-related factors of HUVECs was detected by qRT-PCR and western blot.Results1.After polarization,the P-phase content of P(VDF-TrFE)increased,and the surfacedisplayed different surface potentials.The polarization treatment did not affect the surface morphology and wettability of the materials(P>0.05).2.The results showed that the polarization treatment did not affect the cell activity,and the fluorescence staining of cell morphology showed cellular spreading on the surface of the materials.CCK-8 assay results showed that the proliferation rate of HUVECs was significantly promoted by P(VDF-TrFE)in the negative poled group as compared to that in the other three groups(P<0.05).The results of scratch experiments showed that the negative poled group significantly promoted cell migration(P<0.05).3.The results of the tube formation assay showed that the negative poled group significantly promoted the formation of tubular structures(P<0.05),and the qRT-PCR results showed that the expression levels of VEGF-A and CD31 genes significantly increased in the negative poled group(P<0.05).Similarly,the western blot results showed that the expression levels of the VEGF-A protein significantly increased in the negative poled group(P<0.05).Conclusions1.The surface of electroactive P(VDF-TrFE)membranes prepared in this experiment form different surface potentials and have good and stable electrical properties.2.The negative poled group of electroactive P(VDF-TrFE)membranes show good proangiogenic activity in vitro test.Clinical significanceElectroactive P(VDF-TrFE)membranes showed good cytocompatibility and proangiogenic activity in vitro tests.They are expected to be used as new electroactive barrier membranes for clinical use in the future.