Effects Of Nano-silica On Proliferation,Migration And Formation Of Tubular Structure Of Lymphatic Endothelial Cells

Objective: To study the effects of silica nanoparticles(silica nanoparticles,SiNPs)on proliferation,migration and formation of tubular structures of human lymphatic endothelial cells(human lymphatic endothelial cells,HLECs),and to explore the effect and mechanism of SiNPs on lymphangiogenesis.Methods: Suspension of silica was prepared and its morphology was observed by transmission electron microscopy(TEM).Nanoparticle size analyzer measures the particle size of SiNPs in solution.HLECs were cultured and intervened with SiNPs with a particle size of 30 nm.The experiment was divided into four groups,control groups,10 ?g/m L SiNPs group,20 ?g/m L SiNPs group,and 40 ?g/m L SiNPs group.Cell proliferation assay was used to detect the effect of SiNPs on the proliferation of HLECs.The Transwell migration assay measures the effect of SiNPs on the migration of HLECs.Matrigel experiments were performed to examine the effect of SiNPs on the formation of tubular structures of HLECs.The expression of Vascular Endothelial Growth Factor C(VEGF-C),Vascular Endothelial Growth Factor D(VEGF-D)and Vascular Endothelial Growth Factor Receptor-3(VEGFR-3)m RNA was detected by RT-PCR.The expression of VEGF-C,VEGF-D and VEGFR-3 protein was detected by.Western-blot methodResults: SiNPs are irregularly spherical,and the particle size is mainly concentrated at about 30 nm.In the cell experiment,the proliferation of cells in the10?g/m L SiNPs group was not significantly different after treatment with SiNPs for24h(P>0.05).The proliferation of cells in the 20?g/m L SiNPs group and 40?g/m L SiNPs group was obvious.The decrease(P<0.05),the proliferation ability of HLECs in each intervention group decreased after 36 h and 48 h compared with the control group(P<0.05).Compared with the control group,the migration of HLECs in the10?g/m L SiNPs group and the 20?g/m L SiNPs group was significantly increased(P<0.05),while the number of migrated cells in the 40?g/m L SiNPs group was not significantly changed(P>0.05).Compared with the control group,the number of tubular structures in each intervention group increased significantly(P<0.05).There was no significant difference in the number of tubular structures between the intervention groups(P>0.05).The expression levels of VEGF-C,VEGF-D protein and m RNA in the 40?g/m L SiNPs group decreased with the increase of SiNPs concentration,which was significantly lower than that of the control group(P<0.05),while the 10?g/m L SiNPs group compared with the control group.There was no significant difference(P>0.05).The expression levels of VEGFR-3 protein andm RNA in the 10?g/ml SiNPs group were significantly higher than those in the control group(P<0.05).When the concentration of SiNPs was 40?g/m L,the expression levels of VEGFR-3 protein and m RNA were not significantly different from those of the control group(P>0.05),and the protein and m RNA expression levels were significantly lower than those of 10 and 20?g/ml SiNPs.(P<0.05).Conclusion: SiNPs can affect the proliferation,migration and tubular structure of HLECs by regulating the expression of related factors such as VEGF-C,VEGF-D and VEGFR-3.