The Construction Of Multifunctional Nanoparticles And Anti-Glioblastoma Research

Glioma is the most common malignant tumors of the central nervous system.Due to its malignant invasive growth,it was different to achieve the better therapeutic effect.Nowadays,photothermal therapy,photodynamic therapy,gene therapy,et.al,was gradually being applied to antiglioma.But none of the single treatment could completely satisfy the clinical treatment.Therefore,the multimodal therapy becomes the new strategy for glioma therapy.In this topic,two delivery systems were constructed in order to achieve better effect of glioma treatment.First,silica-coated gold nanorods were loaded with IR795 to form AuNRs@SiO2-IR795 nanoparticle as the passive targeting delivery systems combining photothermal and photodynamic therapy.Then,cationic liposomes carried gold nanorods,doxorubicin,YAP-siRNA and ANG2 to build a targeting drug delivery system mediated by low density lipoprotein receptor related protein receptor(LRP).It could achieve dual target to endothelial cells and glioma.The main content of the project includes four partsas follows:First,the synthesis and characterization of AuNRs@SiO2-IR795.The gold nanorods with different aspect ratio were prepared and the AuNRs@SiO2 was carried out with amino modified for the linking of IR795 dye.The effect of several factors on the fluorescence properties of IR795 was researched,such as the aspect ratio of gold nanorods,the molar ratio of gold nanorods/IR795 and the silica shell thickness.We found the best formula as AuNRs-3.3@SiO2(12.2 nm)-IR795,which has 51.7 folds fluorescence enhanced.Second,the antitumor efficiency of AuNRs@SiO2-IR795 nanoparticles in vitro.Fluorescent images,dark field microscope and ICP-MS were used to examine the uptake of nanoparticle in tumor cells.The ability of AuNRs@SiO2-IR795 to generate singlet oxygen and photothermal efficiency were detected by ABDA method and thermal imager.Then,MTT method was used to determine the cell survival rate incubating with a variety of nanoparticles under laser irradiation.According to the results,AuNRs@SiO2-IR795 had good biological compatibility and strong antitumor effect with PDT/PTT.Third,the synthesis of ANG2-Au-DOX-LP/siRNA targeting drug delivery system.Use the FT-IR and 1H-NMR for characterization of DSPE-PEG2000-ANG2.ANG2-Au-DOX-LP nanoparticle is determined by the single factor investigation to finding the best prescription,and we determined the size,zeta potential,encapsulation efficiency,drug loadings and release study in vitro.After incubation with siRNA,we get the ANG2-Au-DOX-LP/siRNA targeting drug delivery system and agarose gel electrophoresis showed that siRNA was well load on the nanoparticles.Fourth,Antiglioma efficiency of ANG2-Au-DOX-LP/siRNAi in vitro.Using laser confocal microscope and flow cytometry to examine the cell uptake.The Westen blot and QPCR was applied to test the gene silencing effect of YAP.MTT method were used to detect the effect of various drug delivery systems for cell proliferation.We established orthotopic glioma model.Results showed that ANG2-Au-DOX-LP/siRNA can simultaneously delivery DOX and siRNA into cells and the YAP mRNA and protein levelsin U87MG cells was significantly dropped.The combination therapy was better than single therapy.Moreover,ANG2-Au-DOX-LP/siRNA could effectively penetrate the blood-brain barrier and concentration in brain at 6 h.Preliminary antiglioma efficiency in vivo showed that ANG2-Au-DOX-LP/siRNA can significantly enhance the antiglioma efficency.