Preparation Of IR780 And VCR-Loaded Plga Nanoparticles And Anti-nephroblastoma Cells Proliferation In Vitro

PARTⅠPREPARATION AND CHARACTERIZATIONS OF VCR LOADED PLGA NANOPARTICLESObjective: To prepare stable VCR-loaded PLGA nanoparticles(VPN)and measure its basic characteristics,and lay the foundation of the production of targeted nanoparticles.Methods: VPN were prepared by double emulsification method,and its general properties including morphology,particle size,structure were tested.The standard curve of VCR was drawn and the encapsulation rate and loading rate of VPN were calculated.The release of VPN in vitro was studied by the dialysis bag.Results: VPN showed milky white suspension when were dissolved in double steamed water.Under the optical microscope,VPN were spherical,uniform in size,regular in shape,and without aggregation and adhesion.The morphology of VPN were spherical or quasi-spherical with smooth surface,it had no obvious adhesion and the particle size distribution was uniform,the average particle size of VPN was(199.70±3.84)nm,the average zeta potential of VPN was(-1.93±0.17)m V,the average encapsulation efficiency of VPN was about 72.80%,and the average drug loading was 2.80%.It can be seen from the release experiment in vitro that the release behavior of VPN has two phases,sudden release and sustained release.Conclusion: The VCR-encapsulated VPN was successfully prepared which had a high drug encapsulation efficiency and prolonged the duration of drug action in this experiment.It laid a foundation for the preparation of targeted nanoparticles in the later stage.PARTⅡPREPARATION AND CHARACTERIZATIONS OF IR780 AND VCR-LOADED PLGA TARGETED NANOPARTICLESObjective: To prepare IR780 and VCR-loaded PLGA nanoparticles(IR780-VPN)and measure its general properties.Methods: The IR780-VPN was prepared by double emulsification method to detect their basic physical and chemical characteristics.The standard curve of VCR was drawn and the encapsulation rate and loading rate of IR780-VPN were calculated.The standard curve of IR780 was drawn and the specific targeting efficiency of IR780-VPN were calculated.The release of IR780-VPN in vitro was measured.Results: The average particle size of IR780-VPN was(290.82±3.22)nm,and the particle dispersion index(PDI)was 0.024,the average zeta potential of IR780-VPN was(1.16±0.87)m V.IR780-VPN showed stable physical and chemical properties,it was spherical,uniform in size,regular in shape,and without aggregation and adhesion under microscope.The morphology of IR780-VPN was spherical or quasi-spherical with smooth surface,and the particle size distribution was uniform.The average encapsulation efficiency of IR780-VCR-PLGA NPs was about 72.80%,the average loading efficiency was 2.80%,and the specific targeting efficiency was 91.30%.Conclusion: IR780-VPN showed stable physical and chemical properties with high encapsulation efficiency and loading efficiency on VCR.It also had a high specific targeting efficiency to IR780.PART Ⅲ STUDY ON SPECIFIC TARGETING TO NEPHROBLASTOMAAND ANTI-NEPHROBLASTOMA CELLS PROLIFERATION OF IR780-VPN IN VITROObjective: To verify specific targeting ability and anti-proliferation efficiency on SK-NEP1 cells of IR780-VPN in vitro.Methods: SK-NEP1 cells were used as tumor cell model to verify specific targeting ability and the anti-proliferation efficiency of IR780-VPN,SK-NEP1 cells were divided into 6 groups: group A:PBS(control group);group B: free VCR;group C: PLGA NPs;group D:VCR-PLGA NPs;group E: IR780-PLGA NPs;group F: IR780-VPN.Results: There was lots of IR780-VPN targeting to tumor cells from the trial of specific targeting to tumor cells.The cytotoxicity of group F was the highest under the same concentration(P<0.05),and there was no significant difference among the group A,group C and group E(P>0.05).Conclusion: IR780-VPN had a good ability of specific targeting to tumor cells and improved anti-proliferation efficiency in vitro.Lay the foundation for tumor molecular targeting research in vivo.