Study On Multiple Modification Graphene Oxide Delivery System Based On Transferrin And Folic Acid

Objective:The nano-carriers with targeting and good biocompatibility were prepared by using Transferrin?Tf?,Folic acid?FA?and Pluronic F68?PF68?modified Graphene oxide?GO?.Loading doxorubicin and determining drug-loading performance and preliminary anti-tumor activity.Method:1.Graphene oxide was prepared by modified Hummer's method.GO was modified with Transferrin and Folic acid to enhance tumor targeting of drug delivery system.GO was modified with Pluronic F68 to enhance biocompatibility.GO,GO-PF68,FA-GO-PF68,Tf-GO-PF68and Tf-FA-GO-PF68 was characterized by X-ray diffraction?XRD?,scanning electron microscope?SEM?,transmission electron microscope?TEM?,X-ray photoelectron spectroscopy?XPS?,Fourier transform infrared spectroscopy?FTIR?and ultraviolet-visible spectroscopy?UV-vis?.2.Doxorubicin was loading onto the nanocarrier and analyzing its drug-loading and drug-releasing properties.3.Human liver stellate cell line?7702?,human breast cancer cell line?MCF-7?,human breast cancer cell line?MDA-231?,hepatoblastoma cell line?HepG2?,human hepatoma cell line?SMMC-7721?and Macrophages?RAW264.7?were used as cell models.Cell lines toxicity of delivery systems and drug delivery systems were measured by MTT assay.4.Evaluation of carrier stability by measuring the dispersibility of the carrier in distilled water and DMEM.5.The anti-tumor activity of each drug-loading system was determined by MTT method.Results:1.GO,FA-GO-PF68,Tf-GO-PF68,Tf-FA-GO-PF68 were successfully prepared and characterized by XRD,SEM,TEM,XRD,FT-IR and UV-vis;2.The saturation loading of FA-GO-PF68 on doxorubicin was 72.5%,the cumulative release rate of DOX from FA-GO-PF68*DOX in the environment on pH=5.50 was 89.72%which was higher than the cumulative release rate in the environment on pH=7.40?43.2%?.The saturation loading of DOX from Tf-GO-PF68 on doxorubicin was 87.5%,the cumulative release rate ofDOX from Tf-GO-PF68*DOX in the environment on pH=5.50 was 54.02%which was higher than the cumulative release rate in the environment on pH=7.40?44.6%?;the saturation loading of Tf-FA-GO-PF68 on doxorubicin was 49.8%,and the cumulative release rate ofDOX from Tf-FA-GO-PF68*DOX in the environment on pH=5.50 was 71.4%which was higher than the cumulative release rate in the environment on pH=7.40?58.8%?.3.The cell toxicity assay showed that the survival rates of FA-GO-PF68,Tf-GO-PF68 and Tf-FA-GO-PF68 after treatment with 7702 cells for 24 h were91.16%,77.61%and 77.82%,the survival rates for 48 h was 92.25%,81.94%,80.44%.The cell toxicity assay showed that the survival rates of the carriers FA-GO-PF68,Tf-GO-PF68,Tf-FA-GO-PF68 acted on MCF-7 cells for 24 h were 79.37%,81.18%,100.54%,the survival rates for 48 h were 106.95%,100.67%,101.79%.The cell toxicity assay showed that the survival rates of the carriers FA-GO-PF68,Tf-GO-PF68 and Tf-FA-GO-PF68 on MDA-231 cells for24 h were 85.20%,77.19%,80.44%,the survival rates for 48 h were 75.66%,82.57%,91.22%.The cell toxicity assay showed that the survival rates of the carriers FA-GO-PF68,Tf-GO-PF68 and Tf-FA-GO-PF68 on HepG2 cells for 24h were 90.29%,79.99%,81.42%,and the survival rates for 48 h was 86.74%,85.85%,89.25%.The cell toxicity assay showed that the survival rates of the carriers FA-GO-PF68,Tf-GO-PF68 and Tf-FA-GO-PF68 on SMMC-7721 cells for 24 h were 109.46%,75.36%,84.11%,the survival rates for 48 h were84.81%,79.18%,85.18%.The cell toxicity assay showed that the survival rates of the carriers FA-GO-PF68,Tf-GO-PF68,and Tf-FA-GO-PF68 for 24 h on RAW264.7 cells were 80.95%,75.59%,the survival rates for 48 h were 80.02%,80.68%,80.02%,82.65%;4.FA-GO-PF68,Tf-GO-PF68,Tf-FA-GO-PF68 have good stability in water and DMEM.;5.Drug-loading systems FA-GO-PF68*DOX,Tf-GO-PF68*DOX and Tf-FA-GO-PF68*DOX acted on SMMC-7721 cells for 24 h and the IC₅₀0 values were 8.692,38.813 and 59.95?g/mL.The IC₅₀0 values of SMMC-7721 cells for48 h were 2.057,0.553 and 0.875?g/mL.The drug system FA-GO-PF68*DOX,Tf-GO-PF68*DOX and Tf-FA-GO-PF68*DOX acted on HepG2 cells for 24 h and the IC₅₀0 values were 20.698,10.881 and 3.278?g/mL.The IC₅₀0 values of HepG2 cells for 48 h were 13.242,3.314 and 5.957?g/mL.Conclusion:1.Successfully prepared graphene oxide by improved Hummers method;2.GO,GO-PF68,FA-GO-PF68,Tf-GO-PF68 and Tf-FA-GO-PF68 were characterized by XRD,SEM,TEM,XPS,FT-IR and UV-vis which showed that PF68,FA and Tf were successfully modified with graphene oxide and enhance GO efficiency and specificity;3.The drug-loading and drug release performance assay results of FA-GO-PF68*DOX,Tf-GO-PF68*DOX and Tf-FA-GO-PF68*DOX showed that FA-GO-PF68,Tf-GO-PF68 and Tf-FA-GO-PF68 carriers all had good drug-loading and drug-releasing properties.It's said that FA-GO-PF68,Tf-GO-PF68 and Tf-FA-GO-PF68 all can be used as drug carrier for DOX;4.Cytotoxicity test results show that FA-GO-PF68,Tf-GO-PF68,Tf-FA-GO-PF68 pair 7702,MCF-7,MDA-231,HepG2,SMMC-7721 and RAW264.7 has no obvious cytotoxicity;5.FA-GO-PF68,Tf-GO-PF68,Tf-FA-GO-PF68 have good stability in water and DMEM.6.FA-GO-PF68*DOX,Tf-GO-PF68*DOX,Tf-FA-GO-PF68*DOX toxicity test on tumor cells indicates that all three drug-loading systems have certain anti-tumor activity.