Studies On The Preparation,drug Loading Properties And Cytotoxicity Of Protein/polyamidoamine Nanoparticles

Nanomaterial drug carrier,as an emerging carrier,has received more and more attentions.It is significant to develop nanocarrier system with safety and high performances for disease(especially for cancer)therapy.In this paper,combining the advantages of polyamidoamine(PAMAM)and human serum albumin nanoparticles(HSA NPs),the PAMAM,HSA NPs and HSA-PAMAM NPs were synthesized.The Doxorubicin(DOX)and Senkyunolide I(SEN I)were loaded by non-covalent interaction.The micromorphologies,surface charge and drug loading property of the prepared materials were investigated by various analytical methods.The cytotoxicity of the nanomaterials was investigated in detail using the HBMEC cells as a model.The in vitro blood-brain barrier(BBB)cell model was established to evaluate the performance of the HSA-PAMAM nanocomposites crossing the BBB.The main works are summarized as follows:1.The PAMAM was synthesized by divergence method.The DOX or SEN I was loaded into PAMAM by non-covalent interaction.The micromorphologies,surface functional groups,surface potential and particle size distribution of the prepared PAMAM were characterized by FITR,1H-NMR,gas chromatography,UV-vis spectroscopy,TEM and DLS.The efficiency of drug loading and entrapment into PAMAM were characterized by UV-vis spectroscopy.The results suggested that the loading amounts of DOX and SEN I on the PAMAM was 142.6?g mg-1 and 97.78?g mg-1.The entrapment efficiency of DOX and SEN I was 7.13%and 4.89%,respectively.The effect of PAMAM,PAMAM-DOX and PAMAM-SEN I hybrids on the cell viability of HBMEC cells was evaluated by MTT and WST-8 assays.The results show that the cell viability of HBMEC cells retains above 80%when the concentration is 100 ?g mL-1,indicating these materials have good biocompatibility to HBMEC cells.2.The HSA NPs with different size were prepared by desolvation method.The DOX or SEN I was loaded into HSA NPs by non-covalent interaction.The spectrum properties,micromorphologies,surface potential and particle size distribution of HSA NPs were characterized by UV-vis spectroscopy,SEM and DLS.The efficiency of drug loading and entrapment into HSA NPs were characterized by UV-vis spectroscopy.The results suggest that the loading amounts of DOX and SEN I in the HSA NPs was 45.17?g mg-1 and 17.52?g mg-1.The entrapment efficacy of DOX and SEN I was 75.29%and 87.60%,respectively.The yield of HSA-DOX NPs and HSA-SEN I NPs was 31.33%and 43.34%,respectively.The effect of the nanoparticles on the viability of HBMEC cells was evaluated by MTT and WST-8 assays.The results show that HSA NPs and HSA-SEN I NPs had good biocompatibility to HBMEC cells when the concentration is 100?g mL-1.However,HSA-DOX NPs exhibit dose-dependent cytotoxicity to HBMEC cells.3.The HSA-PAMAM NPs were prepared by desolvation method and DOX and SEN I was loaded into HSA-PAMAM NPs by non-covalent interaction.The micromorphology,surface potential and particle size distribution of nanoparticles were characterized by SEM and DLS.The efficiency of drug loading and entrapment into HSA-PAMAM was characterized by UV-vis spectroscopy.The results show that the loading amounts of DOX and SEN I on the HSA-PAMAM were 151.20?g mg-1 and 17.11?g mg-1,respectively.The entrapment efficacy of DOX,SEN I and PAMAM-DOX was 7.56%,88.16%and 54.93%,respectively.The yield of HSA-PAMAM-DOX NPs and HSA-PAMAM-DOX-SEN I NPs was 68.54%and 46.50%,respectively.The effects of these nanoparticles on the viability of HBMEC cells were evaluated by MTT and WST-8 assays.The results show that HSA-PAMAM NPs,HSA-PAMAM-DOX NPs and HSA-PAMAM-DOX-SEN I NPs had good biocompatibility to HBMEC cells when the concentration is 100?g mL-1.The in vitro BBB cell model was established using HBMEC cells and characterized by liquid interview leak test and HRP experiments.The permeability of HSA-PAMAM nanocomposites in BBB was evaluated by fluorescence spectroscopy.The result show that HSA-PAMAM nanocomposites labeled with FITC could cross the BBB,the permeability of HSA-PAMAM-DOX-FITC NPs and HSA-PAMAM-DOX-SEN I-FITC NPs in 24 h was 32.74%and 57.66%,respectively.It suggests that introducing of SEN I could significantly enhance the BBB permeability.