Study On The Preparation,Characterization,and Cell Absorption Of Genistein Albumin Nanoparticles

Genistein is a phytoestrogen,with a wide range of physiological activity and pharmacological effects,such as: anti-oxidation,anti-tumor,estrogen-like effect,protect the cardiovascular system,improve osteoporosis.However,Genistein is hardly soluble in water and has low bioavailability in vivo,which greatly influences the bioactivity of Genistein in vivo.In this paper,the Gen-BSA-NP were prepared by desolvation-chemical cross-linking method.The encapsulation efficiency and the drug-loading efficiency were used as the main evaluation indexes.The orthogonal experiment was used to optimize the preparation process.The optimum preparation conditions were as follows: the quality of BSA was 50 mg,the quality of Genistein was 6 mg,the volume of absolute ethanol was 20 m L,the volume of 0.25% glutaraldehyde was 150 μL,and curing at room temperature for 12 h.The entrapment efficiency of Gen-BSA-NP was(70.10±1.13)% and the drug-loading efficiency was(11.36±0.17)%,Transmission electron microscopy(Transmission Electron Microscope,TEM)and Zeta potential was used to characterize the structure and morphological analysis of Gen-BSA-NP.The results showed that the Gen could be encapsulated by BSA to form a regular spherical sphere and with a better dispersion in water,Z-Average(75.17±3.13)nm,Multi exponential dispersion coefficient(polydispersity index,PDI)(0.201±0.021),Zeta Potential(-35.43±0.29)mV.The Z-Average,PDI,Zeta Potential and TEM showed that the prepared nanosuspensions had good stability.The structure of Gen-BSA-NP was characterized by Fourier transform infrared spectroscopy(FTIR),X-ray diffraction(XRD)and differential scanning calorimetry(DSC).It is concluded that there are Genistein embedded in Gen-BSA-NP and the Genistein is present in an amorphous form.Stability and in vitro release of Gen-BSA-NP were analyzed.The results showed that the Gen-BSA-NP were stable at 37℃ and below,unstable high temperature(100 DEG C)and acidic conditions,The stability was the most stable at pH 7.4.Freeze-drying had no effect on the stability of the Gen-BSA-NP.Storage experiments showed that the Gen-BSA-NP were the most stable at 4℃.The in vitro release assay showed that the Gen-BSA-NP were slowly released in the simulated intestinal fluid and had a good sustained-release effect.The results of Caco-2 cytotoxicity test showed that BSA had no effect on the cell viability.With the further increase of concentration,the inhibitory effect of Gen-BSA-NP on Caco-2 cells was concentration-dependent,but with the concentration of Gen-BSA-NP reaches a certain level,the inhibitory ability of Gen-BSA-NP to cell growth was markedly superior in Gen monomer.The results of Caco-2 cell uptake showed that both Gen monomer and Gen-BSA-NP could be taken by Caco-2 cells,and Gen-BSA-NP was more easily absorbed by Caco-2 cells than Gen monomer.The results of Caco-2 cell absorption showed that Gen in Gen-BSA-NP had a higher rate of change than Gen monomer,and the time-dependence was stronger.The difference of Papp on both sides of the sample showed that the metabolism and transformation occurred in the process of transmembrane transport of Genistein.Gen-BSA-NP was more easily absorbed by Caco-2 cells than Gen monomer.