Construction And Characterization Of Pectin Doxorubicin Polymer Nanosystem Encased By Erythrocyte Membrane

The pectin-doxorubicin conjugates encapsulated by red blood cell membrane(PDC@RBC-NPs)was constructed,the PDC@RBC-NPs need to be characterized at first,and then the biocompatibility of which was studied.Finally,evaluating its cellular uptake and inhibition of hepatocarcinoma proliferation in vitro.The article is structured as follows:PDC-NPs are optimized nanoparticles based on the previous basis of the laboratory,which possess smaller size and are more conducive to internalized into the tumor site by EPR effect.We have prepared PDC@RBC-NPs by compressing PDC-NPs into erythrocyte vesicles using low-permeability extrusion.The average particle size and zeta potential of PDC-NPs and RBCm-vesicles were respectively measured by nanometer particle size analyzer,and visually observed by transmission electron microscopy(TEM).The results showed that PDC@RBC-NPs present concentric circles with an average particle size of 151.4 nm and a zeta potential of-16.0 mV.The stability test showed that PDC-NPswrapped with red blood cells are more stable than the one without,and the formed NPs could protect the drugs in the simulated human blood environment(pH 7.4).In vitro drug release experiments showed that PDC@RBC-NPs had a certain sustained release effect.Drugs get into the human body and enter the blood vessel primitively after intravenous injection,thus we evaluated the hemolysis rate of PDC@RBC-NPs via hemolysis rate measurement,and then detected the survival rate of HUVEC cells by MTT assay to evaluate the effect of PDC@RBC-NPs on vascular endothelial cells.Since doxorubicin has the serious toxic side effects including myocardial toxicity,MTT assay was carried out to evaluate the survival rate of PDC@RBC-NPs on human normal liver cell L02 and rat cardio myocyte H9c2 cells.The above experiments demonstrate that PDC@RBC-NPs retain a sustained release profile and have a better biocompatibility than free DOX and PDC-NPs.The drug,as an exogenous macromolecule,is easy for the body to trigger immune response and to be swallowed.As a result,the efficacy will be greatly reduced.In order to investigate the uptake of PDC@RBC-NPs by the immune system,we utilized PMA-induced THP-1-derived macrophages as a cell model to measure intracellular DOX fluorescence intensity through flow cytometry and observe the uptake of macrophages through fluorescence imaging.The results above showed that PDC@RBC-NPs wrapped in erythrocyte membrane caneffectively reduce the immunogenicity of PDC-NPs,improve the long-term circulation of drugs in vivo,and avoid the premature capture and clearance of PDC-NPs by immune cells,which is beneficial to protect the drug from transport in the body.Based on the previous research basis,we came to know that PDC-NPs possess a certain therapeutic effect on liver cancer so that the tumor proliferation was inhibited.Thee cell lines of liver cancer,HepG2,SMMC-7721,and BEL-7402 were selected for evaluation.Combining the MTT results of these cell lines,PDC-NPs after erythrocyte-encapsulated showed similar inhibitory effects on these ells with PDC-NPs,which indicate that PDC@RBC-NPs can decrease the immunogenicity of PDC-NPs while without affecting its normal inhibitory effect on hepatoma cells at the cellular level.