| Objective Fibrosarcoma is a common malignant soft tissue tumor.t present,the treatment mode is mainly surgical resection,with chemotherapy as adjuvant treatment,but the chemotherapeutic drugs have poor oral absorption,low selectivity,and are prone to serious adverse reactions.In order to reduce toxic side effects,new drug carrier delivery system has become a research hotspot.Because polymer nanoparticles(NPs)can protect drugs from degradation,improve drug utilization and control drug release,they have been widely developed as drug carriers.Red blood cells(RBC)have also been used in drug carrier research because of their advantages of high bioavailability,good biocompatibility and long cycle life.Therefore,the purpose of this paper is to improve the therapeutic effect of chemotherapeutic drugs on tumor by combining the two drug loading methods.Methods The HPLC method of VCR was established according to Chinese Pharmacopoeia,and the method was verified.VCR DSPE-PEG2000-PLGA nanoparticles(VCR-NPs)were prepared by nanoprecipitation method.The particle size and entrapment efficiency were used as evaluation indexes,the appropriate factor level was selected by single factor investigation,and the VCR-NPs prescription was screened by Box-Behnken effect surface method to obtain the optimal prescription.The Zeta potential,appearance morphology,stability and in vitro release characteristics of the VCR-NPs were evaluated.VCR-NPs was incubated on the surface of red blood cells(RBC)by co-incubation method to prepare RBC-VCR-NPs.The appropriate incubation conditions were obtained by single factor test and drug loading as the evaluation standard.The proportion of prescription dosage of VCR-NPs and RBC was screened by shear force test,and the optimal RBC-VCR-NPS was determined by Flow Cyto Metry(FCM).The NGR-RBC-VCR-NPs was obtained by inserting the target NGR into the surface of RBC membrane by lipid insertion method to enhance targeting.The targeted effects of RBC-VCR-NPs and NGR-RBC-VCR-NPs on HT-1080 cells were observed by confocal laser scanning microscope.And the antiproliferation ability of VCR,RBC-VCR-NPs and NGR-RBC-VCR-NPs against HT-1080 cells in vitro was investigated by MTT method.Results For reference to the Chinese Pharmacopoeia 2015,the linear relation of VCR concentration in the range of 12.5~125 μg/m L is Y = 0.0194 X-0.1184,r2 = 0.9991.The RSD values of intraday and daytime precision of low,medium and high concentrations were less than 2%.RSD values of different concentrations were less than 2%.The recovery rate of VCR solution with low,medium and high concentration is between 98.06% and 100.83%,and RSD values of each concentration were less than 2%.The optimum formulation of VCR-NPs was as follows: acetone 2 m L,VCR 4 mg and DSPE-PEG2000-PLGA 40 mg.The morphology of NPs was spherical.The average particle size was 167.5 nm、zeta potential was-26.4 m V,the average encapsulation efficiency was 60.09% and the average drug loading was 3.21%.The stability of the VCR-NPs was good within 60 h by means of the stability instrument.VCR released 84.27% at 12 h and 75.29% at 120 h.The optimal incubation condition of RBC-VCR-NPs was 1 h at 37 ℃.Through the method of applying shear force in vitro,combined with the experimental results of phosphatidylserine exposure rate,the optimal prescription proportion was NPs:RBC=6:1 and the NGR-RBC-VCR-NPs average drug load was 0.531 mg/m L RBC.After treatment,the red blood cells were in good health,and the nanoparticles were successfully adsorbed on the surface of red blood cells.The targeting of NGR-RBC-VCR-NPs to cell HT-1080 was significantly higher than that of RBC-VCR-NPs.MTT results show that the anti-proliferative ability of NGR-RBC-VCR-NPs group acting on HT-1080 cells was significantly enhanced compared with the VCR group and the RBC-VCR-NPs group.And with the increase of drug concentration,the proliferation inhibition effect of NGR-RBC-VCR-NPs on tumor cells was enhanced.It is indicating that the targeting of the NGR-RBC-VCR-NPs to the tumor is significantly enhanced,thus improving the therapeutic effect of the drug.Conclusion VCR-NPs was prepared by nano precipitation method.The encapsulation efficiency and drug loading were stable,the particle size was small and the morphology was uniform.DSPE-PEG2000-PLGA nanoparticles were connected to the surface of red blood cells by phospholipid insertion.The red blood cells were in good condition without obvious damage.Improved the targeting of the carrier by connecting the NGR target.NGR-RBC-NPs can be targeted to HT-1080 cells.It had strong anti proliferation ability to HT-1080 cells and can improve the therapeutic effect of VCR. |
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