Applied Research Of Multifunctional Nanocarriers In Hepatocellular Carcinoma

BackgroundHepatocellular carcinoma(HCC)is the most common malignant tumor in the world.It is the sixth most common malignancy and the third most common cause of tumor death.Most HCC patients are already in the middle and advanced stages when they are diagnosed and cannot undergo radical treatments such as liver transplantation,surgical resection,and local percutaneous tumor ablation.At present,sorafenib(SRF),lenvatinib,donatinib,atilizumab plus bevacizumab are the first-line standard treatments for patients with advanced HCC,which could only prolong the survival time of patients for several months due to the serious decline of low tumor response rate,low bioavailability,and multiple toxicities.Nanoparticulate systems has great advantages in reducing the toxic and side effects of drugs and improving the treatment effect.At the same time,it also has many functions such as good biocompatibility,image tracing possibilities,and controlled release of carried drugs.This article provides a promising approach to realize supersensitive NIR-controlled drug release,photothermal therapy(PTT)and synergistic chemodynamic therapy(CDT)for cancer therapy.Ojective:A multifunctional nanocarrier SRF@CuFe2O4@PEA81 NPs was synthesized using mesoporous CuFe2O4 as the nucleo-loaded drug SRF and amphiphilic temperature sensitive polymer PEA81 as the outer layer.The drug delivery efficiency,distribution characteristics,anti-tumor effect and imaging ability of the nanoparticles for HCC were studied.Materials and Methods:1.Preparation and characterization of SRF@CuFe2O4@PEA81 NPs.Synthesize mesoporous CuFe2O4 and use it as a carrier to support SRF.Use X-ray diffraction(XRD)to verify the crystal phase and internal structure of mesoporous CuFe2O4.Use the Brunauer Emmett Teller(BET)and UV-vis spectrum to evaluate the ratio,drug loading(DL)and the entrapment efficiency(EE)of mesoporous CuFe2O4,test the photothermal effect of mesoporous CuFe2O4;prepare amphiphilic temperature sensitive polymer material PEA81,using 1H Nuclear magnetic resonanc(1H NMR)to analyze the chemical composition of the nanocarrier;using transmission electron microscope(TEM)to evaluate the shape,morphology and element distribution of SRF@CuFe2O4@PEA81 NPs.The drug release characteristics,PTT and CDT effects of SRF@CuFe2O4@PEA81 NPs were investigated by simulation experiments.MRI was used to evaluate the image tracer ability of SRF@CuFe2O4@PEA81 NPs.2.In vitro experiments.Cyanine-loaded nanocarrier Cy@CuF e2O4@PEA81 was co-located with lysosomal probe to investigate the uptake of Hepa1-6 cells.CDT reaction catalyzed by nanocarrier was evaluated with ·OH probe.CCK-8,(Calcein-AM/PI)and flow cytometry were used to analyze the cytotoxicity and antitumor effect of SRF@CuFe2O4@PEA81.3.In vivo experiments.C57BL/6 mice subcutaneous tumor model was established via subcutaneous injection of Hepa1-6 cells.In vivo fluorescence imaging and MRI imaging were used to analyze the distribution of SRF@CuFe2O4@PEA81 NPs in mice and to evaluate the passive targeting of NPs to tumors.The efficacy of SRF@CuFe2O4@PEA81 NPs in the treatment of HCC was evaluated by tumor growth inhibition assay,histopathological examination,TUNEL staining,and photothermal imaging.Results:1.SRF@CuFe2O4@PEA81 was successfully synthesized;TEM results showed that SRF@CuFe2O4@PEA81 NPs has a uniform particle size with an average of 110 nm and a near-spherical structure.SRF@CuFe2O4@PEA81 nanocarrier loaded with SRF was 14.49%and the entrapment efficiency was 72.45%.T2 relaxivity of SRF@CuFe2O4@PEA81 was 8.933 mL/(μg·s).Under the irradiation of 808nm near-infrared laser,CuFe2O4 can successfully generate photothermal effect.On the one hand,photothermal effect can accelerate the decomposition of H2O2 to generate OH.On the other hand,the resulting photothermal effect changes the structure of the amphiphilic temperature sensitive polymer material PEA81 and successfully releases CuFe2O4 and SRF in SRF@CuFe2O4@PEA81 NPs,and the release of CuFe2O4 can further accelerate the Fenton reaction.2.The results of cell uptake showed that SRF@CuFe2O4@PEA81 NPs could be endocytosis by Hepal-6 cells.CCK-8 assay showed that Hepal-6 cells exhibited good activity even at 200 g/mL concentration.After 808nm NIR laser irradiation,the*OH probe detection indicated that the intracellular Fenton reaction was enhanced.Calcein-AM/PI cells staining and flow cytometry showed that SRF@CuFe2O4@PEA81 NPS had a significant anti-HCC effect and promoted the apoptosis and necrosis of HEPA1-6 cells.3.The subcutaneous tumor model of Hepa1-6 cells was successfully constructed.8 h after intravenous administration SRF@CuFe2O4@PEA81 NPs,T2 signal strength and T2 value in the tumor region decreased to the lowest.The subcutaneous tumor volume and tumor weight of the SRF@CuFe2O4@PEA81+NIR group were the smallest,indicating that SRF@CuFe2O4@PEA81 had the strongest anti-HCC efficacy after being irradiated by 808nm laser.Compared with the control group,there was no significant difference in the body weight of mice in the treatment groups.Histopathological examination showed that SRF@CuFe2O4@PEA81+NIR group had a wide range of apoptosis and necrosis in tumor tissues,and its anti-HCC effect was the most significant.Conclusions:In this study,we successfully synthesized a multifunctional therapeutic integration reagent SRF@CuFe2O4@PEA81.This strategy combine real-time tracking,NIR-controlled drug release and,PTT and synergistic CDT,showing excellent anti-HCC efficacy.SRF@CuFe2O4@PEA81 NPs has ultra-sensitive T2-weighted MRI contrasting performance that are particularly useful for tumor-specific imaging.