| Objective:This study is divided into three sections:Ⅰ.the synthesis of Fe2O3-PDA-Dox nanoparticles and the verification of its physical and chemical properties;Ⅱ.the application of Fe2O3-PDA-Dox nanoparticles through the hepatic artery by interventional method to verify the TACE combination Efficacy and safety of PTA in the treatment of liver cancer in rats;Ⅲ.CA4P combined with Fe2O3-PDA-Dox nanoparticles in the treatment of liver cancer in rats,to verify whether CA4P can increase the uptake of Fe2O3-PDA-Dox nanoparticles by rat liver cancer tissue to improve TACE combination The efficacy of PTA.Materials and Methods:After synthesizing Fe2O3-PDA nanoparticles,the morphology and particle size were observed and detected by scanning electron microscopy and dynamic light scattering.The Dox release profile was measured and plotted by an ultraviolet-visible spectrophotometer under different pH conditions.The photothermal conversion efficiency of different concentrations of nanoparticles in vitro was monitored under a thermal imager.SD rats after 1 week of tumor-bearing were randomly divided into 5 groups,5 in each group,divided into pre-test group,control group,cTACE group(Lipidol+Dox),and dTACE group(Lipidol+Fe2O3-PDA-Dox),dTACE+PTA group(Lipidol+Fe2O3-PDA-Dox+NIR),CA4P group(Lipidol+Fe2O3-PDA-Dox+NIR+CA4P).The 3.0T MR scan was used to observe the characteristics of nano-particle T2 sequence images in the pre-experiment group and in vitro,and the presence of nanoparticles in tumor tissues was confirmed by Prussian blue staining.The release of Dox in tumor tissues was observed by fluorescence microscopy before surgery,1 hour after surgery,6 hours after surgery and 24 hours after surgery.The modified gastroduodenal artery was retrogradely intubated into the rat hepatic artery.The liver MRI and tail vein blood were taken before operation and on the 3rd,7th,and 10th day after surgery.Size changes and changes in liver and kidney function.On the 10th day after operation,the liver tumor tissues were examined by HE,TUNEL staining and Ki67,Caspase3,CD31 and VEGF immunohistochemistry.The CA2P group and dTACE+PTA group were scanned for T2 sequence at 1h and 24h after operation.Tumor tissue neutron activation analysis of tissue Fe content.Statistical analysis was performed using SPSS 23.0 software:quantitative data was expressed as x±s.The t test was used for comparison between the two groups.One-way analysis of variance was used for comparison between groups.The test level was a=0.05,P<0.05 was considered statistically significant.Results:The particle size of Fe2O3-PDA nanoparticles is about 100nm,and the thickness of PDA is about 10nm.The size is relatively uniform.UV-Vis spectra showed that Fe2O3-PDA-Dox and Dox showed similar characteristic peaks at about 478 nm,indicating that Fe2O3-PDA successfully loaded Dox molecules.The Dox release profile can be seen in an acidic(pH 6.5)environment with a higher Dox release rate.The temperature rise curve shows that Fe2O3-PDA-Dox has a good photothermal effect in vivo and in the outside,and the concentration is 20μg/ml.MR examination showed that Fe2O3-PDA-Dox was superparamagnetic,and the T2 signal was significantly lower than that of the control group.Prussian blue staining showed that Fe2O3-PDA-Dox accumulated in the tumor tissue.After Fe2O3-PDA-Dox was applied to the hepatic artery at different time points in the tumor tissue,the biodistribution results in the tumor tissue showed that Dox gradually released over time and had a good sustained release effect.There was a statistical difference in tumor volume changes between groups.The average content of Fe in the tumor tissue of CA4P group was 23.72±12.45 ug/g and 14.61±8.23 ug/g at 1h and 24h,respectively.The average content of Fe in tumor tissue at 1h and 24h after operation in dTACE+PTA group was 5.66±.4.29 ug/g,2.76±1.33 ug/g.The MRI T2 signal of the tumor in the CA4P group was lower than that of the dTACE+PTA group at 1h and 24h after surgery.The temperature of the tumor site increased significantly after injection of nanoparticles in the two groups.The dTACE+PTA group reached 55℃ in 5 minutes,while the CA4P group reached 62℃.Pathological and immunohistochemical showed tumor cell necrosis,apoptosis and proliferation inhibition after treatment,and the CA4P group was more significant-The two groups showed transient liver and kidney toxicity at 3 days,and the CA4P+pTACE group had slightly higher damage than the pTACE group,and gradually decreased on the 7th and 10th day.Conclusions:This study clarified the physicochemical properties of Fe2O3-PDA-DOX nanoparticles,which have:1.Small volume,high drug loading rate,easy to penetrate blood vessels into tumor cells;2.Release DOX in the acidic microenvironment of tumor tissue;3.Fe2O3-PDA has good photothermal conversion performance;4.Fe203-PDA has MRI T2 visual image features.Rats were treated with hepatic artery via interventional methods,and it was verified that Fe2O3-PDA-DOX nanoparticles can simultaneously have the effect of TACE+PTA.This study also confirmed that CA4P can increase the uptake of Fe2O3-PDA-DOX nanoparticles by rat hepatocellular carcinoma,which can enhance the combined treatment of transcatheter arterial chemoembolization and photothermal ablation without additional increase in liver and kidney toxicity.Safety is worthy of recognition,providing new methods and ideas for the treatment of liver cancer. |
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