Multimodal Imaging Nanoparticles For Predicting Prognosis And Precise Embolization Of Hepatocellular Carcinoma

Objective:hepatocellular carcinoma（HCC）is a common cancer.About 70%of HCC cases are unresectable at the time of discovery.Selective hepatic artery embolization is recommended as the main treatment for unresectable HCC.However,due to the failure of TACE to embolize tumor vessels accurately and the different response of patients to the efficacy of TACE,the overall survival benefit is not obvious.In order to improve the effect of tumor embolization,it is urgent to accurately embolize tumor vessels and screen suitable patients for embolization.At the same time,according to the relevant literature,we found that the higher the GSH level in tumor,the stronger the drug resistance and invasion of tumor.Therefore,imaging GSH of tumor will help to further understand the prognosis of tumor.Methods:in this study,we prepared MnO₂ nanosheets from MnCl₂,TMA·oh and H₂O₂.Further,we added Verteporfin（BPD）to manganese dioxide（MnO₂）solution,stirred vigorously for 2 h at room temperature,and then let it stand for 6 h.BPD can be loaded on MnO₂ surface.Free BPD was removed by 30 k Da Amicon（MWCO）ultrafiltration centrifuge.With the help of ultrasound,the obtained MnO₂/BPD was redispersed in water to obtain nano MnO₂/BPD solution.The size,shape,and chemical properties of MnO₂,MnO₂/BPD,and Nano BPD were tested by means of electron microscopy,hydration particle size,and in vitro reduction characteristics.The imaging performance was verified by measuring the changes of photoacoustic imaging,MR imaging and fluorescence imaging signals with different concentrations.In the aspect of targeting MnO₂/BPD,we verified the expression of LDLR,the target of BPD,in mice and human surgical specimens.Furthermore,we co stained the vessels with LDLR and found that LDLR was highly expressed in the tumor vessels.Finally,we injected nanoparticles into the body to see if the nanoparticles could target the blood vessels of the tumor.We divided the experimental group into 5 groups,namely,saline,MnO₂/BPD,saline+laser,BPD+laser,MnO₂/BPD+laser.We used the puncture needle to puncture the tumor under the guidance of ultrasound.After the puncture needle reached the tumor,we withdrew the needle core and introduced the optical fiber.Under the condition of infrared temperature measuring instrument,photodynamic irradiation was carried out for 8 min.In order to verify the effect of photodynamic therapy,we measured ROS,thrombus formation and calculated the embolism rate of blood vessels after photodynamic therapy.At the same time,in order to further understand the effect of interventional photodynamic therapy on tumor size and survival time of mice,we measured the tumor size change with IVIS every5 days,measured the weight change of mice every 3 days,and recorded the survival time of mice with monitoring during the treatment.In terms of imaging,based on good in vitro imaging,we further carried out multimodal imaging in vivo,Since nanodrugs are targeted to the blood vessels of tumor,we resected the tumor after multimodal imaging,stained the blood vessels with immunofluorescence,and calculated the relationship between the imaging intensity and the vascular density of tumor.Finally,the relationship between imaging intensity and embolization effect was explored.In order to further study the prognosis of HCC,we designed a GSH sensing nanoprobe MnO₂/ICG.It can sense the GSH content in tumor.Similarly,we measured the size,shape,chemical properties,and imaging ability of nanomaterials.After exploring the best imaging time point,the levels of GSH in different types of tumors were imaged.And it is associated with tumor invasion and drug resistance.Results:we successfully synthesized MnO₂/BPD,which had good dispersion and uniform size.MnO₂ in MnO₂/BPD can be reduced in GSH/H₂O₂ to obtain Mn²⁺.Mn²⁺can self-assemble with BPD to form Nano BPD.Nano BPD has enhanced photodynamic effect and good multimodal imaging effect.MnO₂/BPD can target tumor blood vessels.After photodynamic therapy,MnO₂/BPD can produce a lot of ROS and induce apoptosis of vascular endothelial cells.The apoptosis of vascular endothelial cells can trigger the coagulation cascade in local blood vessels,as to achieve accurate targeted embolization of blood vessels.In the MnO₂/BPD+laser group,the tumor size decreased significantly and the survival time prolonged significantly.At the same time,in terms of imaging,we found that MnO₂/BPD can image tumor vascular density and predict the prognosis of tumor embolization.The higher the imaging intensity of the tumor,the higher the vascular density of the tumor,and the better the embolization effect of the tumor.On the other hand,MnO₂/ICG can predict tumor invasion and drug resistance by sensing GSH concentration in tumor.The higher the GSH in the tumor,the lower the imaging intensity of MnO₂/ICG,the higher the invasiveness and drug resistance of the tumor,and the worse the prognosis of the tumorConclusion:the self-assembled nanoparticles were constructed,which greatly improved the photodynamic effect of photosensitizer.Nanoparticles can trigger coagulation cascade and embolize tumor accurately under the effect of interventional photodynamic therapy.Nanoparticles can target tumor neovascularization and image tumor vascular density.For the first time,the imaging intensity of tumor was used to evaluate the effect of tumor embolization.In this study,we constructed a multimodal imaging nanoparticle with GSH response to evaluate the GSH level in tumor.For the first time,GSH imaging intensity of nanoparticles was used to evaluate the prognosis of tumor.