| BackgroundContinuously updated diagnostic methods and advanced imaging methods have led to an increase in the early detection rate of small liver cancer,however,even with current diagnosis methods,it is still challenging to accurately judge a nodule with a diameter less than 2 cm whether it is hepatocellular carcinoma or liver cirrhosis.To solve this issue,a new technology is needed to distinguish above two kinds of liver nodules.There is an emerging imaging method that improves tissue resolution and sensitivity to detect micro-nodules with diameters less than 1 cm.To detect micro-nodules,photoacoustic imaging(PAI)was used to provide normnvasive images at depths of several centimeters with a resolution of approximately 100 μm.To improve specificity,we developed a probe that specifically targets hepatocellular carcinoma by recognizing the biomarker GPC3 on the hepatocellular carcinoma cell membrane.The probe not only has a strong photoacoustic signal,but also have magnetic resonance signal.Furthermore,the material own photothermal effect which absorbs longer wavelength light and releases heat,effectively and accurately kills tumor cells,thus improving patient survival and postoperative quality of life.Herein,we present a new technology that uses photoacoustic imaging to image and target micro-hepatocellular carcinoma biological processes derived from liver cirrhosis with high spatial resolution.Objective1.Identification:Identification between micro-hepatocellular carcinoma and liver cirrhosis by specifically targeting hepatocellular carcinoma cell membrane of targeted probe.2.Treatment:The photothermal properties of targeted probes was used to treat micro-hepatocellular carcinoma accurately.Method1.Synthesis and Characterization of nanosheet Fese2.The Synthesis of nanosheet Fese2 was made by the Oleic acid method.The hydrodynamic diameters and zeta potentials of Fese2 were measured by Zetasizer Nano(ZS),the images and diameters were measured by transmission electron microscopy(TEM),absorbance spectrum was measured by UV-3600Plus spectrophotometer.2.Surface modification of nanosheet FeSe2 and Characterization of nanosheet FeSe2-PEG-Peptide.Nanosheet FeSe2 was modified by PEG,and then by Peptide.The hydrodynamic diameters and zeta potentials of FeSe2 were measured by Zetasizer Nano(ZS),the images and diameters were measured by transmission electron microscopy(TEM),absorbance spectrum was measured by UV-3600Plus spectrophotometer.3.Cell toxicity test of targeted probe FeSe2-PEG-Peptide,targeting ability of probe at the cellular level,Photothermal effect of probe in vitro.The MTS assay was used to evaluate cell toxicity of targeted probe.Tissue perfusion and Transmission electron microscope was used to evaluate ability of probe.Irradiated with different energy lasers and then stained with live cells by AM/PI was used to evaluate photothermal effect at the cellular level.4.Five-week-old male Balb/c nude mice were used to establish an orthotopic liver cancer model and 5-week-old male Balb/c mices were used to establish a liver cirrhosis model5.The targeting ability of targeted nano-probe FeSe2-PEG-Peptide at the animal level.Then,probe metabolic of above three groups mice were measured by photoacoustic imaging equipment.The comparison of different mice groups can reflect the targeting ability of FeSe2-PEG-Peptide at the animal level.6.Photothermal therapy and verification of therapeutic effect.Three groups of mice were then weighed,autofluorescence imaging detection,and survival time every 5 days.Finally,the liver of each mouse was taken for pathology to test the therapeutic effect.ResultFor diagnosis,the metabolism of targeted probe in mice with situ liver cancer is faster than the non-targeted probe,and a high concentration of aggregation was around the tumor region.At the same time,the metabolism of targeted probes are slow in liver cirrhotic mice model,and there is almost non-aggregated around the liver cirrhosis nodules.Because of the significant difference,the two models could be identified absolutely.For treatment,after photothermotherapy in mice injected with PBS,the tumor did not decrease in size;After photothermotherapy in mice injected with non-targeted probe,the tumor first decreased and then increased in size,because tumor cells could not completely be killed and tumor was relapsed;After the photothermal treatment,the mice injected with targeted probe,the tumor cells were completely killed and tumor was cured.ConclusionWe have developed a novel targeting nanoprobe FeSe2-PEG-Peptide,combined with the advantages of dual-mode PA/MR imaging,to achieve the purpose of differential diagnosis of micro-hepatocellular carcinoma and cirrhosis.At last,the micro-hepatocellular carcinomas are stifled in the cradle by targeted photothermal therapy. |
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