| The alginate-doxorubicin macromolecule prodrug vesicle(ALG-DOX-V)was prepared and characterized,and its anti-tumor effect was evaluated.The specific work content is as follows:Chemical preparation and characterization of physical and chemical properties of ALG-DOX-V: ALG-DOX macromolecular prodrug was synthesized by the condensation reaction of amino groups and substituted amides.Because the molecular structure of doxorubicin contains amino groups(-NH2),the molecular structure of alginic acid contains a large amount of carbonic acid(-COOH).EDC / NHS was used to activate-COOH on guluronic acid with high chemical activity in alginic acid structural units to obtain ALG-DOX macromolecular prodrug.The molecular prodrug self-assembles in solution due to the hydrophilicity to obtain the final product ALG-DOX-V.Transmission electron microscopy results showed that ALG-DOX-V had a hollow spherical vesicle structure.The average particle sizer of ALG-DOX-V detected by Malvern Zeta-Sizer nanoparticle size analyzer was 169.2 ± 3.19 nm,showing a normal single-peak distribution,and the Zeta potential was-31.7 ± 1.6 m V,and this nanosystem was stable in solution.The drug loading measured by ultraviolet spectrophotometry was 19.8 ± 1.92%,and the encapsulation replaced 67.3 ± 2.83%.The dynamic dialysis method was used to simulate the cumulative release rate of ALG-DOX-V at different p H values of human physiological conditions.The results displayed that the release process of ALG-DOX-V showed obvious slow release.In vitro biocompatibility evaluation of ALG-DOX-V: rabbit blood was used to detect the hemolysis rate of ALG-DOX-V and DOX;MTT colorimetric method was used to study the effect of DOX and ALG-DOX-V on HUVEC,L02 and H9C2 cells.The results show that ALG-DOX-V has better blood and cell compatibility than free DOX.Evaluation of the anti-tumor effect of ALG-DOX-V in vitro: The survival rates of BEL-7402,SMMC-7721 and Hep G2 hepatocellular carcinoma cells were determined by MTT colorimetry.The results showed that the cell survival rate was dose-dependent and time-dependent.The uptake of ALG-DOX-V by BEL-7402,SMMC-7721 and Hep G2 cells was analyzed by fluorescence photography.The results proved that the prolonged time of drug implantation into cells increased the cellular uptake of ALG-DOX-V gradually,which matched the results of cytotoxicity experiments.Flow cytometry was used to compare the uptake of DOX and ALG-DOX-V in BEL-7402 cells.The results showed that ALG-DOX-V had a certain sustained-release performance.Fluorescence spectroscopy and competitive binding experiments were used to determine the uptake of DOX and ALG-DOX-V in BEL-7402 cells.The results showed that ALG-DOX-V entered the cell through endocytosis mediated by mannose receptor.In vivo anti-tumor effect study of ALG-DOX-V: nude mouse xenograft model was established using BEL-7402,and the drug was injected every other day for 7 times.The tumor volume and weight of nude mice were recorded continuously.Tumor tissues and heart,liver and kidney tissues in each group were stained with H&E to evaluate the inhibitory effect of alg-dox-v on tumor proliferation and its toxicity to normal tissues.Compared with DOX,ALG-DOX-V stained tumors,H&E stained the heart,liver and kidney tissues of each group to evaluate the inhibitory effect of ALG-DOX-V on tumor growth and toxicity to normal tissues.Constantly reducing the inhibitory effect and effectively reducing the toxic and side effects of DOX on normal tissues. |
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