Preparation Of Selenium-loading Inorganic Nanoparticles Modified By Small Organic Molecules And Its Anti-tumor Research In Vitro

Objective:A lot of papers have shown that inorganic selenium ions have obvious anti-tumor effects,but they are not easy to control due to dosage,especially when they are exposed to high doses,which limits their clinical application.This study intends to use small organic molecules of phytic acid?IH?as morphology control agent,the preparation of a new type of p H sensitivity,low toxicity and selenium-loading inorganic nanoparticles can controlled the release of selenium ions,inorganic selenium so as to provide an experimental basis for clinical application of ion and theoretical basis.Methods:?1?In order to study the effect of IH content on the growth of nanoparticles,two methods were added to different amounts of IH,followed by 0,0.33%?v/v?,1%?v/v?and 1.67%?v/v?respectively.Diffusion method:centrifugal tube,the centrifugal tube foil cover,after punching in a closed beaker filled with ammonia,room temperature for 12 hours;hydrothermal method:high pressure reaction vessel,120?,3 hours.?2?Characterization of selenium-loading compounds by SEM,XRD,FTIR,XPS,TG-DTA andmorphology;?3?PBS were adjusted to pH=7.4 and pH=5.8,to detect the degradation of nano particle accumulation release situation by detecting the concentration of Sein solution;?4?Eca109 cells were cultured in vitro,using sodium selenite and selenium nanoparticles synthesized to interfere,growth curve,cell proliferation was detected using CCK-8 method.The cell morphology was observed by Wright staining,cell cycle,apoptosis,lactic acid secretion index;?5?statistical analysis by SPSS.Results:?1?Under the condition of normal temperature and pressure,different powder like samples were successfully obtained by diffusion of ammonia water and the increase of pH in solution.The results of SEM electron microscopy showed that the product was even thin when the IH was not added.After joining IH,the product sheet gradually transformed into nano particle size and smooth surface;XRD results showed that the product was CaH₂O₄Se;FTIR results show that the product contains water of crystallization and SeO₃^2-,and increased with the increasing amount of IH,were also detected by PO₄^3-;XPS analysis showed that the product contains Ca,Se,O,C and P,where Se is the+4 valence;TG-DTA analysis shows that the product is decomposed under high temperature.?2?Under the hydrothermal condition,different powder like samples were successfully obtained by the decomposition of urea and the increase of pH in the solution.The results of SEM showed that SEM without IH product is thin,with the increasing amount of IH,the product transferred from flake to nanoparticles smooth,then transformed into surface with acicular nano structure of XRD microspheres;the results showed that when the IH content is less than or equal to 1%?v/v?,the product is CaH₂O₄Se,when the amount of IH 1.67%?v/v?,hydroxyapatite in the product?Ca₅?PO₄?₃?OH??.And with the increasing amount of IH,reduce the degree of crystallinity,transformation of amorphous;FTIR results showed that the main product contains crystal water and SeO₃^2-,and increased with the increasing amount of IH,detected by PO₄^3-;XPS results showed that Se,O,containing Ca,C and P elements in the product,where Se is the valence of+4;TG-DTA results show that at high temperature and decomposition products.?3?The experiment of ion release was carried out by the nanoparticles prepared under the condition of normal temperature and hydrothermal condition.The results show that the release of selenium ions is faster under the neutral condition than in the acidic condition.Under the condition of acid,the release of selenium ions from nanoscale slices at normal temperature is faster than that of nano thin slices prepared under the condition of water and heat without adding IH.After adding IH,it can effectively delay the release of selenium.?4?Inorganic selenium nanoparticles co cultured with tumor cells results:1.Results:CCK-8 showed that without adding the product of IH after co cultured with tumor cells,no cell proliferation,and adding sodium selenite in the medium of cell proliferation were similar;when the dosage of IH was 0.33%?v/v?,co culture products and cell,cell proliferation was not obvious;when the amount of IH is more than or equal to 1%?v/v?,co culture products and cell,the number of cells showed a decreasing trend after the first increase;the Wright's staining showed that tumor cell membrane shrinkage,cytoplasm structure chaos,pyknosis,nuclear membrane clear display;apoptosis detection,the high concentration group?S2?in cell death accounted for 21.28%,late apoptosis accounted for 19.22%,accounting for 1.03%of early apoptosis.The late apoptotic cells in the low concentration group?S1?accounted for 1.39%,and the late apoptotic cells in the control group accounted for 0.06%.The results of cell cycle detection showed that G2/G1 in group S1,group S2 and group control were 1.94,1.93,1.91,respectively.The proportion of G1,S and G2was almost the same.The results of S1 and S2 were similar in the experimental group.Conclusion:1.At room temperature and under hydrothermal conditions,the morphology and crystallinity of organic small molecule IH can regulate inorganic nano particles,thereby affecting the degradation rate and ion release rate of nanoparticles;2.By IH regulation generated with pH sensitivity,the degradation rate under acidic conditions was faster than that under neutral condition.Moreover,the release of selenium can be controlled by adjusting the concentration of IH.The results of co culture of;3.in vitro cultured cells can produce acidic substances in the process of proliferation,degradation of these acids can accelerate the inorganic selenium nanoparticles;4.selenium nanoparticles and tumor cells co culture results showed that,by adding the amount of adjustment of IH,can control the products in vitro degradation,control of inorganic selenium ions produced thereby has the effect of inhibiting the growth of tumor cells,and reduce the toxicity of inorganic selenium ions;5.inorganic selenium ions can effectively inhibit the proliferation of tumor cells,but the specific mechanism needs further study;...