Synthesis Of CuS Core-shell Composite And Its Photothermal Killing Effect On Glioma

Objective As a new adjuvant therapy for tumor,photothermal therapy has become a research hotspot in tumor therapy in recent years with the advantages of strong targeting and minimally invasive.The synthesis of efficient and stable photothermal nanomaterials is particularly important for tumor photothermal therapy.In this study,Cu S core-shell composite(Cu S-PEG-FA)was synthesized by hydrothermal method to explore its killing effect on glioma cells,in order to provide a new theoretical basis for the application of photothermal nanomaterials in tumor photothermal therapyMethods 1.Copper chloride and sodium thiosulfate were used as copper and sulfur sources.Copper sulfide precursor nanoparticles were prepared by hydrothermal method at different temperatures.The most suitable temperature was selected according to the physical properties of copper sulfide;The materials were characterized by X-ray diffractometer,transmission electron microscope and Zeta potentiometer.2.Determine the photothermal conversion performance,and compare the photothermal conversion performance of Cu S-PEG-FA synthesized at different temperatures;The average survival rate of cells was measured by CCK-8 method Effect of Cu S-PEG-FA(6.25 μg/m L,12.5μg/m L,25 μg/m L,50 μg/m L,100 μg/m L)on the viability of human brain neuroblastoma cell line(LN229);the phagocytic effect of LN229 cells on Cu S-PEG-FA was observed by laser confocal microscope;The expression of phagocytosis associated protein caveolin-1(Ca V-1)was detected by Western blot and real-time PCR.Results 1 Through the identification of X-ray diffraction and physical properties of copper sulfide,it was found that Cu S NPs were successfully prepared by hydrothermal method at 50℃ and 70℃;2 Cu S-PEG-FA with larger particle size was prepared by coating Cu S NPs with PEG and FA.The particle size of Cu S NPs in the low-temperature synthesis group before coating was 25 nm,and the particle size of Cu S-PEG-FA in the low-temperature synthesis group after coating was 162 nm;The particle size of hightemperature combined group was 19 nm before coating,and that of Cu S-PEG-FA after coating was 116 nm.The particle size of low-temperature synthesis group was larger than that of high-temperature synthesis group before and after coating;When the illumination time reached 5 min,the average temperatures of Cu S-PEG-FA in each dose group of low-temperature synthesis group were 25.9℃,28.7℃,31.5℃,38.7℃ and 43.3℃respectively,while those in each dose group of Cu S-PEG-FA in high-temperature synthesis group were 25.8℃,29.9℃,31.5℃,39.2℃ and 44.2℃ respectively.The experimental results showed that the temperature of Cu S-PEG-FA in high-temperature synthesis group reached 44.2±1.0℃.after 5 min illumination,which was higher than43.3±1.2℃ in low-temperature synthesis group,The results showed that the photothermal conversion performance of Cu S-PEG-FA synthesized at high temperature was better than that synthesized at low temperature;The results of CCK-8 showed that the average survival rates of LN229 cells in the low-temperature synthesis group were 77.61%,75.90%,62.57%,60.65% and 23.93% respectively after exposure to near-infrared light with the wavelength of 1064 nm for 5 minutes,while the average survival rates of cells in each concentration group of Cu S-PEG-FA in the high-temperature synthesis group were73.82%,71.92%,56.90%,53.58% and 15.31% respectively.The killing effect of the high-temperature synthesis group on LN229 cells was better than that of the lowtemperature synthesis group.The concentration of Cu S-PEG-FA in the high-temperature synthesis group is 100 μg/m L had the best killing effect on LN229 cells;The results of cell phagocytosis showed that the phagocytic effect of LN229 cells on Cu S-PEG-FA increased with the extension of incubation time;After different concentrations of Cu SPEG-FA acted on LN229 cells,the expression of Ca V-1 protein and m RNA increased of which 25 μg/Ml,50 μg/Ml,100 μg/m L group,the expression of Ca V-1 in increased by1.11,1.12 and 3.62 times respectively;25 μg/m L,50 μg/m L;100 μg/m L;The expression of Ca V-1 m RNA in group was 1.32,1.41 and 3.21 times higher than that in the control group,respectively.Conclusion Cu S NPs were successfully prepared by hydrothermal method at 50℃ and70℃;Cu S-PEG-FA prepared after coating PEG and FA shell can produce photothermal killing effect on LN229 cells under near-infrared light,but it has no significant effect on the cycle of LN229 cells;The phagocytic effect of LN229 cells on Cu S-PEG-FA could be increased by prolonging the incubation time;The increased expression of phagocytic protein Ca V-1 suggests that phagocytic protein Ca V-1 may be involved in the process of Cu S-PEG-FA entering LN229 cells.Figure9;Table4;Reference 135...