| Objective:Photothermal therapy(PTT),as an alternative or supplement to traditional tumor treatment methods,has the advantages of remote manipulation,low invasion,and high safety.It is a rapidly developing tumor treatment technology.Although many different nanomaterials have achieved good therapeutic effects in the application of PTT,due to poor aggregation at tumor sites and high tumor heat resistance,PTT has low efficacy and frequent tumor recurrence.Therefore,this study designed a new multifunctional nanomaterial to study its anti-tumor effect in the field of PTT,providing a new strategy for reducing the side effects caused by high temperature.Methods:1.Polyamine nanoparticles(PDA NPs)were synthesized by oxidative self-polymerization of dopamine hydrochloride monomer.Hyaluronic acid(HA)and PDA NPs were covalently bound through an amidation reaction.The heat shock protein 90(HSP90)inhibitor Ganetespib(STA9090)was loaded onto PDA NPs through aπ-πstacking mode of action.Subsequently,the synthesized NPs were characterized by scanning electron microscopy(SEM),transmission electron microscopy(TEM),nano laser particle size analyzer,UV,and H~1NMR;At the same time,an 808 nm near infrared laser was used to irradiate different concentrations of NPs aqueous solutions,and thermocouples were used to monitor temperature changes and photothermal stability during the illumination process.2.The expression of CD44 protein in A549 and BEAS-2B was detected by real-time fluorescence quantitative polymerase chain reaction(real-time PCR).Using a laser confocal microscope,the cell uptake effect of NPs was analyzed,and then the survival and apoptosis rates of A549 and BEAS-2B cells under different treatment conditions were measured using different detection kits.Western Blot was used to detect the changes in HSP90 expression level of A549 cells in different treatment groups.3.Different NPs solutions were injected into the tail vein of mice to observe the photothermal effects of NPs in vivo.After 14 days of treatment,the therapeutic efficacy and biocompatibility of NPs in vivo were investigated by measuring the tumor volume and weight,as well as the body weight of mice during treatment.Tumor tissue sections were stained with hematoxylin eosin(H&E).Results:1.The synthesized NPs were observed to be spherical by SEM and TEM.The average particle size of PDA NPs measured by the nano laser particle size analyzer after reacting with HA increased from 139.86±1.00 nm to 180.57±1.6 nm.After loading STA9090,the particle size of PDA-HA/STA9090 NPs increased to 209.94±1.57 nm.The characteristic UV absorption peak and characteristic hydrogen chemical shift of STA9090were observed on the UV spectrum and nuclear magnetic resonance hydrogen spectrum of PDA-HA/STA9090 NPs.The temperature rise measurement results show that PDA-HA/STA9090 NPs have a good photothermal effect at a concentration of 200μg/m L when the light power is 1.6 W/cm~2 and the solution temperature reaches 49.3℃,the photothermal conversion efficiency is calculated to be 35.2%.2.PCR results showed that the expression of CD44 protein in tumor cells was 1.5 times higher than that in normal cells.The uptake of PDA-HA and PDA-HA/STA9090 NPs by cells was greater than that of PDA NPs.The results of enzyme labeling and laser confocal microscopy showed that PDA-HA/STA9090 NPs had a significant tumor killing effect,with a cell apoptosis rate of78.3%measured by flow cytometry.Western Blot results showed that the expression of HSP90 in the PDA-HA/STA9090 group was lower than that in the control group with or without light.3.After 4 hours of injection of NPs solution,the tumor site was irradiated with 808 nm laser,and its surface temperature rapidly increased to 50℃.During the treatment period,the tumor volume in the control group rapidly increased,while the tumor volume in the PDA-HA/STA9090 light irradiation group significantly decreased compared to the control group.After 14 days,the tumor weight in the PDA-HA/STA9090 light group was the lightest.In addition,the H&E staining results of tumor tissue sections showed that the tumor cells in the PDA-HA/STA9090 light group were necrotic in large numbers,and the purple nucleoli were reduced.Conclusion:1.The prepared PDA,PDA-HA,and PDA-HA/STA9090 NPs have uniform particle sizes,good particle size stability,and photothermal effects.The successful synthesis of composite nanomaterials and their excellent photothermal properties provide a basic basis for the next step in vitro cytological experiments and in vivo anti-tumor evaluation.2.This experiment evaluated the tumor cytotoxicity of PDA-HA/STA9090 NPs through in vitro cytology,further providing an important basis for subsequent in vivo anti-tumor experiments.In addition,the WB experiment results verified the inhibitory effect of the HSP90 inhibitor STA9090 on the expression of HSP90 in cells,providing a research basis for the application of STA9090 in the field of PTT.3.In vivo animal experiments have shown that PDA-HA/STA9090 NPs exhibit good photothermal effects in vivo.The results of tumor volume and weight during treatment confirmed that PDA-HA/STA9090NPs had a good tumor treatment effect.In summary,the novel multifunctional nanomaterials synthesized exhibit synergistic therapeutic effects of PTT and HSP inhibition in vitro and in vivo,and this strategy of improving the efficacy of PTT through synergistic tumor targeting and heat shock protein inhibition has good application prospects. |
