Boosting Synergistic Effect Between Reactive Oxygen Species Based Therapy And Magnetic Hyperthermal Therapy In The Treatment Of Prostatic Cancer

Objective:As a promising treatment modality with spatial and temporal control,alternating magnetic field(AMF)triggered magnetic hyperthermal therapy(MHT)shows broad and promising applications to overcome the drawbacks of traditional focal therapies in combating cancer.However,the MHT efficacy is still not satisfactory with the presence of heat shock proteins(HSPs).Herein,we reported two new multifunctional reactive oxygen species(ROS)-related nanomedicine/ROS-based therapy activated through sonodynamic therapy(SDT)and chemodynamic therapy(CDT):(1)HP-HIONs@PDA-PEG and(2)HIONCs-GOD,based on nanobiotechnology for efficient cancer therapies.Methods:In this study,(1)HP-HIONs@PDA-PEG was at first designed for enhanced SDT and synthesized by a one-pot solvothermal process and sequential modification.A comprehensive characterization of nanohybrid was carried out.Catalase activity,SDT effect and heating capacity were performed.In vivo,subcutaneous implantation in nude mice were established and divided into six groups:the Control,PBS+US,HP+US,HP-HIONs@PDA-PEG+US,HP-HIONs@PDA-PEG+AMF and HP-HIONs@PDA-PEG+US+AMF.After different treatments,the anticancer effects and molecular mechanism of synergistic treatments were evaluated by comparison of tumor volume,immunofluorescence staining(hypoxia-inducible factor(HIF-1),HSP70 and HSP90),terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end labeling(TUNEL)staining,hematoxylin and eosin(HE)staining and immunohistochemical staining(Caspase 3 and Caspase 9);(2)HIONCs-GOD was designed for enhanced CDT and synthesized by a one-pot solvothermal process and the synthesized HIONCs were then etched with hydrochloric acid.A comprehensive characterization of nanohybrid was carried out.Catalase activity was then performed.The level of H₂O₂and p H value was measured to verify the effect of GOD in providing desirable environment for CDT.Then hydroxyl radicals(·OH)production test was performed to show the ability of nanoparticles in generating ROS.We then performed magnetic hyperthermia to show the capacity in heating.In vivo,subcutaneous implantation in nude mice were established and divided into seven groups:HIONCs-GOD+AMF,HIONCs+AMF,HIONCs,HIONCs-GOD,Free-GOD,PBS+AMF and the Control.After different treatments,the anticancer effects and molecular mechanism of synergistic treatments were evaluated by comparison of tumor volume,immunofluorescence staining(HSP70 and HSP90),TUNEL staining,HE staining and immunohistochemical staining(Caspase 3 and Caspase 9).Results:(1)HP-HIONs@PDA-PEG and(2)HIONCs-GOD were successfully synthesized.The results revealed that the hydrodynamic size of HP-HIONs@PDA-PEG and HIONCs-GOD were about 526 nm and 255 nm.The high crystallinity of HIONs/HIONCs guarantees their desirable magnetic properties.Oxygen related results demonstrated that our nanoparticles were appropriate candidates for self-supplying in situ oxygen generation and inherent hypoxia attenuation.ROS tests showed the ability of HP-HIONs@PDA-PEG and HIONCs-GOD in efficiently yielding highly toxic species either upon acoustic irradiation or in situ p H/H₂O₂slef-responsing.In vivo experiments,cleavage of HSP70 and HSP90 showed the good efficiency of nanoparticles for decreasing thermotolerance;TUNEL staining,HE staining and Caspase 3 and Caspase 9 staining indicated the enhanced therapeutic effects by SDT/MHT and CDT/MHT,independently.Above all,tests carried out on HP-HIONs@PDA-PEG and HIONCs-GOD nanohybrid aqueous dispersion demonstrated their effects on hypoxia attenuation and inducing reactive oxygen species,thus enhancing magnetic hyperthermal therapy.Conclusion:Thermotolerance can be significantly eliminated by either SDT or CDT,following by enhanced MHT.