Study on Multifunctional Nanoparticles Co-loaded with Indocyanine Green and Nrf2-siRNA against Oral Squamous Cell Carcinoma In Vitro
Objective:To construct multifunctional nanoparticles co-loaded with indocyanine green(ICG)and nu-clear factor erythroid 2-related factor 2-small interfering RNA(Nrf2-siRNA),and to explore the combined effects and mechanisms of photothermal and antioxidant inhibition amplified photodynamics against oral squamous cell car-cinoma(OSCC)in vitro.Methods:Double emulsion method and electrostatic adsorption were used to prepare PPI-siRNA nanoparticles based on polyurethane(PBAE)/polylactic acid-glycolic acid copolymer(PLGA)co-loaded with ICG and Nrf2-iRNA.The morphology,size distribu-tion,zeta potential,and drug loading efficiency of nanoparti-cles were characterized.The SCC-25 cellular uptake of ICG and the escaping lysosomal phagocytosis ability of Nrf2-siRNA were evaluated.The photothermal and photodynam-ic effects of PPI-siRNA nanoparticles were investigated by detecting the heating effect,expression of intracellular heat shock protein(HSP60),and the generation of reactive oxygen species(ROS).The expression levels of E2-re-lated factor 2(Nrf2)and its downstream genes GCLM and GCLC regulated by the Nrf2 were investigated to explore the assisting photodynamic effects mechanism of Nrf2-siRNA.The cytotoxic effect of PPI-siRNA nanoparticles combined with laser irradiation was evaluated by MTT assay and live/dead cell assay.Results:The PPI-siRNA nan-oparticles exhibited spherical structure with regular morphology and particle size of about 180 nm.PPI-siRNA nano-particles could efficiently deliver ICG and Nrf2-siRNA into cells,and prevent Nrf2-siRNA from being engulfing from lysosome to exert gene silence effect.PPI-siRNA nanoparticles possessed notable photothermal and photody-namic properties.Nrf2-siRNA could significantly improve the anti-tumor efficiency of phototherapy by down-regula-ting the expression of antioxidant proteins and genes.Conclusion:PPI-siRNA nanoparticles can effectively inhibit the in vitro growth of OSCC by combining phototherapy and gene silencing amplified photodynamic efficacy,and have a broad application prospect in the clinical treatment of OSCC.
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