Study on Multifunctional Nanoparticles Co-loaded with Indocyanine Green and Nrf2-siRNA against Oral Squamous Cell Carcinoma In Vitro

doi:10.13701/j.cnki.kqyxyj.2024.05.009

Abstract

Abstract: Objective: To construct multifunctional nanoparticles co-loaded with indocyanine green (ICG) and nuclear 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 carcinoma (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 distribution, zeta potential, and drug loading efficiency of nanoparticles were characterized. The SCC-25 cellular uptake of ICG and the escaping lysosomal phagocytosis ability of Nrf2-siRNA were evaluated. The photothermal and photodynamic 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-related 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 nanoparticles exhibited spherical structure with regular morphology and particle size of about 180 nm. PPI-siRNA nanoparticles 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 photodynamic properties. Nrf2-siRNA could significantly improve the anti-tumor efficiency of phototherapy by down-regulating 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.

Key words: oral squamous cell carcinoma, nanoparticles, photothermal/ photodynamic, gene silencing, antioxidant inhibition effect

FU Kaiyu, CHANG Yunhan, SHI Enyu, SHI Shurui. Study on Multifunctional Nanoparticles Co-loaded with Indocyanine Green and Nrf2-siRNA against Oral Squamous Cell Carcinoma In Vitro[J]. Journal of Oral Science Research, 2024, 40(5): 422-428.

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