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The performance of quasi-spherical gold nanoparticles (GNPs) on the generation of reactive oxygen species (ROS) to cause cell damage, as irradiated by a two-photon laser, is studied. In this mechanism, hot electrons are generated from GNPs as irradiated by the two-photon laser, reacting with the molecules in the medium to produce ROS. We used laser scanning confocal microscopy with a low-fluence femtosecond Ti:Sapphire laser of 800 nm to observe the generated ROS in A431 cells, which were incubated with GNPs in advance. Subsequently, the cell morphology, cytoskeleton, and viability were investigated. In comparison with the control (no GNPs), the expression of ROS in these GNP-treated cells was enhanced after irradiation by the two-photon laser. Additionally, the disruption of cytoskeletons and the follow-up apoptosis of these GNP-treated cells are significantly increased as the number of laser shots increases. Moreover, we used N-acetyl-L-cysteine (NAC), an antioxidant, to inhibit the formation of ROS, to clarify whether the cytoskeletal disruption is caused by ROS rather than photothermal effects. Our results show that after two-photon irradiation, the ROS expression in these cells treated with GNPs plus NAC was significantly reduced. In addition, the cytoskeletal damage of these cells treated with GNPs and NAC was less than that of those treated with GNPs but without NAC; their cell viability after three days was almost the same with the control. These results illustrate that the induced ROS from the two-photon excited GNPs is the main cause of cell damage. The study may pave a way for the use of GNPs as a photosensitized therapeutic agent for two-photon photodynamic therapy on tumor treatment.

The Effect of Quasi-Spherical Gold Nanoparticles on Two-Photon Induced Reactive Oxygen Species for Cell Damage