The present study aims to evaluate the toxicity of the green calcium oxide nanoparticles (CaO-NPs) from golden linseed extract (Linum usitatissimum L.) by phytotoxicity in seeds (Daucus carota, Beet shankar, Lactuca sativa and Brassica oleracea), in vitro safety profile and soil toxicity for CaO-NPs solutions from 12.5 to 100 μg mL−1. Ecotoxicity analysis of the soil was conducted using XRD diffractograms, which revealed characteristic peaks of the nanoparticles at 37.35° (12.5, 25, 50, and 100 μg mL−1), as well as a peak at 67.34° (25 and 100 μg mL−1). Additionally, the in vitro safety assessment indicated favorable cell specification and regulation within the first 24 h, demonstrating reductions of 15.9 ± 0.2%, 17.9 ± 0.2%, 17.6 ± 0.2%, and 32.9 ± 0.2% to 12.5, 25, 50, and 100 μg mL−1, respectively. The dsDNA assay revealed initial protection and controlled release within the cells for 48 h. However, after 72 h, there was an increase of 20 ± 0.2%, 16 ± 0.2%, 32 ± 0.2%, and 43 ± 0.2% to 12.5, 25, and 50 μg mL−1. The analysis of ROS generation demonstrated a reduction of 40 ± 0.2%, 33 ± 0.2%, 20 ± 0.2%, and 9 ± 0.2% to 12.5, 25, 50, and 100 μg mL−1, respectively, within 72 h. When compared to the negative control (NC), there was an increase of 50 ± 0.2%, 56 ± 0.2%, 77 ± 0.2%, and 92 ± 0.2% at the same concentrations, suggesting that the nanoparticles generated free radicals, leading to cellular inflammation. This was attributed to the positive surface charge of the nanoparticles, resulting in reduced interaction with the cell membrane and the subsequent release of hydroxyl (•OH), which caused inflammatory processes in the cells. Therefore, CaO-NPs exhibited a low phytotoxicity and high cytocompatibility, while also promoting plant germination and growth.
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