Biocompatibility Assessment of TA-SeNPs

The in vitro biocompatibility of TA-SeNPs was investigated on fresh red blood cells (RBCs) as reported (Nagime et al., 2023 (link)). Briefly, TA-SeNPs, in a concentration range of 100–12.5 μg/mL, were incubated with an equal volume of RBCs. Phosphate buffer and Triton- X were tested as controls separately. After incubation for 1 h at 37°C, treated RBCs were centrifuged at 3,000 rpm for 5 min. The absorbance of the supernatant was measured at 540 nm using a microplate reader (UV/Vis-1800, Shimadzu, Kyoto, Japan). Furthermore, the cytocompatibility of HaCaT cells against TA-SeNPs was investigated using the mitochondrial MTT assay. Concisely, HaCaT cells were grown in a culture medium at a density of 1 × 10⁴ cells/well in 96-well plates as reported (Chidrawar et al., 2023 (link)). TA-SeNPs were incubated at a concentration range of 100–12.5 μg/mL for 24 h. After incubation, the spent medium was removed and treated with MTT (0.5 mg/mL) for 2 h, and the absorbance of DMSO-solubilized formazan crystals was measured at 560 nm using a microplate reader (UV/Vis-1800, Shimadzu, Kyoto, Japan).