Figure 3. Toxicity assay of Mn₃O₄ NPs in vitro and in vivo.

(a–b) The data of cell viability on CNE-2, NP69 and 293 cells incubated with different concentrations (150 μM, 100 μM, and 10 μM) of the Mn₃O₄ NPs for 24 and 48 h. (c–d) Cell death and Apoptosis rate of CNE-2 and NP69 cells were measured by flow cytometry at 48 h after incubation of PBS, LPS, Mn₃O₄ NPs (150 μM and 200 μM). Cells were stained by annexin V and PI. (e) Cells absorption data of the Mn₃O₄ NPs. TEM images of CNE-2 and NP69 at 12 h after incubation with the Mn₃O₄ NPs (100 mmol/L). (f) Immunotoxicity assay in vivo. CD80, CD86, CD11b and CD206 expression of monocytes/macrophages in peripheral blood, as well as CD69 cytokine of adaptive immune in lymphocyte cells of peripheral blood (BL) and lymph nodes (LN). * P < 0.05 compared with Gd-DTPA group.