Evaluation of Viability and Proliferation Profiles on Macrophages Treated with Silica Nanoparticles In Vitro via Plate-Based, Flow Cytometry, and Coulter Counter Assays
Figure 2
SiO2 NP effect on cell viability determined by plate-based and flow cytometry assays. RAW 264.7 mouse macrophage-like cells were cultured in DMEM-supplemented media for 48 h after which cells were exposed to SiO2 NP for 24 h, and viability was evaluated by plate-based assays (CellTiter-Blue, XTT, and LDH) and flow cytometry (PI staining and Calcein AM). (a) CellTiter-Blue and XTT assays determined increased values over control at low SiO2 NP concentrations. LDH-release, PI, Calcein AM, and LDH-release staining follow a concentration-dependent pattern with significant change at 0.1 g/L and 1 g/L SiO2 NPs, respectively. (b) CellTiter-Blue, PI, and Calcein AM staining show significant decrease in mitochondrial activity and viability, respectively, at 0.1 g/L and 1 g/L SiO2 NPs whereas XTT and LDH show significant change only at 1 g/L SiO2 NPs. Bars show average and SEM of 5–9 samples from independent experiments. Statistical analysis was performed using one-way ANOVA followed by Tukey’s multiple comparison test, and *() shows statistical significant difference versus control. # shows statistical significant difference in 0.1 g/L versus 0.01 g/L and 1 g/L versus 0.1 g/L, respectively, (CellTiter-Blue); $ shows statistical significant difference in 1 g/L versus 0.1 g/L (XTT); & shows statistical significant difference in 0.1 g/L versus 0.01 g/L (LDH); Δ shows statistical significant difference in 1 g/L versus 0.1 g/L (LDH release); α shows statistical significant difference in 1 g/L versus 0.1 g/L (PI and Calcein AM, resp.). Dashed line at 100 represents control cells viability.
