Toxicological evaluation of ionic liquid in a biological functional tissue construct model based on nano-hydroxyapatite/chitosan/gelatin hybrid scaffolds.

The application of ionic liquid is attracting more attentions as green replacement for volatile organic solvents. However, the toxic effects and risks of ionic liquid in different biological systems for human health and environment are poorly evaluated. Among all ionic liquids, 1-ethyl-3-methylimidazolium diethylphosphate ([Emim]DEP-type) ionic liquid is still at the early phase of development, and its toxicity remains unclear. In this study, we fabricated a 3D biological functional tissue construct model based on nano-hydroxyapatite, chitosan and gelatin hybrid scaffold and evaluated its toxic effects of [Emim]DEP-type ionic liquid. As a control group, the examination of ionic liquid's toxic effects on the pre-osteoblast cell line (MC3T3-E1) was detected in 2D cultures. The MTT assay showed that [Emim]DEP-type ionic liquid inhibited the proliferation of cells on both 2D cultures and 3D tissue constructs. This effect was correlated with culturing time and concentration, while the IC50 on 3D scaffolds (12,566, 9015, 7896 μg/mL, at 24 h, 48 h and 72 h, respectively) was found significantly higher compared to 2D cultures (3959, 2226, 1884 μg/mL). Flow cytometry analysis and scanning electron microscope demonstrated that when [Emim]DEP-type ionic liquid acted on MC3T3-E1 cells for 48 h, the shape of 2D cells shrank, together with decreased surface adhesion.