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. 2020 Feb 21;8:100. doi: 10.3389/fbioe.2020.00100

Figure 5.

Figure 5

Cell viability studies in PBMCs and assay interference evaluation. (A) Cell viability decrease induced by different concentrations of Chit 80% NPs, Chit 93% NPs, Chit 80%, and Chit 93% in human PBMCs, evaluated by MTT assay following 24 h of incubation. Dotted line represents the 70% of cell viability. Results are expressed as mean ± SEM, obtained from four independent experiments, each performed in triplicate (n = 4). (B) Confirmation of MTT results by testing four different concentrations of Chit 80% NPs and Chit 93% NPs by flow cytometry using PI. Results are expressed as mean ± SEM, obtained from 1 to 4 independent experiments, each performed in duplicate (n = 1–4). (C) Nonlinear regression analysis of the cell viability data, allowing the extrapolation of IC50 values (720 μg/mL for Chit 80% NPs and 2104 μg/mL for Chit 93% NPs). Significant statistical difference between Chit 80% NPs IC50 and Chit 93% NPs IC50 calculated using extra sum-of-squares F-test. (D) Evaluation of possible NP and polymer interference with the wavelength used to read MTT assay (540/630 nm) (Mean ± SEM, n = 3). (E) Evaluation of the cell viability resultant from the incubation of PBMCs with the NPs solvent and polymers solvent (% of control) (Mean ± SEM, n = 4).