Journal: bioRxiv

Article Title: An ex vivo human model for safety assessment of immunotoxicity of engineered nanomaterials

doi: 10.1101/2023.06.29.547008

Figure Lengend Snippet: (A) by silica ENM with different surface functionalities. PBMCs response to Silica ENM (25 nm diameter) exposure (4 h treatment) was assessed in the naive state (2 h pre-incubation with medium) and in the primed state (2 h pre-incubation with LPS). Propidium iodide (PI, 5 µg/ml) was added for the last 30 min of the incubation time prior to fixation, after which cells were stained for inflammasome markers and analyzed by high-content imaging. Measurements (diamonds for the naive state and dots for the primed state) represent the mean of 16 image fields acquired. The lines show the result of a four-parametric logistic regression to determine EC 50 values. The black dotted line shows the median + 3std as the activity threshold of non-ENM treated (B) naive control for PI-positive cells (17%) and (C) primed control for inflammasome activationrate (41%), n=48 wells with 16 images per well. (B) PI-positive cells are expressed as percentages relative to the total cell count. (C) Activationrate calculated as a percentage relative to CD14 + Monocytes.

Article Snippet: 5 min prior to compound addition, samples are placed on an orbital shaker for 5 min. After 2 h of incubation with LPS, ENM and solvent controls (in different doses) were added and incubation was continued for more than 3.5 hours when 10 µl of Hoechst/propidium iodide (H33342: 2 µg/ml, PI: 5 µg/ml, Sigma-Aldrich) staining and ATP (2 mM final concentration, Sigma-Aldrich) activator control was added.

Techniques: Incubation, Staining, Imaging, Activity Assay, Cell Counting

Journal: International Journal of Nanomedicine

Article Title: Platelet-functionalized three-dimensional poly-ε-caprolactone fibrous scaffold prepared using centrifugal spinning for delivery of growth factors

doi: 10.2147/IJN.S120206

Figure Lengend Snippet: Visualization of MG-63 cells on scaffolds prepared using centrifugal spinning. Notes: Cells were stained using DiOC6 (internal cell membranes, green color) and propidium iodide (cell nuclei, red color) and visualized using a confocal microscope. Cells adhered similarly on all the tested scaffolds. The highest number of cells was visible on the PCL/P1 on day 7. On day 14, the cell number was decreasing. The numbers of cells on the scaffolds with low concentrations of platelets (PCL/P3, PCL/P4) and control sample (PCL) were constant during the experiment. The results correspond well with the data from DNA analysis. Abbreviations: DiOC6, 3,3′-diethyloxacarbocyanine iodide; PCL, poly-ε-caprolactone; P, platelets.

Article Snippet: Subsequently, the samples were rinsed with PBS, and propidium iodide (PI; 5 µg/mL in PBS; Sigma-Aldrich) was added for 10 min, followed by rinsing with PBS.

Techniques: Staining, Microscopy