Journal: Materials Today Bio

Article Title: Viridibacillus culture derived silver nanoparticles exert potent anticancer action in 2D and 3D models of lung cancer via mitochondrial depolarization-mediated apoptosis

doi: 10.1016/j.mtbio.2024.100997

Figure Lengend Snippet: Study of migration of 2D cultured A549 cells in absence and presence of epidermal growth factor (EGF) and V-AgNPs at LD50-2D. (A) The schematic shows creation of cell-free area using cell inserts and addition of cell culture medium (control group), EGF (positive control group), and EGF + V-AgNPs (treatment group). Upon these treatments, the time lapse imaging was done to understand A549 cell migration in cell-free area using IncuCyte S3 live cell imaging platform. (B) The schematic shows the image analysis pipeline for time lapse images obtained from IncuCyte S3 instrument. The image analysis was done using our previously developed macro script for understanding % cell-free are over time as well as using a Matlab-GUI based CellTracker program for understanding cell migration pattern and migration velocities under various treatments. (C) The binary time lapse images obtained after image analysis show percent reduction of cell-free area (white area in the images) due to migration of A549 cells (black area in the images) in presence of only cell culture medium (black, upper row), EGF (blue, middle row), and EGF + V-AgNPs (red, lower row). The % cell-free areas were calculated by considering the cell-free area of 0 th h images as 100%. (D) The line graph shows the cell migration kinetics under the influence of various treatments, where each line represents mean ± std. error of 3 replicates ( n = 3). The yellow box indicates the time frame within which significant differences (two-way ANOVA, p-value <0.05) were observed in each treatment. The bar graph showing these differences is given in supporting information, . (E) The bar graph represents average cell migration velocities obtained from CellTracker program and each bar represents mean velocity ± std. error of randomly selected cells ( n = 10). The cell migration patterns and directionalities in presence of aforementioned treatments are given in the supporting information, . The asterisks in the bar graph represent significantly different observations (One-way ANOVA test: **** p-value<0.0001). (Abbreviations – V-AgNPs: Viridibacilli derived silver nanoparticles, EGF: Epidermal growth factor, t 0 : initial time or start time, t’: time point other that t 0 , h: hours, ns: not significant) (Schematics were created with BioRender.com ). (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: The image analysis was done using our previously developed macro script for understanding % cell-free are over time as well as using a Matlab-GUI based CellTracker program for understanding cell migration pattern and migration velocities under various treatments. (C) The binary time lapse images obtained after image analysis show percent reduction of cell-free area (white area in the images) due to migration of A549 cells (black area in the images) in presence of only cell culture medium (black, upper row), EGF (blue, middle row), and EGF + V-AgNPs (red, lower row).

Techniques: Migration, Cell Culture, Control, Positive Control, Imaging, Live Cell Imaging, Derivative Assay