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. 2023 Feb 7;11(2):417. doi: 10.3390/microorganisms11020417

Figure 3.

Figure 3

Ex vivo acne sebaceous gland model with virulent phylotype of C. acnes and S. epidermidis after 48 h of serum SkinDuoTM treatment. (A) Bacterial association with human sebocytes shows result per strain (black bar: C. acnes, grey bar: S. epidermidis). Bacterial enumeration as CFU/mL and expressed as percentage compared to bacteria only showed a decrease in CFU/mL after serum SkinDuoTM treatment, * p < 0.05. (Bi) Conditions tested in 2D human sebocytes for the following: untreated (basal), serum (SkinDuoTM), linoleic acid (positive control for lipid production), TOFA (5-(tetradecyloxy)-2-furoic acid), negative control that reduces fatty acids synthesis and induces inhibition, LPS (lipopolysaccharide) induces an inflammatory challenge; serum in the presence of bacteria (C. acnes, S. epidermidis) and challenged with LPS. The in vitro cellular model challenged with LPS in the presence of bacteria and treated with serum SkinDuoTM significantly reduced total lipid production when compared to bacteria and challenged with LPS. At baseline, the in vitro cellular model treated with serum decreased lipid production. Fluorescence intensity measurement as a ratio of total lipids/nuclei. (Bii) Lipid production analysis was measured with a fluorometer (TECAN) in each well when stained with Hoechst for nuclei ‘DAPI’, Nile Red for neutral lipids ‘FITC’ and total lipids ‘TRITC’, presented as merged picture, lipid droplets are in yellow, and the scale bar represents 20 µm. (C) The evolution rate of metabolic activity over time fluorescence intensity measured with Alamar Blue (Day 4–Day 0/96 h). The reduced fluorescence indicates the inhibition of cellular activity in the presence of bacteria and LPS, the metabolic activity was increased after serum treatment. Evolution rate of metabolic activity, unpaired t-test untreated (sebocytes) Vs: serum, linoleic acid, TOFA, LPS, bacteria only, bacteria + LPS, bacteria + LPS + serum. Each condition was tested in six technical replicates. Error bars represent ±SD. Unpaired t-test: 0.1234 (NS), 0.0332 (*), 0.0002 (***), <0.0001 (****).