Journal: Nutrients

Article Title: Carvacrol Suppresses Inflammatory Biomarkers Production by Lipoteichoic Acid- and Peptidoglycan-Stimulated Human Tonsil Epithelial Cells

doi: 10.3390/nu14030503

Figure Lengend Snippet: The effect of carvacrol on the viability of human tonsil epithelium cells; ( A ) Cell viability changes with 16.1, 31.5, 62.5, 125, and 250 µg/mL of carvacrol in DMSO, 4, 8, and 16.1 µg/mL of nimesulide, lipoteichoic acid (LTA) + peptidoglycan (PGN) (10 µg/mL each), and 0.05% dimethyl sulfoxide (DMSO) for 24 h were determined using 7-AAD staining followed by flow cytometry (FCM) analysis of human tonsil epithelial cells (HTonEpiCs). Absorbance was measured at 488 nm; Cell viability (%) was calculated relative to the control of 0.05% DMSO; Values are shown as mean ± SE from three independent experiments, each in triplicate; *, The different letters above the columns show that the means of different groups were significantly different ( p < 0.05) by one-way analysis of variance using Tukey’s test; ( B ) The overlay of histograms and scatter plots of the controls and samples given by FCM analysis. ( C ) The morphological changes of the HTonEpiCs cells after the treatments were examined under an inverted microscope at 10 × 40 magnification; Representative photographs that were taken 24 h after the treatment in three independent experiments are presented; ( a ) untreated; ( b ) vehicle control (0.25% DMSO); and ( c ) 16 µg/mL carvacrol; ( d ) 31.5 µg/mL carvacrol; ( e ) 62.5 µg/mL carvacrol; ( f ) 125 µg/mL carvacrol; ( g ) 250 µg/mL carvacrol; ( h ) LTA + PGN (10 µg/mL, each); and ( i ) 8 µg/mL nimesulide. Scale bar = 1 mm. 7-AAD: 7-amino-actinomycin D; DMSO: dimethyl sulfoxide; LTA: lipoteichoic acid; and PGN: peptidoglycan.

Article Snippet: Effects of carvacrol on cell viability (proliferation) were analyzed by flow cytometry using an AttuneTM NxT acoustic focusing flow cytometer (AFC2, Thermo Fisher Scientific Inc., San Jose, CA, USA) and AttuneTM NxT flow cytometry analysis software (v3.1.1234.0., Thermo-Fisher Scientific Inc., San Jose, CA, USA).

Techniques: Staining, Flow Cytometry, Control, Inverted Microscopy