Fig 1.
Cell-free in vitro H2O2/LPO/SCN- assay mimicking the antimicrobial oxidative response of the Human Airway Epithelium (HAE).
This diagram summarizes how the in vitro test mimics the oxidative response of the HAE.
Table 1.
Microbiological characteristics of clinical strains used in the experiment.
Fig 2.
Kinetic of H2O2 and OSCN- production within the in vitro cell-free assays.
The curves of H2O2 (A) during the H2O2 production assay and OSCN- (B) during the OSCN- production assay, obtained after dosage with ROS-Glo™ H2O2 assay and TNB assay, n = 3 (concentrations calculated for the minimum (green) and maximum (red) height in the well). All relevant data and statistics description are within the supporting information and excel files.
Fig 3.
Kinetic of OSCN- production within the in vitro cell-free assay upon bacteria exposure.
Curves of OSCN- obtained after dosage with TNB. PA14 inoculum of (A) 107 CFU/mL or (B) 102 CFU/mL after 10 hpp (n = 3). PA14 inoculum of (C) 107 CFU/mL or (D) 102 CFU/mL after 15 hpp (n = 3). Concentrations were calculated for the minimum (green) and maximum (red) height in the well. All relevant data and statistics description are within the supporting information and excel files.
Fig 4.
Survival rates of PA14 within the H2O2/LPO/SCN- in vitro assays.
PA14 was incubated in the presence of (A) H2O2 or (B) OSCN- for 1h, 2h and 6h. The bacterial survival was assessed by CFU counting. Mean +/- CI95, n = 10. Data were analysed using a 2-way ANOVA with an P value < 0.05 for inocula as a source of variation in the overall ANOVA. *P <0,05 using Dunnett’s multiple comparisons test. All relevant data are and statistics description within the supporting information and excel files.
Fig 5.
The survival rate of reference and susceptible clinical P. aeruginosa strains within the H2O2 and the H2O2/LPO/SCN- in vitro assays.
The bacterial strains were incubated in the presence of (A and B) H2O2 or (C and D) OSCN- for 1h, 2h and 6h. The bacterial survival was assessed by CFU counting. Mean +/- CI95, n = 10. Data were analysed using a 2-way ANOVA with an P value < 0.05 for inocula as a source of variation in the overall ANOVA. *P <0,05 using Dunnett’s multiple comparisons test. All relevant data and statistics description are within the supporting information and excel files.
Fig 6.
Survival rate of resistant clinical P. aeruginosa strains within the H2O2 and the H2O2/LPO/SCN- in vitro assays.
The bacterial strains were incubated in the presence of (A to D) H2O2 or (E to H) OSCN- for 1h, 2h and 6h. The bacterial survival was assessed by CFU counting. Mean +/- CI95, n = 10. All relevant data and statistics description are within the supporting information and excel files.
Fig 7.
Survival rates of reference and clinical methicillin-susceptible S. aureus strains within the H2O2 and H2O2/LPO/SCN- in vitro assays.
The bacterial strains were incubated in the presence of (A to D) H2O2 or (E to H) OSCN- for 1h, 2h and 6h. The bacterial survival was assessed by CFU counting. Mean +/- CI95, n = 10. MSSA: Methicillin-susceptible S. aureus. All relevant data and statistics description are within the supporting information and excel files.
Fig 8.
Survival rates of reference and clinical methicillin-resistant S. aureus strains within the H2O2 and H2O2/LPO/SCN- in vitro assays.
The bacterial strains were incubated in the presence of (A to C) H2O2 or (D to F) OSCN- for 1h, 2h and 6h. The bacterial survival was assessed by CFU counting. Mean +/- CI95, n = 10. MRSA: Methicillin-resistant S. aureus. All relevant data and statistics description are within the supporting information and excel files.
Fig 9.
Schematic of the H2O2/LPO/SCN- in vitro assays developed in our article.
This diagram summarizes the analytic workflow for our in vitro test. Created in BioRender.com.
Fig 10.
ROS action on P. aeruginosa and S. aureus.
Schematic for the potential mechanisms explaining the higher sensitivity to ROS of P. aeruginosa vs S. aureus and P. aeruginosa. Created in BioRender.com.