Fig 1.
Log10 CFU/mL for both C. albicans and S. aureus strains in single- and dual-species biofilms.
Individual statistical analyses were conducted only for comparison between the negative control group, ACP group and positive control group (i.e. penicillin for S. aureus and fluconazole for C. albicans) of the same strain in each biofilm. * different letters denote statistically significant difference (p<0.05) only between groups of the same strain in each graph.
Fig 2.
Confocal laser scanning microscopy of C. albicans biofilm in negative control group, ACP group and positive control group (fluconazole); 4 × zoom.
Biofilm stained with Live/Dead BacLight Viability kit. Live cells in fluorescent green and dead cells in fluorescent red. 2D view (left side image), surface plot of 3D volume image (center image), and cross-section of 3D volume image (right side image) show the distribution of live and dead cells throughout biofilm layers.
Fig 3.
Confocal laser scanning microscopy of S. aureus biofilm in negative control group, ACP group and positive control group (penicillin); 4 × zoom.
Biofilm stained with Live/Dead BacLight Viability kit. Live cells in fluorescent green and dead cells in fluorescent red. 2D view (left side image), surface plot of 3D volume image (center image), and cross-section of 3D volume image (right side image) show the distribution of live and dead cells throughout biofilm layers.
Fig 4.
Confocal laser scanning microscopy of C. albicans + S. aureus biofilm in negative control group, ACP group and positive control groups (fluconazole and penicillin); 4 × zoom.
Biofilm stained with Live/Dead BacLight Viability kit. Live cells in fluorescent green and dead cells in fluorescent red. 2D view (left side image), surface plot of 3D volume image (center image), and cross-section of 3D volume image (right side image) show the distribution of live and dead cells throughout biofilm layers.
Fig 5.
SEM micrographs of single and dual-species biofilms in negative control group, ACP group and positive control groups (fluconazole and penicillin); 5000 × zoom.
C. albicans (CA), S. aureus (SA) and C. albicans + S. aureus (CA/SA) biofilms. Note rupture of cells architecture in all biofilms after ACP treatment (groups CA-ACP, SA-ACP and CA/SA-ACP).
Fig 6.
Fluorescence microscopy images of single and dual-species biofilms in negative control group and ACP group; 1.6 × lens and 5.0 × zoom.
Fluorescent green (oxidized rhodamine 123) represents presence of intracellular ROS. CA—C. albicans biofilm, SA—S. aureus biofilm, and CA/SA—C. albicans + S. aureus biofilm.
Fig 7.
Mean (± SD) percent cytotoxicity values of ROE for negative control group, positive control group and ACP group.
Fig 8.
Mean (± SD) percent viability values of ROE for negative control group, positive control group and ACP group.
Fig 9.
Oral epithelium sections of negative control group, positive control group and ACP group; 20 × zoom.
Sections stained by hematoxylin and eosin for histology (top) and immunohistochemistry for Ki67 (bottom).