Endang W. Bachtiar, Sari Dewiyani, Siti M. Surono Akbar, Boy M

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Inhibition of Candida albicans biofilm development by unencapsulated Enterococcus faecalis cps2 Endang W. Bachtiar, Sari Dewiyani, Siti M. Surono Akbar, Boy M. Bachtiar Journal of Dental Sciences Volume 11, Issue 3, Pages 323-330 (September 2016) DOI: 10.1016/j.jds.2016.03.012 Copyright © 2016 Terms and Conditions

Figure 1 Stain-all staining of heat extracts of different Enterococcus faecalis strains after electrophoresis in an acrylamide gel. Lanes: 1 = prestained protein markers (New England BioLabs); 2 = reference strain (ATCC 29212); 3 = encapsulated E. faecalis cps2 (strain that showed no positive result after qPCR); 4 = unencapsulated cps2 (strain that showed positive result after qPCR). The arrow indicates locations of CPS. CPS = capsular polysaccharide; qPCR = quantitative polymerase chain reaction. Journal of Dental Sciences 2016 11, 323-330DOI: (10.1016/j.jds.2016.03.012) Copyright © 2016 Terms and Conditions

Figure 2 Effects of unencapsulated Enterococcus faecalis cps2 or E. faecalis reference strain (ATCC 29212) or their respective spent medium, which contains 10 μg/mL protein, on established Candida albicans ATCC 10231 biofilms and cell viability within the biofilms. After each time point (24 hours and 48 hours). (A) Biofilm masses were quantified using crystal violet staining and (B) viability cell within the remaining biofilms were assessed using CFU evaluation. Data are means ± standard deviations of three independent experiments performed in triplicate. * Statistical significance (P < 0.05) of observed differences between each time point. CFU = colony-forming unit; OD = optical density; SM-ATCC 29212 = spent medium of the reference strain; SM-UEs = spent medium of unencapsulated E. faecalis strain; UEs = unencapsulated E. faecalis strain. Journal of Dental Sciences 2016 11, 323-330DOI: (10.1016/j.jds.2016.03.012) Copyright © 2016 Terms and Conditions

Figure 3 ALS1, ALS3, and EFB1 gene expression levels at different time points (24 hours and 48 hours) during Candida albicans biofilm development in the presence of unencapsulated Enterococcus faecalis cps2 or E. faecalis ATCC 29212. Data are shown as the fold regulation calculated using the comparative Ct method. The tested genes were normalized to the ACT1 gene level. * Indicates a statistically significant difference (P < 0.05) in reducing EFB1 gene between the E. faecalis strains tested after 48 hours. UEs = unencapsulated E. faecalis strain. Journal of Dental Sciences 2016 11, 323-330DOI: (10.1016/j.jds.2016.03.012) Copyright © 2016 Terms and Conditions

Figure 4 Spent medium, containing 10 μg/mL protein, from unencapsulated Enterococcus faecalis cps2 strain or E. faecalis ATCC 29212 that was added into Candida albicans biofilm for 24 hours and 48 hours, respectively. Means of ALS1, ALS3, and EFB1 transcription level were deduced from three separate experiments is presented with SD. * Indicates statistically significant (P < 0.05) difference between transcription level of the EFB1 genes in C. albicans biofilm, compared with the negative control. SD = standard deviation; UEs = unencapsulated E. faecalis strain. Journal of Dental Sciences 2016 11, 323-330DOI: (10.1016/j.jds.2016.03.012) Copyright © 2016 Terms and Conditions