Clorox ™ ,Peridex ™ & 3 commercial Denture Cleansers as Denture Disinfecting Agents; In Vitro Comparison Study

1. Clorox™,Peridex™ & 3 commercial Denture Cleansers as Denture Disinfecting Agents;In Vitro Comparison Study A.A.DAHLAN* ,G. RAMAGE, C. HAVEMAN, S. REDDING, JL. LOEZ-RIBOT, L.BRADLY.UTHSCSA,TX.

2. Objectives • Determine the lowest concentration of Clorox™ (Sodium hypochlorite)and Peridex ™(Chlorhexidine) that eliminate the biofilm metabolic activity (BMA) of C.albicans. • Determine the effectiveness of solutions of three commercial denture cleansers(Efferdent ™,Polident ™ for Dentures & Polident ™ for Partials) against BMA of C.albicans.

3. Definitions • Biofilm: a matrix-enclosed population of cells adherent to surfaces, intersurfaces,and/or each other in a complex three-dimensional structure. • Denture Induced Stomatitis (Chronic Atrophic Candidiasis): a very common oral fungal infection among denture wearers caused principally by Candida albicans as a result of diminished host resistance. • XTT metabolic assay: spectrophometric assay that measure the metabolic activity of the biofilms.

4. Significance of the study • Disinfect dentures contaminated with microorganisms (usually C.albicans). • Long term goal is to develop a simple, inexpensive & effective protocol to disinfect dental appliances with or without metal components that can be easily accomplished by patients in their home. • The use of XTT metabolic assay.

5. Methods & Material • Organisms: 4 strains of C.albicans were isolated from denture stomatitis patients (3153A,SC5314,6455 & 6309) • Biofilm formation: 100µl of standardized cell suspension(1x10*cell/ml) was pipetted into selected wells of microtiter plates and incubated for 48 hours @37°C then medium was aspirated and non-adherent cells removed by thorough washing with PBS. • Disinfectants: • . Clorox™ added in serially double diluted concentrations up to 1/512th & incubated for 30 min. @ room Temp. • Peridex ™ added at 3 conc.(100%, 50% & 25%) incub. 30 min @ room Temp • Efferdent ™, Polident-D ™ & Polident-P ™ tab. were dissolved in 200 ml of sterile water . 2 trails:incub. 30 min + incub. 18 hr.

6. +ve and –ve controls were included • Biofilms were washed three times in sterile PBS

7. XTT Reduction Assay: • 100µl aliquot of XTT was added to each prewashed biofilm and to control wells to measure the background XTT levels then plates were incubated in the dark for 1 hr @37°C. A colormetric change of XTT, a direct reflection of the metabolic activity of the biofilm, was then measured in a microtiter plate reader at 490nm.

8. Results

9. The Effect of Clorox ™ on C.Albicans biofilm

10. The Effect of Peridex ™ on C.Albicans biofilm

11. The Effect of Polident ™ -D,Polident ™ -P & Efferdent ™ on C.Albicans biofilm”30 min. Tx

12. The Effect of Polident ™ -D,Polident ™ -P & Efferdent ™ on C.Albicans biofilm”18 hr Tx.”

13. Conclusion There was a 100% decrease in the biofilm metabolic activity of all tested strains exposed for 30 min. to Clorox ™ in concentration of 1:32 and stronger. There was also a 100% decrease in the biofilm metabolic actvitiy of all tested strains except SC5314 when treated with Peridex ™ in concentration of 100% & 50% for 30 min or Polident ™ for dentures for 18hr.