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Detachment and Killing of Aggregatibacter actinomycetemcomitans Biofilms by Dispersin B and SDS

Department of Oral Biology, New Jersey Dental School, Medical Science Building, Room C-636, 185 S. Orange Ave., Newark, NJ 07103, USA

View larger version (73K): [in this window] [in a new window]   Figure 1. Biofilm growth and detachment of A. actinomycetemcomitans strains CU1000 (wild-type) and HW1018 (PGA mutant) in polystyrene tubes and 96-well microtiter plates. All tubes and microplate wells were stained with crystal violet. (A,B) Biofilm formation at 0 hrs and 24 hrs in tubes (panel A) and microplates (panel B). The biofilms on the right were rinsed with water and treated with SDS (0.1% in PBS) or dispersin B (DspB; 20 µg/mL in PBS) for 5 min prior to crystal violet staining. (C) Detachment of CU1000 biofilms from microplates by SDS. Wells on the bottom were pre-treated with dispersin B for 30 min prior to the SDS treatment.

View larger version (15K): [in this window] [in a new window]   Figure 2. Detachment of CU1000 (wild-type) biofilms from 96-well microtiter plates by SDS. Biofilms were pre-treated with PBS (mock pre-treatment) or dispersin B (20 µg/mL in PBS) for 30 min, and then treated with increasing concentrations of SDS for 5 min. Biofilms were then rinsed and stained with crystal violet. We quantitated the amount of bound crystal violet dye, which is proportional to biofilm biomass, by measuring its absorbance at 590 nm. Values indicate the mean absorbance for duplicate wells. Error bars indicate range.

View larger version (26K): [in this window] [in a new window]   Figure 3. Pre-treatment of A. actinomycetemcomitans CU1000 (wild-type) biofilms with dispersin B increased their sensitivity to killing by SDS. Biofilms grown in polystyrene tubes were rinsed with PBS and treated with 1 mL of PBS (mock pre-treatment) or dispersin B (20 µg/mL in PBS) for 5 min (black bars) or 30 min (gray bars), and then treated with PBS (-) or SDS (0.01% in PBS; +) for 5 min. CFUs were enumerated by dilution plating. Values indicate the log10 of the mean number of CFUs per tube for duplicate tubes. Error bars indicate range.

Journal of Dental Research, Vol. 86, No. 7, 618-622 (2007)
DOI: 10.1177/154405910708600707


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