Biofilm-related infections are serious problems in the Orthopedics field, and Staphylococcus aureus are the most popular causative agents of bacterial infections associated with arthroplasty. Several studies demonstrated a synergistic effect of the electric intervention (EI) and the antibiotic administration in killing bacteria in biofilm; however, a constant, long-time EI was needed. In the present study, the effective removal of biofilm formed with S. aureus on a titanium ring by multiple times of one minute-EI was observed and described.
A methicillin-sensitive S. aureus clinical isolate was used to form biofilm on a titanium ring. After applying a series of EI with various combinations of the frequencies and timings, the amount and principal components of biofilms were assessed with crystal violet staining, live bacterial cell count, and fluorescence staining with confocal laser scanning microscopy.
More than 60% biofilm removal was observed in the 2-time EI applied at 24 (1) and 72 (3) h (days) post bacterial exposure (PBE) and in the 3-time EI at 0 (0), 24 (1), and 72 (3) h (days) PBE, or at 24 (1), 48 (2), and 72 (3) h (days) PBE. The live bacterial cell numbers, the proportion of live and dead cells, and the amount of extracellular polysaccharide substances (EPS) of biofilm were similar with or without EI. It was assumed that an excess amount of the biofilm removal shown in the several EI was not attributed to the effect of the electrolysis.
The effective removal of biofilm was observed when multiple times 1 min EI was applied without any changes in the proportion of live and dead bacteria or the amount of EPS. The mechanisms to explain extra biofilm removal remain to be elucidated.

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