Combining Visible Light and Non-Focused Ultrasound Significantly Reduces <em>Propionibacterium acnes</em> Biofilm While Having Limited Effect on Host Cells

Abstract: Bacterial biofilms are highly resistant to antibiotics and have been implicated in the etiology of 60-80% of chronic microbial infections. We tested a novel combination of low intensity ultrasound and blue light against biofilm and planktonic bacteria. A laboratory prototype was built which produced both energies uniformly and coincidently from a single treatment head, impinging upon a 4.45 cm^2 target. To demonstrate proof of concept, Propionibacterium acnes biofilm was cultured on Millicell hanging inserts in 6-well plates. Hanging inserts with biofilm were treated in a custom exposure chamber designed to minimize unwanted ultrasound reflections. Coincident delivery of both energies demonstrated synergy over either alone, killing both stationary planktonic and biofilm cultures of P. acnes. Biofilm killing was dose dependent on exposure time (i.e. energy delivered). P. acnes biofilms were significantly reduced by the dual energy treatment (p<0.0001), with a >1 log10 reduction after a 5 min (9 J/cm^2) and >3 log10 reduction after a 30 min (54 J/cm^2) treatment (p<0.05). Mammalian cells were found unaffected by the treatment. Both the light and the ultrasound energies are at levels previously cleared by the FDA. Therefore, this combination treatment could be used as a safe, efficacious method to treat biofilm related syndromes.