Biofilm was grown on stainless steel beads (6mm) using Staphylococcus aureus U1 (Lux UAMS-1; MSSA; generously provided by Dr. Mark Smeltzer) or Pseudomonas aeruginosa (Lux-PAO1, ATCC, Manassas, VA). An isolated colony was inoculated into 3 ml of Mueller–Hinton broth (MHII; Fisher Scientific), and incubated overnight at 37 °C, at 220 RPM. A bacterial suspension of 1 × 106 CFU/ml was created from this culture. Biofilm was produced by placing a 6mm stainless steel bead in a conical tube containing 4 ml of the bacterial suspension and incubated in a shaking incubator at 110 RPM for 2 h at 37 °C. The initial log10CFU bacterial count on the un-implanted balls for S. aureus and P. aeruginosa were on average 5.0 and 4.6, respectively.
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