93 J/cm2, with stirring. Three additional wells containing 50 μL of methylene blue and
50 μL of the bacterial suspension were kept in the dark to assess the toxicity of the photosensitiser alone. To assess the toxicity of laser light alone, ACY-1215 nmr 50 μL PBS was added to 50 μL of the inoculum in a further six wells, three of which were irradiated with laser light and the remaining three kept in the dark. Following irradiation/dark incubation, samples were serially diluted 10-fold in PBS and plated onto 5% horse blood agar plates in triplicate. The plates were incubated aerobically overnight at 37°C, following which the surviving CFU/mL were enumerated by viable counting. Experiments were performed three times in triplicate. To examine the effect of laser light dose on the photodynamic killing of the SCVs, methylene blue was diluted in PBS to give a final concentration of 20 μM. Experiments were performed as described above, but bacteria were irradiated with 1.93 J/cm2, 3.86 J/cm2 or 9.65 J/cm2 of 665 nm laser light, with stirring. Following irradiation/dark incubation, viable bacteria SAHA HDAC were enumerated as described as above. Acknowledgments John Wright and Sean Nair received funding from the charity Arthritis Research UK (grant number 18294).
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