The Effect of 532 nm Diode Pumped Solid State (DPSS) Laser in Combination with Safranin on the Growth of Pseudomonas aeruginosa and Staphylococcus aureus in Vitro

Abstract

The effect of 532nm Diode Pumped Solid State (DPSS) laser at power density of 5.234 W/cm2 on the growth of Gram-negative Pseudomonas aeruginosa and Gram-positive Staphylococcus aureus was evaluated. These bacteria were isolated from samples taken from burn and infected wound areas of 55 patients admitted to the burn-wound unit in Al-Kindy teaching hospital in Baghdad during the period from October 2012 to March 2013. Each isolate was identified using microscopic, cultural and biochemical methods. A standard bacterial suspension was prepared for each isolate. Serial dilutions were then prepared and a dilution of 10-5 was selected. Irradiation experiments included four groups: (L-P-) bacterial suspension in saline solution, (L-P+) bacterial suspension in the presence of 0.1mg/ml photosensitizer (safranin O), (L+P-) bacterial suspension treated with laser radiation only and finally (L+P+) bacterial suspension treated with laser radiation in the presence of the photosensitizer. After irradiation, Cetrimide agar (P. aeruginosa) and Mannitol salt agar (S.aureus) were used. Seven replicates were used for each experimental group. Different times of exposure were applied for irradiated groups. The results revealed that twenty two isolates out of 70 samples were positive for P. aeruginosa (31%) and fifteen isolates out of 35 samples were positive for S. aureus (42%). The combined effect of DPSS laser and safranine O was significantly effective in reducing the number of Colony Forming Units per milliliter (CFU/ml) of P. aeruginosa and S. aureus compared with control groups. Almost a complete bacterial mortality achieved at 25 and 5 minutes of exposing to laser light in the presence of safranin O for P. aeruginosa and S. aureus respectively, suggesting that singlet oxygen (O2*) and/or reactive oxygen species (ROS) were involved in the killing of the bacteria.