Abstract
Bacterial resistance has led to the spread of bacterial infections such as chronic wound infections. Finding solutions for combating resistant bacteria in chronic wounds such as Staphylococcus aureus and Pseudomonas aeruginosa became an attractive theme among researchers. P. aeruginosa is a gram negative opportunistic human pathogenic bacterium that is difficult to treat due to its high resistance to antibiotics. S. aureus (gram negative bacterium) also has a high antibiotic resistance, so that it is resistant to vancomycin (VRSA), tetracycline, fluoroquinolones and beta-lactam antibiotics including penicillin and methicillin (MRSA). In particular, S. aureus and P. aeruginosa have intrinsic and acquired antibiotic resistance, making the clinical management of infection a real challenge, especially in patients with comorbidities. aPDT can be proposed as a new method in the treatment of multi-drug resistant bacteria in chronic wound infection conditions. In this study, the effect of saponin (100 μg/mL) on photodynamic inactivation on planktonic and biofilm forms of P. aeruginosa (ATCC 27853) and S. aureus (ATCC 25923) strains and on Human Dermal Fibroblast (HDF) cells was investigated. Methylene blue (MB) was used as photosensitizer (0, 10, 50, 100 μg/mL). The light source was a red LED source (660 nm; power density: 20 mW/cm(2)) which is related to the maximum absorption of MB. The results showed that the use of saponin in combination with MB-aPDT (Methylene Blue-antibacterial photodynamic therapy) reduces the phototoxic activity of MB due to decreasing the monomer form of MB. This result was obtained by spectrophotometric study. Also, the result of MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide) assay showed that 8 min of irradiation (660 nm) at 10 μg/mL concentration of alone MB had the lowest phototoxic effect on HDF cells. Due to reduced phototoxic properties of MB in this method, detergents containing saponins not recommended to applied at the same time with MB-aPDT in wound infection area.