A new preclinical approach for treating chronic osteomyelitis induced by Staphylococcus aureus: in vitro and in vivo study on photodynamic antimicrobial therapy (PAmT).

Osteomyelitis is an acute or chronic inflammation in the marrow spaces in the superficial or cortical bone, and can be associated with bacterial or fungal infections. Chronic osteomyelitis represents a major health problem due to its difficult treatment and increased morbidity. Photodynamic antimicrobial therapy (PAmT) is a treatment based on a cytotoxic photochemical reaction in which a bright light produced by a laser system and an active photosensitizer absorbed by cells leads to a process of activation that induces a series of metabolic reactions that culminates a bacterial killing. The aim of the present randomized study was to evaluate, by in vitro and in vivo microbiological analysis, the effects of PAmT on tibial surgical bone defects in rats infected by Staphylococcus aureus using bacterial counts carried out immediately and after 30 days after treatment as outcome measure. In the preliminary in vitro study, a diode laser (λ660 nm; 40 mW; ϕ = 0.4 cm(2); 5 or 10 J/cm(2)) and 5, 10, and 15 µg/mL toluidine blue were tested, and the best parameter was chosen for the in vivo study. The concentration of 5 µg/mL was selected to perform the decontamination of S. aureus-infected tibial bone defects in rats. The findings were subjected to statistical analysis. For all PAmTs groups, with the different concentrations, treatment showed significant reductions (p < 0.001) in the amount of bacteria. The in vivo study PAmT group presented a bacterial reduction of 97.4% (p < 0.001). The PAmT using toluidine blue was effective in reducing the number of S. aureus in both in vitro and in vivo studies.

Photodynamic Antimicrobial Chemotherapy (PACT) in osteomyelitis induced by Staphylococcus aureus: Microbiological and histological study.

Osteomyelitis is an inflammation either of medullar spaces or of the surface of cortical bones, which represents a bacterial infection. Photodynamic Antimicrobial Chemotherapy (PACT) is a treatment based on a cytotoxic photochemical reaction that induces a series of metabolic reactions and culminates in bacterial suppression. Such effect led to the idea that it could be used as a treatment of osteomyelitis. Following approval by the Animal Experimentation Committee of the School of Dentistry of the Federal University of Bahia, the present randomized study used eighty Wistar rats with the aim to evaluate, by microbiological and histological analysis, the effects of Photodynamic Antimicrobial Chemotherapy - PACT on tibial surgical bone defects in rats infected by Staphylococcus aureus. The animals were divided in groups: Control (non-infected); Control Osteomyelitis Induction; Saline solution; Photosensitizer; Red Laser and PACT - on this group, a diode laser (40mW; λ660nm ∅=0.04cm(2), CW, 10J/cm(2)) was used in combination with 5µg/ml of toluidine blue as the photosensitizer. On the microbiological study, immediately after treatment, the PACT group presented a bacterial reduction of 97.4% (p<0.001). Thirty days after treatment, there was a bacterial reduction of more than 99.9% (p<0.001). In the histological study, it was observed that the PACT group demonstrated an intense presence of osteocytes and absence of bone sequestration and micro-abscesses. The PACT using toluidine blue was effective in reducing the number of S. aureus enabling a better quality bone repair.