title: Effectiveness of antimicrobial photodynamic therapy using a 660 nm laser and methyline blue dye for inactivating Staphylococcus aureus biofilms in compact and cancellous bones: An in vitro study.
abstract: BACKGROUND
  New therapeutic modalities such as antimicrobial photodynamic therapy (APDT) has been investigated in order to be a valid alternative to the treatment of infections caused by different microorganisms. This work evaluated the in vitro effectiveness of Antimicrobial Photodynamic Therapy (APDT) using 660 nm laser combined with methylene blue dye to inactivate Staphylococcus aureus (ATCC 25923) biofilms in compact and cancellous bones specimens.
  
  
  METHODS
  Eighty specimens of compact bone and 80 specimens of cancellous bone were contaminated with a standard suspension of S. aureus and incubated for 14 days at 37°C to induce the formation of biofilms. The specimens were then divided into groups (n = 10) according to the established treatment: PS-L- (control--no treatment), PS+L- (only AM for 5 min in the dark), PS-L+90 (only laser irradiation for 90 s), PS-L+180 (only laser irradiation for 180 s), PS-L+300 (only laser irradiation for 300 s), APDT90 (APDT for 90 s), APDT180 (APDT for 180 s), and APDT300 (APDT for 300 s). The findings were statistically analyzed by ANOVA 5%.
  
  
  RESULTS
  All of the experimental treatments showed a significant reduction (log 10 CFU/mL) of S. aureus biofilms in compact and cancellous bones specimens compared with the control group, and the APDT group was the most effective. Compact specimens treated with APDT showed the greatest reduction in biofilms compared with cancellous specimens, regardless of length of treatment.
  
  
  CONCLUSIONS
  APDT with methylene blue dye and a 660 nm laser proved to be effective in inactivating S. aureus biofilms formed in compact and cancellous bone.
authors.affiliation: Multidisciplinary Health Institute, Federal University of Bahia, Biointegration, Rua Rio de Contas, 58 Vitória da Conquista, BA, Brazil. Electronic address: drlucianorosa@yahoo.com.br.
authors.forename: Luciano Pereira
authors.initials: LP
authors.lastname: Rosa
authors.affiliation: Multidisciplinary Health Institute, Federal University of Bahia, Biointegration, Rua Rio de Contas, 58 Vitória da Conquista, BA, Brazil. Electronic address: drfransilva@yahoo.com.br.
authors.forename: Francine Cristina da
authors.initials: FC
authors.lastname: Silva
authors.affiliation: Multidisciplinary Health Institute, Federal University of Bahia, Biointegration, Rua Rio de Contas, 58 Vitória da Conquista, BA, Brazil. Electronic address: sumaianader@yahoo.com.br.
authors.forename: Sumaia Alves
authors.initials: SA
authors.lastname: Nader
authors.affiliation: Multidisciplinary Health Institute, Federal University of Bahia, Biointegration, Rua Rio de Contas, 58 Vitória da Conquista, BA, Brazil. Electronic address: gih.andradem@gmail.com.
authors.forename: Giselle Andrade
authors.initials: GA
authors.lastname: Meira
authors.affiliation: Multidisciplinary Health Institute, Federal University of Bahia, Biointegration, Rua Rio de Contas, 58 Vitória da Conquista, BA, Brazil. Electronic address: magda---viana@hotmail.com.
authors.forename: Magda Souza
authors.initials: MS
authors.lastname: Viana
mesh_terms: D000818:Animals; D000890:Anti-Infective Agents / Q000627:therapeutic use*; D018441:Biofilms / Q000187:drug effects*; D001842:Bone and Bones / Q000187:drug effects / Q000382:microbiology; D002417:Cattle; D007834:Lasers; D008751:Methylene Blue; D010778:Photochemotherapy / Q000379:methods*; D017319:Photosensitizing Agents / Q000627:therapeutic use*; D013211:Staphylococcus aureus / Q000187:drug effects*
chemical_list: D000890:Anti-Infective Agents; D017319:Photosensitizing Agents; D008751:Methylene Blue
keywords: Biofilm; Methylene blue; Microorganism; Photodynamic therapy; Staphylococcus aureus