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Lasers in Medical Science

, Volume 33, Issue 8, pp 1723–1731 | Cite as

Bactericide effect of methylene blue associated with low-level laser therapy in Escherichia coli bacteria isolated from pressure ulcers

  • Thais Ferreira Gomes
  • Matheus Masalskiene Pedrosa
  • Ana Claudia Laforga de Toledo
  • Veridiana Wanshi Arnoni
  • Mirian dos Santos Monteiro
  • Davi Cury Piai
  • Silvia Helena Zacarias Sylvestre
  • Bruno FerreiraEmail author
Original Article

Abstract

The present study analyzed the bactericidal effect of methylene blue associated with low-level lasers on Escherichia coli isolated from a pressure ulcer. Microbiological material from a pressure ulcer was isolated using an aseptic swab, and antimicrobial activity was verified using the diffusion disc method. Methylene blue was used at concentrations of 0.001 and 0.005%, and low-level lasers of 670, 830, and 904 nm, with the energy densities of 4, 8, 10, and 14 J/cm2, were tested on three plates each and combined with methylene blue of each concentration. In addition, three control plates were used, with each concentration and energy density separated without any interventions. The results were analyzed using the paired sample t test to determine the bactericidal effect of the methylene blue and using the ANOVA test to compare the effects of the energy densities and wavelengths among the low-level laser treatment protocols. The results showed bacterial reduction at wavelengths of 830 and 904 nm and more proliferation in wavelengths of 670 nm. In wavelength of 830 nm, a bacterial reduction was observed in the conditions with 0.001% methylene blue in all energy density utilized, with 0.005% methylene blue in energy density of 10 J/cm2, and without methylene blue in energy density at 10 J/cm2. And in a wavelength of 904 nm, all condition showed bacterial reduction with or without methylene blue. We concluded that the low-level lasers of 904 and 830 nm have bactericidal effects and at better energy densities (10 and 14 J/cm2).

Keywords

Methylene blue Low-level laser Pressure ulcer 

Notes

Acknowledgements

We thank the Department of Physical Therapy and Study Center and Research on Regional Development (CEPed) of the University Center UNIFAFIBE, SP.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

This research was approved by the Research Ethics Committee (number CAAE: 67741017.4.0000.5387).

References

  1. 1.
    Sen CK, Gordillo GM, Roy S et al (2009) Human skin wounds: a major and snowballing threat to public health and the economy. Wound Repair Regen 17:763–771CrossRefGoogle Scholar
  2. 2.
    Kim H, Chung H, Wang S et al (2014) SAPPIRE: a prototype mobile tool for pressure ulcer risk assessment. Stud Health Technol Inform 201:433–440PubMedPubMedCentralGoogle Scholar
  3. 3.
    Walker RM, Gillespie BM, Thalib L et al (2017) Foam dressings for treating pressure ulcers. Cochrane Database Syst Rev 10:113–132Google Scholar
  4. 4.
    David OP, Sierra-Sosa D, Zapirain BG (2017) Pressure ulcer image segmentation technique through synthetic frequencies generation and contrast variation using toroidal geometry. Biomed Eng Online 16:1–19CrossRefGoogle Scholar
  5. 5.
    Braga IA, Brito CS, Filho AD et al (2017) Pressure ulcer as a reservoir of multiresistant Gram-negative bacilli: risk factors for colonization and development of bacteremia. Braz J Infect Dis 21:171–175CrossRefGoogle Scholar
  6. 6.
    Arzt H, Fromanrin I, Ribinik P et al (2012) Which medical device and/or which local treatment are to be used, as of 2012, in patients with infected pressure sore? Developing French guidelines for clinical practice. Ann Phys Rehabil Med 55:498–507CrossRefGoogle Scholar
  7. 7.
    Qaseem A, Humphrey LL, Forciea MA (2015) Treatment of pressure ulcers: a clinical practice guideline from the American College of Physicians. Ann Intern Med 162:370–380CrossRefGoogle Scholar
  8. 8.
    Palagi S, Severo IM, Menegon DB, Lucena AF (2015) Laser therapy in pressure ulcers: evaluation by the Pressure Ulcer Scale for Healing and Nursing Outcomes Classification. Rev Esc Enferm USP 49:820–826CrossRefGoogle Scholar
  9. 9.
    Andrade FSSD, Clark RMO, Ferreira ML (2014) Effects of low-level laser therapy on wound healing. Rev Col Bras Cir 41:129–133CrossRefGoogle Scholar
  10. 10.
    Pandey R, Koppulo P, Kalakonda B et al (2017) Treatment of dentinal hypersensitivity using low-level laser therapy and 5% potassium nitrate: a randomized, controlled, three arm parallel clinical study. Int J Appl Basic Med Res 7:63–66CrossRefGoogle Scholar
  11. 11.
    Ganvir S, Agrawal M, Hashchandre M (2016) Combined effect of ultrasound and laser therapy (LLLT) for the treatment of pressure ulcer in a patient with spinal cord injury. Physiother Rehabil 1:1–3CrossRefGoogle Scholar
  12. 12.
    Kajagar BM, Godhi AS, Pandit A, Khatri S (2012) Efficacy of low level laser therapy on wound healing in patients with chronic diabetic foot ulcers—a randomised control trial. Indian J Surg 74:359–363CrossRefGoogle Scholar
  13. 13.
    Carvalho PTC, Marques APC, Reis FA et al (2006) Photodynamic inactivation of in vitro bacterial cultures from pressure ulcers. Acta Cir Bras 21:32–35CrossRefGoogle Scholar
  14. 14.
    Lambrechts SA, Demidova TN, Aalders MC et al (2005) Photodynamic therapy for Staphylococcus aureus infected burn wounds in mice. Photochem Photobiol Sci 4:503–509CrossRefGoogle Scholar
  15. 15.
    Sebrão CC, Bezerra AG Jr, de França PH et al (2017) Comparison of the efficiency of Rose Bengal and methylene blue as photosensitizers in photodynamic therapy techniques for Enterococcus faecalis inactivation. Photomed Laser Surg 35:18–23CrossRefGoogle Scholar
  16. 16.
    Gardner SE, Frantz RA, Saltzman CL et al (2006) Diagnostic validity of three swab techniques for identifying chronic wound infection. Wound Repair Regen 14:548–557CrossRefGoogle Scholar
  17. 17.
    Zenuz AT, Eslami H, Kafil HS et al (2016) The application of antimicrobial photodynamic therapy on Pseudomonas aeuroginosa and Enterococcus fecalis using hypericin and methylene blue photosensitizers. Biomed Pharmacol J 9:443–450CrossRefGoogle Scholar
  18. 18.
    CLSI (2012) Performance Standards for Antimicrobial Disk Susceptibility Tests; Approved Standard-Eleventh Edition Clinical and Laboratory Standards Institute 32Google Scholar
  19. 19.
    Wong-Leung YL (1988) Antibacterial activities of some Hong Kong plants used in Chinese medicine. Fitoterapia 69:11–16Google Scholar
  20. 20.
    Naovi SA, Khan MS, Vohora SB, Naqvi S (1991) Antibacterial, anti-fungal and anthelmintic investigations on Indian medicinal plants. Rev. Fitoterapia 62:221–228Google Scholar
  21. 21.
    Brugger SD, Baumberger C, Jost M et al (2012) Automated counting of bacterial colony forming units on agar plates. PLoS One 7:1–6Google Scholar
  22. 22.
    Livesley NJ, Chow AW (2002) Infected pressure ulcers in elderly individuals. Clin Infect Dis 35:1390–1396CrossRefGoogle Scholar
  23. 23.
    Kharkwal GB, Sharma SK, Huang YY et al (2011) Photodynamic therapy for infections: clinical applications. Lasers Surg Med 43:755–767CrossRefGoogle Scholar
  24. 24.
    Oliveira BP, Lins CCSA, Diniz FA et al (2014) In vitro antimicrobial photoinactivation with methylene blue in different microorganisms. Braz J Oral Sci 13:53–57CrossRefGoogle Scholar
  25. 25.
    Moura-Neto C, Ferreira LS, Maranduba CM et al (2016) Low-intensity laser phototherapy enhances the proliferation of dental pulp stem cells under nutritional deficiency. Braz Oral Res 30:1–6Google Scholar
  26. 26.
    Asnaashari M, Godiny M, Azari-Marhabi S et al (2016) Comparison of the antibacterial effect of 810 nm diode laser and photodynamic therapy in reducing the microbial Flora of root canal in endodontic retreatment in patients with Periradicular lesions. J Lasers Med Sci 7:99–104CrossRefGoogle Scholar
  27. 27.
    Calderhead RG, Kim Q-S, Ohshiro T et al (2015) Adjunctive 830 nm light-emitting diode therapy can improve the results following aesthetic procedures. Laser Ther 24:277–289CrossRefGoogle Scholar
  28. 28.
    Malgikar S, Reddy SH, Sagar SV et al (2016) Clinical effects of photodynamic and low-level laser therapies as an adjunct to scaling and root planing of chronic periodontitis: a split-mouth randomized controlled clinical trial. Indian J Dent Res 21:121–126CrossRefGoogle Scholar
  29. 29.
    Tennert C, Feldmann K, Haamann E et al (2014) Effect of photodynamic therapy (PDT) on enterococcus faecalis biofilm in experimental primary and secondary endodontic infections. BMC Oral Health 14:1–8CrossRefGoogle Scholar
  30. 30.
    Ahangari C, Bidabadi MM, Asnaashari M et al (2017) Comparasion of the antimicrobial efficacy of calcium hydroxide and photodynamic therapy against Enterococcus faecalis and Candida albicans in teeth with periapical lesions: an in vivo study. J Lasers Med Sci 8:72–78CrossRefGoogle Scholar
  31. 31.
    Azizi A, Shademan S, Rezai M et al (2016) Effect of photodynamic therapy with two photosensitizers on Streptococcus mutants: in vitro study. Photodiagn Photodyn Ther 16:66–71CrossRefGoogle Scholar
  32. 32.
    Wang Y, Huang Y-Y, Wang Y et al (2017) Photobiomodulation of human adipose-derived stem cells using 810 nm and 980 nm lasers operates via different mechanisms of action. Biochim Biophys Acta 1861:441–449CrossRefGoogle Scholar
  33. 33.
    Costa AF, Assis JCL (2012) In vitro assessment of the bactericidal effect of low-power arsenium-gallium (AsGa) laser treatment. An Bras Dermatol 87:654–656CrossRefGoogle Scholar
  34. 34.
    Pereira PR, Paula JB, Cielinski J et al (2014) Effects of low-intensity laser in in vitro bacterial culture and in vivo infected wounds. Rev Col Bras Cir 41:49–55CrossRefGoogle Scholar
  35. 35.
    Tanaka M, Kinoshita M, Yoshihara Y et al (2012) Optimal photosensitizers for photodynamic therapy of infections should kill bacteria but spare neutrophils. Photochem Photobiol 88:227–232CrossRefGoogle Scholar
  36. 36.
    Mito T, Suzuki T, Kobayasbi T et al (2012) Effect of photodynamic therapy with methylene blue on Acanthamoeba in vitro. Invest Ophthalmol Vis Sci 53:6305–6313CrossRefGoogle Scholar
  37. 37.
    Hamblin MR (2017) Mechanisms and applications of the anti-inflammatory effects of photobiomodulation. AIMS Biophys 4:337–361CrossRefGoogle Scholar

Copyright information

© Springer-Verlag London Ltd., part of Springer Nature 2018

Authors and Affiliations

  • Thais Ferreira Gomes
    • 1
  • Matheus Masalskiene Pedrosa
    • 1
  • Ana Claudia Laforga de Toledo
    • 1
  • Veridiana Wanshi Arnoni
    • 2
  • Mirian dos Santos Monteiro
    • 1
  • Davi Cury Piai
    • 1
  • Silvia Helena Zacarias Sylvestre
    • 1
  • Bruno Ferreira
    • 1
    Email author
  1. 1.Department of Physical therapyUniversity Center UNIFAFIBEBebedouroBrazil
  2. 2.Study Center CollucciCollucci’s ClinicalRibeirão PretoBrazil

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