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Targeted photodynamic therapy of established soft-tissue infections in mice

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Abstract

The worldwide rise in antibiotic resistance necessitates the development of novel antimicrobial strategies. Although many workers have used photodynamic therapy (PDT) to kill bacteria in vitro, the use of this approach has seldom been reported in vivo in animal models of infection. We have previously described the first use of PDT to treat excisional wound infections by Gram-(-) bacteria in living mice. However, these infected wound models involved a short timespan between infection (30 min) and treatment by PDT. We now report on the use of PDT to treat an established soft-tissue infection in mice. We used Staphylococcus aureus stably transformed with a Photorhabdus luminescenslux operon (luxABCDE) that was genetically modified to be functional in Gram-(+) bacteria. These engineered bacteria emitted bioluminescence, allowing the progress of the infection to be monitored in both space and time with a low light imaging charge-coupled device (CCD) camera. One million cells were injected into one or both thigh muscles of mice that had previously been rendered neutropenic by cyclophosphamide administration. Twenty-four hours later, the bacteria had multiplied more than one hundredfold; poly-l-lysine chlorin e6 conjugate or free chlorin e6 was injected into one area of infected muscle and imaged with the CCD camera. Thirty minutes later, red light from a diode laser was delivered as a surface spot or by interstitial fiber into the infection. There was a light dose dependent loss of bioluminescence (to <5% of that seen in control infections) not seen in untreated infections or those treated with light alone, but in some cases, the infection recurred. Treatment with conjugate alone led to a lesser reduction in bioluminescence. Infections treated with free chlorin e6 responded less well and the infection subsequently increased over the succeeding days, probably due to PDT-mediated tissue damage. PDT-treated infected legs healed better than legs with untreated infections. This data shows that PDT may have applications in drug-resistant soft-tissue infections.

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References

  1. T. J. Dougherty, C. J. Gomer, B. W. Henderson, G. Jori, D. Kessel, M. Korbelik, J. Moan and Q. Peng, Photodynamic therapy, J. Natl. Cancer Inst., 1998, 90, 889–905.

    Article  CAS  Google Scholar 

  2. H. Von Tappeiner and A. Jodlbauer, Ueber wirkung der photodynamischen (fluorescierenden) Stoffe auf Protozoan und enzyme, Dtsch. Arch. Klin. Med., 1904, 80, 422–487.

    Google Scholar 

  3. M. Wainwright, Photodynamic antimicrobial chemotherapy (PACT), J. Antimicrob. Chemother., 1998, 42, 13–28.

    Article  CAS  Google Scholar 

  4. A. R. Hinman, Global progress in infectious disease control, Vaccine, 1998, 16, 1116–1121.

    Article  CAS  Google Scholar 

  5. M. Michel and L. Gutmann, Methicillin-resistant Staphylococcus aureus and vancomycin-resistant enterococci: therapeutic realities and possibilities, Lancet, 1997, 349, 1901–1906.

    Article  CAS  Google Scholar 

  6. M. Wilson and C. Yianni, Killing of methicillin-resistant Staphylococcus aureus by low-power laser light, J. Med. Microbiol., 1995, 42, 62–66.

    Article  CAS  Google Scholar 

  7. M. Wainwright, D. A. Phoenix, S. L. Laycock, D. R. Wareing and P. A. Wright, Photobactericidal activity of phenothiazinium dyes against methicillin-resistant strains of Staphylococcus aureus, FEMS Microbiol. Lett., 1998, 160, 177–181.

    Article  CAS  Google Scholar 

  8. N. S. Soukos, L. A. Ximenez-Fyvie, M. R. Hamblin, S. S. Socransky and T. Hasan, Targeted antimicrobial photochemotherapy, Antimicrob. Agents Chemother., 1998, 42, 2595–2601.

    Article  CAS  Google Scholar 

  9. M. R. Hamblin, D. A. O’Donnell, N. Murthy, K. Rajagopalan, N. Michaud, M. E. Sherwood and T. Hasan, Polycationic photosensitizer conjugates: effects of chain length and Gram classification on the photodynamic inactivation of bacteria, J. Antimicrob. Chemother., 2002, 49, 941–951.

    Article  CAS  Google Scholar 

  10. M. R. Hamblin, D. A. O’Donnell, N. Murthy, C. H. Contag and T. Hasan, Photodynamic treatment of wound infections in vivo monitored by luminescence imaging, Photochem. Photobiol., 2002, 75, 51–57.

    Article  CAS  Google Scholar 

  11. H. L. Rocchetta, C. J. Boylan, J. W. Foley, P. W. Iversen, D. L. LeTourneau, C. L. McMillian, P. R. Contag, D. E. Jenkins and T. R. Parr, Jr., Validation of a noninvasive, real-time imaging technology using bioluminescent escherichia coli in the neutropenic mouse thigh model of infection, Antimicrob. Agents Chemother., 2001, 45, 129–137.

    Article  CAS  Google Scholar 

  12. C. H. Contag, P. R. Contag, J. I. Mullins, S. D. Spilman, D. K. Stevenson and D. A. Benaron, Photonic detection of bacterial pathogens in living hosts, Mol. Microbiol., 1995, 18, 593–603.

    Article  CAS  Google Scholar 

  13. M. R. Hamblin, T. Zahra, C. H. Contag, A. T. McManus and T. Hasan, Optical monitoring and treatment of potentially lethal wound infections in vivo, J. Infect. Dis., 2003, 187, 1717–1725.

    Article  Google Scholar 

  14. M. R. Hamblin, D. A. O’Donnell, N. Murthy, C. H. Contag and T. Hasan, Rapid control of wound infections by targeted photodynamic therapy monitored by in vivo bioluminescence imaging, Photochem. Photobiol., 2002, 75, 51–57.

    Article  CAS  Google Scholar 

  15. P. Phonimdaeng, M. O’Reilly, P. Nowlan, A. J. Bramley and T. J. Foster, The coagulase of Staphylococcus aureus 8325–4. Sequence analysis and virulence of site-specific coagulase-deficient mutants, Mol. Microbiol., 1990, 4, 393–404.

    Article  CAS  Google Scholar 

  16. K. P. Francis, D. Joh, C. Bellinger-Kawahara, M. J. Hawkinson, T. F. Purchio and P. R. Contag, Monitoring bioluminescent Staphylococcus aureus infections in living mice using a novel luxABCDE construct, Infect. Immun., 2000, 68, 3594–3600.

    Article  CAS  Google Scholar 

  17. K. P. Francis, J. Yu, C. Bellinger-Kawahara, D. Joh, M. J. Hawkinson, G. Xiao, T. F. Purchio, M. G. Caparon, M. Lipsitch and P. R. Contag, Visualizing pneumococcal infections in the lungs of live mice using bioluminescent Streptococcus pneumoniae transformed with a novel gram-positive lux transposon, Infect. Immun., 2001, 69, 3350–3358.

    Article  CAS  Google Scholar 

  18. O. Zak and T. O’Reilly, Animal infection models and ethics: the perfect infection model, J. Antimicrob. Chemother., 1993, 31, 193–205.

    Article  Google Scholar 

  19. J. W. van der Meer, M. Barza, S. M. Wolff and C. A. Dinarello, A low dose of recombinant interleukin 1 protects granulocytopenic mice from lethal gram-negative infection, Proc. Natl. Acad. Sci. U. S. A., 1988, 85, 1620–1623.

    Article  Google Scholar 

  20. N. Komerik, M. Wilson and S. Poole, The effect of photodynamic action on two virulence factors of gram-negative bacteria, Photochem. Photobiol., 2000, 72, 676–680.

    Article  CAS  Google Scholar 

  21. F. Berthiaume, S. Reiken, M. Toner, R. Tompkins and M. Yarmush, Antibody-targeted photolysis of bacteria in vivo, Biotechnology, 1994, 12, 703–706.

    CAS  PubMed  Google Scholar 

  22. G. F. Lombard, S. Tealdi and M. M. Lanotte, The treatment of neurosurgical infections by lasers and porphyrins, in Photodynamic Therapy of Tumors and other Diseases, ed. G. Jori and C. A. Perria, Edizione Libreria Progetto, Padova, 1985, pp. 363–366.

    Google Scholar 

  23. C. Perry, Methicillin-resistant Staphylococcus aureus, J. Wound Care, 1996, 5, 31–34.

    Article  CAS  Google Scholar 

  24. T. L. Smith, M. L. Pearson, K. R. Wilcox, C. Cruz, M. V. Lancaster, B. Robinson-Dunn, F. C. Tenover, M. J. Zervos, J. D. Band, E. White and W. R. Jarvis, Emergence of vancomycin resistance in Staphylococcus aureus. Glycopeptide-Intermediate Staphylococcus aureus Working Group, N. Engl. J. Med., 1999, 340, 493–501.

    Article  CAS  Google Scholar 

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Correspondence to Michael R. Hamblin.

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Gad, F., Zahra, T., Francis, K.P. et al. Targeted photodynamic therapy of established soft-tissue infections in mice. Photochem Photobiol Sci 3, 451–458 (2004). https://doi.org/10.1039/b311901g

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