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Photodynamic inactivation of Paracoccidioides brasiliensis helps the outcome of oral paracoccidiodomycosis

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Abstract

The antifungal drug therapy often employed to treat paracoccidiodomycosis (PCM), an important neglected fungal systemic infection, leads to offensive adverse effects, besides being very long-lasting. In addition, PCM compromises the oral health of patients by leading to oral lesions that are very painful and disabling. In that way, photodynamic therapy (PDT) arises as a new promising adjuvant treatment for inactivating Paracoccidioides brasiliensis (Pb), the responsible fungus for PCM, and also for helping the patients to deal with such debilitating oral lesions. PDT has been linked to an improved microbial killing, also presenting the advantage of not inducing immediate microbial resistance such as drugs. For the present study, we investigated the generation of reactive oxygen species (ROS) by using the fluorescent probes hydroxyphenyl fluorescein (HPF) and aminophenyl fluorescein (APF) after toluidine blue (TBO—37.5 mg/L)-mediated PDT (660 nm, 40 mW, and 0.04 cm2 spot area) and the action of TBO-PDT upon Pb cultures grown for 7 or 15 days in semisolid Fava Netto’s culture medium; we also targeted oral PCM manifestations by reporting the first clinical cases (three patients) to receive topic PDT for such purpose. We were able to show a significant generation of hydroxyl radicals and hypochlorite after TBO-PDT with doses around 90 J/cm2; such ROS generation was particularly useful to attack and inactivate Pb colonies at 7 and 15 days. All three patients reported herein related an immediate relief when it came to pain, mouth opening, and also the ability to chew and swallow. As extracted from our clinical results, which are in fact based on in vitro outcomes, TBO-PDT is a very safe, inexpensive, and promising therapy for the oral manifestations of PCM.

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References

  1. Teixeira Mde M, Theodoro RC, Oliveira FF, Machado GC, Hahn RC, Bagagli E, San-Blas G, Soares Felipe MS (2014) Paracoccidioides lutzii sp. nov.: biological and clinical implications. Med Mycol 52(1):19–28. doi:10.3109/13693786.2013.794311

    PubMed  Google Scholar 

  2. Ramos ESM, Saraiva Ldo E (2008) Paracoccidioidomycosis. Dermatol Clin 26(2):257–269 . doi:10.1016/j.det.2007.11.005 vii

    Article  Google Scholar 

  3. de Oliveira HC, Assato PA, Marcos CM, Scorzoni L, de Paula ESAC, Da Silva JF, Singulani Jde L, Alarcon KM, Fusco-Almeida AM, Mendes-Giannini MJ (2015) Paracoccidioides-host interaction: an overview on recent advances in the paracoccidioidomycosis. Front Microbiol 6:1319. doi:10.3389/fmicb.2015.01319

    PubMed  PubMed Central  Google Scholar 

  4. Mendonca JA, Peron Filho F, Schincariol NB, Vierhout CV, Provenza JR (2016) Musculoskeletal ultrasound findings in paracoccidioidomycosis. Rev Bras Reumatol Engl Ed 56(1):75–78. doi:10.1016/j.rbre.2014.02.022

    Article  PubMed  Google Scholar 

  5. Shikanai-Yasuda MA, Telles Filho Fde Q, Mendes RP, Colombo AL, Moretti ML (2006) Guidelines in paracoccidioidomycosis. Rev Soc Bras Med Trop 39(3):297–310

    Article  PubMed  Google Scholar 

  6. Buitrago MJ, Cuenca-Estrella M (2012) Current epidemiology and laboratory diagnosis of endemic mycoses in Spain. Enferm Infecc Microbiol Clin 30(7):407–413. doi:10.1016/j.eimc.2011.09.014

    Article  PubMed  Google Scholar 

  7. Sperandio FF, Giudice FS, Coelho AP, de Sousa SC, Martins MT (2012) Difficulty in diagnosing oral paracoccidioidomycosis after topical nystatin usage. Gen Dent 60(1):e44–e46

    PubMed  Google Scholar 

  8. de Oliveira Gondak R, Mariano FV, dos Santos Silva AR, Vargas PA, Lopes MA (2012) Single oral paracoccidioidomycosis mimicking other lesions: report of eight cases. Mycopathologia 173(1):47–52. doi:10.1007/s11046-011-9461-3

    Article  PubMed  Google Scholar 

  9. Pedreira Rdo P, Guimaraes EP, de Carli ML, Magalhaes EM, Pereira AA, Hanemann JA (2014) Paracoccidioidomycosis mimicking squamous cell carcinoma on the dorsum of the tongue and review of published literature. Mycopathologia 177(5–6):325–329. doi:10.1007/s11046-014-9739-3

    Article  PubMed  Google Scholar 

  10. de Carli ML, Cardoso BC, Malaquias LC, Nonogaki S, Pereira AA, Sperandio FF, Hanemann JA (2015) Serum antibody levels correlate with oral fungal cell numbers and influence the patients’ response to chronic paracoccidioidomycosis. Mycoses 58(6):356–361. doi:10.1111/myc.12325

    Article  PubMed  Google Scholar 

  11. do Prado Gomes Pedreira R, de Carli ML, Beijo LA, Nonogaki S, Pereira AA, Junior NV, Sperandio FF, Hanemann JA (2016) Oral paracoccidioidomycosis granulomas are predominantly populated by CD163+ multinucleated giant cells. Mycopathologia. doi:10.1007/s11046-016-0016-5

    PubMed  Google Scholar 

  12. Goihman-Yahr M, Pereira J, Isturiz G, Viloria N, Carrasquero M, Saavedra N, de Gomez MH, Roman A, San Martin B, Bastardo de Albornoz MC et al (1992) Relationship between digestive and killing abilities of neutrophils against Paracoccidioides brasiliensis. Mycoses 35(11–12):269–274

    CAS  PubMed  Google Scholar 

  13. Kurita N, Oarada M, Ito E, Miyaji M (1999) Antifungal activity of human polymorphonuclear leucocytes against yeast cells of Paracoccidioides brasiliensis. Med Mycol 37(4):261–267

    Article  CAS  PubMed  Google Scholar 

  14. Meneses-Garcia A, Mosqueda-Taylor A, Morales-de la Luz R, Rivera LM (2002) Paracoccidioidomycosis: report of 2 cases mimicking squamous cell carcinoma. Oral Surg Oral Med Oral Pathol Oral Radiol Endod 94(5):609–613

    Article  PubMed  Google Scholar 

  15. Cock AM, Cano LE, Velez D, Aristizabal BH, Trujillo J, Restrepo A (2000) Fibrotic sequelae in pulmonary paracoccidioidomycosis: histopathological aspects in BALB/c mice infected with viable and non-viable Paracoccidioides brasiliensis propagules. Rev Inst Med Trop Sao Paulo 42(2):59–66

    Article  CAS  PubMed  Google Scholar 

  16. Amaral AC, Fernandes L, Galdino AS, Felipe MS, Soares CM, Pereira M (2005) Therapeutic targets in Paracoccidioides brasiliensis: post-transcriptome perspectives. Genet Mol Res 4(2):430–449

    CAS  PubMed  Google Scholar 

  17. Abadio AK, Kioshima ES, Teixeira MM, Martins NF, Maigret B, Felipe MS (2011) Comparative genomics allowed the identification of drug targets against human fungal pathogens. BMC Genomics 12:75. doi:10.1186/1471-2164-12-75

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Sanglard D, Odds FC (2002) Resistance of Candida species to antifungal agents: molecular mechanisms and clinical consequences. Lancet Infect Dis 2(2):73–85

    Article  CAS  PubMed  Google Scholar 

  19. Baltazar LM, Werneck SM, Soares BM, Ferreira MV, Souza DG, Pinotti M, Santos DA, Cisalpino PS (2015) Melanin protects Paracoccidioides brasiliensis from the effects of antimicrobial photodynamic inhibition and antifungal drugs. Antimicrob Agents Chemother 59(7):4003–4011. doi:10.1128/AAC.04917-14

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Hamblin MR, Hasan T (2004) Photodynamic therapy: a new antimicrobial approach to infectious disease? Photochem Photobiol Sci 3(5):436–450. doi:10.1039/b311900a

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Almeida LM, Zanoelo FF, Castro KP, Borissevitch IE, Soares CM, Goncalves PJ (2012) Cell survival and altered gene expression following photodynamic inactivation of Paracoccidioides brasiliensis. Photochem Photobiol 88(4):992–1000. doi:10.1111/j.1751-1097.2012.01112.x

    Article  CAS  PubMed  Google Scholar 

  22. Sperandio FF, Sharma SK, Wang M, Jeon S, Huang YY, Dai T, Nayka S, de Sousa SC, Chiang LY, Hamblin MR (2013) Photoinduced electron-transfer mechanisms for radical-enhanced photodynamic therapy mediated by water-soluble decacationic C(7)(0) and C(8)(4)O(2) Fullerene Derivatives. Nanomedicine 9(4):570–579. doi:10.1016/j.nano.2012.09.005

    Article  CAS  PubMed  Google Scholar 

  23. Setsukinai K, Urano Y, Kakinuma K, Majima HJ, Nagano T (2003) Development of novel fluorescence probes that can reliably detect reactive oxygen species and distinguish specific species. J Biol Chem 278(5):3170–3175. doi:10.1074/jbc.M209264200

    Article  CAS  PubMed  Google Scholar 

  24. Fava Netto C (1961) Contribuição para o estudo imunológico da blastomicose de Lutz. Revista do Instituto Adolfo Lutz 21:99–194

    Google Scholar 

  25. Singer-Vermes LM, Burger E, Franco MF, Di-Bacchi MM, Mendes-Giannini MJ, Calich VL (1989) Evaluation of the pathogenicity and immunogenicity of seven Paracoccidioides brasiliensis isolates in susceptible inbred mice. J Med Vet Mycol 27(2):71–82

    Article  CAS  PubMed  Google Scholar 

  26. de Castro MS, Ribeiro NV Jr, de Carli ML, Costa Pereira AA, Sperandio FF, Costa Hanemann JA (2016) Photodynamically dealing with bisphosphonate-related osteonecrosis of the jaw: successful case reports. Photodiagn Photodyn Ther. doi:10.1016/j.pdpdt.2016.08.007

    Google Scholar 

  27. Ribeiro CM, Caixeta CA, de Carli ML, Sperandio FF, de Sa Magalhaes EM, Costa Pereira AA, Costa Hanemann JA (2016) Photodynamic inactivation of oral paracoccidioidomycosis affecting woman with systemic lupus erythematosus: an unusual case report. Photodiagn Photodyn Ther. doi:10.1016/j.pdpdt.2016.12.006

    Google Scholar 

  28. Kurokawa CS, Lopes CR, Sugizaki MF, Kuramae EE, Franco MF, Peracoli MT (2005) Virulence profile of ten Paracoccidioides brasiliensis isolates: association with morphologic and genetic patterns. Rev Inst Med Trop Sao Paulo 47(5):257–262. doi:10.1590/S0036-46652005000500004

    Article  PubMed  Google Scholar 

  29. McGowan KL, Buckley HR (1984) Growth characteristics of the yeast phase of Paracoccidioides brasiliensis in a chemically defined medium. J Gen Microbiol 130(11):2797–2801. doi:10.1099/00221287-130-11-2797

    CAS  PubMed  Google Scholar 

  30. Singer-Vermes LM, Ciavaglia MC, Kashino SS, Burger E, Calich VL (1992) The source of the growth-promoting factor(s) affects the plating efficiency of Paracoccidioides brasiliensis. J Med Vet Mycol 30(3):261–264

    Article  CAS  PubMed  Google Scholar 

  31. Restrepo A, Jimenez BE (1980) Growth of Paracoccidioides brasiliensis yeast phase in a chemically defined culture medium. J Clin Microbiol 12(2):279–281

    CAS  PubMed  PubMed Central  Google Scholar 

  32. Berliner MD, Reca ME (1966) Vital staining of Histoplasma capsulatum with Janus Green B. Sabouraudia 5(1):26–29

    Article  CAS  PubMed  Google Scholar 

  33. Castaneda E, Brummer E, Perlman AM, McEwen JG, Stevens DA (1988) A culture medium for Paracoccidioides brasiliensis with high plating efficiency, and the effect of siderophores. J Med Vet Mycol 26(6):351–358

    Article  CAS  PubMed  Google Scholar 

  34. Castano AP, Demidova TN, Hamblin MR (2004) Mechanisms in photodynamic therapy: part one—photosensitizers, photochemistry and cellular localization. Photodiagn Photodyn Ther 1(4):279–293. doi:10.1016/S1572-1000(05)00007-4

    Article  CAS  Google Scholar 

  35. Burger E, Mendes AC, Bani GM, Brigagao MR, Santos GB, Malaquias LC, Chavasco JK, Verinaud LM, de Camargo ZP, Hamblin MR, Sperandio FF (2015) Low-level laser therapy to the mouse femur enhances the fungicidal response of neutrophils against Paracoccidioides brasiliensis. PLoS Negl Trop Dis 9(2):e0003541. doi:10.1371/journal.pntd.0003541

    Article  PubMed  PubMed Central  Google Scholar 

  36. Sperandio FF, Simoes A, Correa L, Aranha AC, Giudice FS, Hamblin MR, Sousa SC (2015) Low-level laser irradiation promotes the proliferation and maturation of keratinocytes during epithelial wound repair. J Biophotonics 8(10):795–803. doi:10.1002/jbio.201400064

    Article  CAS  PubMed  Google Scholar 

  37. Ribeiro CM, de Carli ML, Caixeta CA, Pereira AC, Sperandio FF, Hanemann JA (2017) Photodynamic inactivation of oral paracoccidioidomycosis affecting woman with systemic lupus erythematosus: an unusual case report. Photodiagnosis and Photodynamic Therapy:in press. doi:10.1016/j.pdpdt.2016.12.006

    Google Scholar 

  38. Silva CO, Almeida AS, Pereira AA, Sallum AW, Hanemann JA, Tatakis DN (2007) Gingival involvement in oral paracoccidioidomycosis. J Periodontol 78(7):1229–1234. doi:10.1902/jop.2007.060490

    Article  PubMed  Google Scholar 

  39. Girardi FM, Scroferneker ML, Gava V, Pruinelli R (2012) Head and neck manifestations of paracoccidioidomycosis: an epidemiological study of 36 cases in Brazil. Mycopathologia 173(2–3):139–144. doi:10.1007/s11046-011-9488-5

    Article  CAS  PubMed  Google Scholar 

  40. Hahn RC, Morato Conceicao YT, Santos NL, Ferreira JF, Hamdan JS (2003) Disseminated paracoccidioidomycosis: correlation between clinical and in vitro resistance to ketoconazole and trimethoprim sulphamethoxazole. Mycoses 46(8):342–347

    Article  PubMed  Google Scholar 

  41. Vandeputte P, Ferrari S, Coste AT (2012) Antifungal resistance and new strategies to control fungal infections. Int J Microbiol 2012:713687. doi:10.1155/2012/713687

    Article  PubMed  Google Scholar 

  42. Hu Z, Zhang C, Huang Y, Sun S, Guan W, Yao Y (2012) Photodynamic anticancer activities of water-soluble C(60) derivatives and their biological consequences in a HeLa cell line. Chem Biol Interact 195(1):86–94. doi:10.1016/j.cbi.2011.11.003

    Article  CAS  PubMed  Google Scholar 

  43. Dougherty TJ, Gomer CJ, Henderson BW, Jori G, Kessel D, Korbelik M, Moan J, Peng Q (1998) Photodynamic therapy. J Natl Cancer Inst 90(12):889–905

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  44. Foote CS (1991) Definition of type I and type II photosensitized oxidation. Photochem Photobiol 54(5):659

    Article  CAS  PubMed  Google Scholar 

  45. Takasaki AA, Aoki A, Mizutani K, Schwarz F, Sculean A, Wang CY, Koshy G, Romanos G, Ishikawa I, Izumi Y (2009) Application of antimicrobial photodynamic therapy in periodontal and peri-implant diseases. Periodontol 51:109–140. doi:10.1111/j.1600-0757.2009.00302.x

    Article  Google Scholar 

  46. Sharman WM, Allen CM, van Lier JE (1999) Photodynamic therapeutics: basic principles and clinical applications. Drug Discov Today 4(11):507–517

    Article  CAS  PubMed  Google Scholar 

  47. Marotti J, Sperandio FF, Fregnani ER, Aranha AC, de Freitas PM, Eduardo Cde P (2010) High-intensity laser and photodynamic therapy as a treatment for recurrent herpes labialis. Photomed Laser Surg 28(3):439–444. doi:10.1089/pho.2009.2522

    Article  PubMed  Google Scholar 

  48. Sperandio FF, Marotti J, Aranha AC, Eduardo Cde P (2009) Photodynamic therapy for the treatment of recurrent herpes labialis: preliminary results. Gen Dent 57(4):415–419

    PubMed  Google Scholar 

  49. Sperandio FF, Simoes A, Aranha AC, Correa L, Orsini Machado de Sousa SC (2010) Photodynamic therapy mediated by methylene blue dye in wound healing. Photomed Laser Surg 28(5):581–587. doi:10.1089/pho.2009.2601

    Article  CAS  PubMed  Google Scholar 

  50. Cruz Ede P, Campos L, Pereira Fda S, Magliano GC, Benites BM, Arana-Chavez VE, Ballester RY, Simoes A (2015) Clinical, biochemical and histological study of the effect of antimicrobial photodynamic therapy on oral mucositis induced by 5-fluorouracil in hamsters. Photodiagn Photodyn Ther 12(2):298–309. doi:10.1016/j.pdpdt.2014.12.007

    Article  Google Scholar 

  51. Rocha BA, Melo Filho MR, Simoes A (2016) Antimicrobial Photodynamic Therapy to treat chemotherapy-induced oral lesions: report of three cases. Photodiagn Photodyn Ther 13:350–352. doi:10.1016/j.pdpdt.2015.07.172

    Article  Google Scholar 

  52. Prates RA, Kato IT, Ribeiro MS, Tegos GP, Hamblin MR (2011) Influence of multidrug efflux systems on methylene blue-mediated photodynamic inactivation of Candida albicans. J Antimicrob Chemother 66(7):1525–1532. doi:10.1093/jac/dkr160

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  53. Romanova NA, Brovko LY, Moore L, Pometun E, Savitsky AP, Ugarova NN, Griffiths MW (2003) Assessment of photodynamic destruction of Escherichia coli O157:H7 and Listeria monocytogenes by using ATP bioluminescence. Appl Environ Microbiol 69(11):6393–6398

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  54. Jacobson ES, Tinnell SB (1993) Antioxidant function of fungal melanin. J Bacteriol 175(21):7102–7104

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  55. Kwon-Chung KJ, Rhodes JC (1986) Encapsulation and melanin formation as indicators of virulence in Cryptococcus neoformans. Infect Immun 51(1):218–223

    CAS  PubMed  PubMed Central  Google Scholar 

  56. Dixon DM, Polak A, Szaniszlo PJ (1987) Pathogenicity and virulence of wild-type and melanin-deficient Wangiella dermatitidis. J Med Vet Mycol 25(2):97–106

    Article  CAS  PubMed  Google Scholar 

  57. Wheeler MH, Bell AA (1988) Melanins and their importance in pathogenic fungi. Curr Top Med Mycol 2:338–387

    Article  CAS  PubMed  Google Scholar 

  58. Williamson PR, Wakamatsu K, Ito S (1998) Melanin biosynthesis in Cryptococcus neoformans. J Bacteriol 180(6):1570–1572

    CAS  PubMed  PubMed Central  Google Scholar 

  59. Taborda CP, da Silva MB, Nosanchuk JD, Travassos LR (2008) Melanin as a virulence factor of Paracoccidioides brasiliensis and other dimorphic pathogenic fungi: a minireview. Mycopathologia 165(4–5):331–339

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  60. Nosanchuk JD, Casadevall A (2003) The contribution of melanin to microbial pathogenesis. Cell Microbiol 5(4):203–223

    Article  CAS  PubMed  Google Scholar 

  61. Prates RA, Fuchs BB, Mizuno K, Naqvi Q, Kato IT, Ribeiro MS, Mylonakis E, Tegos GP, Hamblin MR (2013) Effect of virulence factors on the photodynamic inactivation of Cryptococcus neoformans. PLoS One 8(1):e54387. doi:10.1371/journal.pone.0054387

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  62. Silva MB, Thomaz L, Marques AF, Svidzinski AE, Nosanchuk JD, Casadevall A, Travassos LR, Taborda CP (2009) Resistance of melanized yeast cells of Paracoccidioides brasiliensis to antimicrobial oxidants and inhibition of phagocytosis using carbohydrates and monoclonal antibody to CD18. Mem Inst Oswaldo Cruz 104(4):644–648

    Article  PubMed  Google Scholar 

  63. Lewis K (1999) Multidrug resistance: versatile drug sensors of bacterial cells. Curr Biol 9(11):R403–R407

    Article  CAS  PubMed  Google Scholar 

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Acknowledgements

The authors wish to thank the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), grant INCT of Redox Processes in Biomedicine–Redoxoma, and grant #304827/2012-6; Fundação de Amparo à Pesquisa do Estado de Minas Gerais (FAPEMIG), grant CBB-PPM #00119-14, and Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) for the grant AUX PE PNPD 2386/2011 and for the Master’s scholarship provided to Ms. Mendes A.C.

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Correspondence to Felipe F. Sperandio.

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As stated in the “Material and Methods” section, the present study received an approval of the Human Research Ethics Committee of our institution (Protocol # 968.594).

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As stated in the “Material and Methods” section, the clinical treatment of human subjects was only possible after obtaining the Ethics Committee approval and informed consents. In addition, all experiments were performed obeying the relevant laws and institutional guidelines according to the ethical principles of the Declaration of Helsinki.

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dos Santos, L.F.M., Melo, N.B., de Carli, M.L. et al. Photodynamic inactivation of Paracoccidioides brasiliensis helps the outcome of oral paracoccidiodomycosis. Lasers Med Sci 32, 921–930 (2017). https://doi.org/10.1007/s10103-017-2193-y

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