Methylene blue-mediated photodynamic therapy in the treatment of oral microbiota. A Systematic Review




Photochemotherapy; Photosensitizer; Mouth; Biofilms.


Objective: The objective of the present review was to relate the scientific production presented in the last 5 years on the effect of photodynamic therapy using the methylene blue photosensitizer on the microorganisms present in the oral biofilm. Methods: systematic review looking for research in English of primary studies such as randomized clinical trials and without randomization, individual case-control, and qualitative descriptive studies, using the PubMed/MEDLINE and Scopus databases as references, from 2017 to 2021. Results: 15 articles were analyzed, being 67% national publications and 33% international studies. Considering the samples described in the studies, 73% were in vitro, 7% were in vivo and 20% were clinical samples. As for the light source, 66% used laser, 20% used LED and 14% the association of laser and LED devices. The power density used ranged from 0.0026 W/cm2 to 1415 W/cm2. The energy density ranged from 3 J/cm2 to 640 J/cm2. The power ranged from 10 mW to 110 mW. The irradiation time ranged from 24 to 294 seconds. In most publications, 87% used the wavelength of 660 nm. As for the photosensitizer, the most used concentration, in 33% of the studies, was 0.01%, with the pre-irradiation time most used being 300 seconds in 40% of the studies. Conclusion: Photodynamic Methylene Blue Therapy is being accepted as effective in therapies of various oral curative conditions and has been shown to be a useful and effective therapy, as well as a complementary approach to control the development/growth of oral biofilm in clinical situations.


Azizi, A., Amirzadeh, Z., Rezai, M., Lawaf, S., & Rahimi, A. (2016). Effect of photodynamic therapy with two photosensitizers on Candida albicans. Journal of Photochemistry & Photobiology, B: Biology, 158, 267–273. DOI: 10.1016/j.jphotobiol.2016.02.027.

Azizi, A., Shohratia, P., Goudarzi, M., Lawaf, S., & Rahimi, A. (2019). Comparison of the effect of photodynamic therapy with curcumin and methylene Blue on streptococcus mutans bacterial colonies. Photodiagnosis and Photodynamic Therapy, 27, 203–209. DOI: 10.1016/j.pdpdt.2019.06.002.

Bandara, H. M., Matsubara, V. H., & Samaranayake, L. P. (2017). Future therapies targeted towards eliminating Candida biofilms and associated infections. Expert Review of Anti-infective Therapy, 15(3), 299–318.

Bevilacqua, I. M., Nicolau, R. A., Khouri, S., Brugnera Jr., A., Teodoro, G. R., Zangaro, R. A., & Pacheco, M.T. (2007). The impact of photodynamic therapy on the viability of Streptococcus mutans in a planktonic culture. Photomed. Laser Surg.,513–518.

Borsatto, M. C., Correa-Afonso, A. M., Lucisano, M. P., Bezerra da Silva, R. A., Paula-Silva, F. W., Nelson-Filho, P., & Bezerra da Silva, L. A. (2016). One-session root canal treatment with antimicrobial photodynamic therapy (aPDT): an in vivo study, Int Endod J, 49, 511–518. DOI: 10.1111/iej.12486.

Burns, P. B., Rohrich, R. J., & Chung, K. C. (2011). The levels of evidence and their role in evidence-based medicine. Plastic and Reconstructive Surgery, 128(1), 305.

Colina, G. A., Freire, F., Santos, T. P. C., Sobrinho, N. G., Aquino, S., Prates, R. A., Silva, D. F. T., Horliana, A. C. R. T., & Pavani, C. (2018). Controlling Methylene Blue aggregation: a more eficiente alternative to treat Candida albicans infections using Photodynamic Therapy. Photochemical & Photobiological Sciences, 17(10), 1355-1364. DOI: 10.1039/C8PP00238J

Daliri, F., Azizi, A., Goudarzi, M., Lawaf, S., & Rahimi, A. (2019). In vitro comparison of the effect of photodynamic therapy with curcumin and methylene blue on Candida albicans colonies. Photodiagnosis and Photodynamic Therapy, 26, 193-198. DOI: 10.1016/j.pdpdt.2019.03.017.

De Oliveira, A. B., Ferrisse, T. M., Marques, R. S., De Annunzio, S. R., Brighenti, F. L., & Fontana, C.R. (2019). Effect of Photodynamic Therapy on Microorganisms Responsible for Dental Caries: A Systematic Review and Meta-Analysis. Int J Mol Sci, 20(14), 3585. DOI: 10.3390/ijms20143585.

Diniz, I.M., Horta, I.D., Azevedo, C.S., Elmadjian, T.R., Matos, A.B., Simionato, M.R., & Marques, M.M. (2015). Antimicrobial photodynamic therapy: A promise candidate for caries lesions treatment. Photodiagnosis Photodyn Ther, 12, 511–518.

Di Poto, A., Sbarra, M. S., Provenza, G., Visai, L., & Speziale, P. (2009). The effect of photodynamic treatment combined with antibiotic action or host defence mechanisms on Staphylococcus aureus biofilms. Biomaterials, 30(18), 3158–3166.

Eduardo, F. P., Bezinelli, L. M., Gobbi, M. F., Santos, V.M., Maluf, F.C., & Correa, L. (2019). Severe oral infection caused by Pseudomonas aeruginosa effectively treated with methylene blue-mediated photodynamic inactivation. Photodiagnosis and Photodynamic Therapy, 26, 284–286. DOI: 10.1016/j.pdpdt.2019.04.013.

Filipini, S. M. R., Campagnolo, C. B., Dutra, D. A. M., Maciel, R. M., Danesi, C. C., & Kantorski, K. Z. (2019). Adjunctive antimicrobial photodynamic therapy using methylene blue/ethanol formulation in experimental periodontitis in diabetic rats: short-term results. Lasers in Medical Science, 34(6), 1253-1260. DOI: 10.1007/s10103-019-02733-4.

Fonseca, M. B., Júnior, P. O., Pallota, R. C., Filho, H. F., Denardin, O. V., Rapoport, A., Dedivitis, R. A., Veronezi, J. F., Genovese, W. J., & Ricardo, A. L. (2008). Photodynamic therapy for root canals infected with Enterococcus faecalis. Photomedicine and Laser Surgery, 26(3), 209-213.

Fontana, C. R., Abernethy, A. D., Som, S., Ruggiero, K., Doucette, S., Marcantonio, R. C., Boussios, C. I., Kent, R., Goodson, J. M., Tanner, A. C. R., & Soukos, N. S. (2009). The antibacterial effect of photodynamic therapy in dental plaque-derived biofilms. Journal of Periodontal Research, 44(6), 751-759.

Freitas, L. M., Lorenzon, E. N., Cillid, E. M., Oliveira, K. T., Fontana, C. R., & Mang, T. S. (2019). Photodynamic and peptide-based strategy to inhibit Gram-positive bacterial biofilm formation. Biofouling, 35(7), 742-757. DOI:10.1080/08927014.2019.1655548.

Freitas, L. S. F., Rossoni, R. D., Jorge, A. O. C., & Junqueira, J. C. (2017). Repeated applications of photodynamic therapy on Candida glabrata biofilms formed in acrylic resin polymerized. Lasers in Medical Science, 32(3), 549-555. DOI 10.1007/s10103-017-2147-4.

Fumes, A. C., Romualdo, P. C., Monteiro, R. M., Watanabe, E., Corona, S. A. M., & Borsatto, M. C. (2018). Influence of pre-irradiation time employed in antimicrobial photodynamic therapy with diode laser. Lasers in Medical Science, 33, 67–73. DOI 10.1007/s10103-017-2336-1.

Fumes, A. C., Telles, P. D. S., Corona, S. A. M., & Borsatto, M. C. (2018). Effect of aPDT on Streptococcus mutans and Candida albicans present in the dental biofilm: Systematic review. Photodiagnosis Photodyn Ther, 21, 363-366. DOI: 10.1016/j.pdpdt.2018.01.013.

Furtado, G. S., Paschoal, M. A. B., Grenho, L. C. S., & Lago, A. D. N. (2020). Does pre-irradiation time influence the efficacy of antimicrobial photodynamic therapy? Photodiagnosis and Photodynamic Therapy, 31, 101884. DOI:

Gad, F., Zahra, T., Hasan, T., & Hamblin, M. R. (2004). Effects of growth phase and extracellular slime on photodynamic inactivation of Gram-positive pathogenic bacteria. Antimicrobial Agents and Chemotherapy, 48(6), 2173-2178.

Garcez, A. S., Nunez, S. C., Azambuja, Jr N., Fregnani, E. R., Rodriguez, H. M. H., Hamblin, M. R., Suzuki, H., & Ribeiro, M. S. (2013). Effects of Photodynamic Therapy on Gram-Positive and Gram-Negative Bacterial Biofilms by Bioluminescence Imaging and Scanning Electron Microscopic Analysis. Photomedicine and Laser Surgery, 31(11), 519-525. DOI: 10.1089/pho.2012.3341

George S, Kishen A. (2007). Photophysical, photochemical, and photobiological characterization of methylene blue formulations for light-activated root canal disinfection. J Biomed Opt, 12:29–34.

Gleiznys, A., Zdanaviciene, E., & Zilinskas, J. (2015). Candida albicans importance to denture wearers. A literature review. Stomatologija, 17, 54–66.

Ghorbani, J., Rahban, D., Aghamiri, S., Teymouri, A. & Bahador, A. (2018). Photosensitizers in antibacterial photodynamic therapy: an overview. Laser Ther, 27(4), 293-302. doi: 10.5978/islsm.27_18-RA-01.

Guglielmi C. de A., Simionato, M. R., Ramalho, K. M., Imparato, J. C., Pinheiro, S. L., & Luz, M. A. (2011). Clinical use of photodynamic antimicrobial chemotherapy for the treatment of deep carious lesions. J Biomed Opt, 16, 088003. doi: 10.1117/1.3611009.

Hamblin, M. R. (2016). Antimicrobial photodynamic inactivation: a bright new technique to kill resistant microbes. Current Opinion in Microbiology, 33, 67–73.

Hu, X., Huang, Y. Y., Wang, Y., Wang, X., & Hamblin, M. R. (2018). Antimicrobial Photodynamic Therapy to Control Clinically Relevant Biofilm Infections. Frontiers in Microbiology, 27(9), 1299.

Huang, T. C., Chen, C. J., Ding, S. J., & Chen, C. C. (2019). Antimicrobial efficacy of methylene blue-mediated photodynamic therapy on titanium alloy surfaces in vitro. Photodiagnosis and Photodynamic Therapy, 25, 7–16. DOI: 10.1016/j.pdpdt.2018.11.008.

Javali, M. A., AlQahtani, N. A., Ahmad, I., & Ahmad, I. (2019). Antimicrobial photodynamic therapy (light source; methylene blue; titanium dioxide): Bactericidal effects analysis on oral plaque bacteria: An in vitro study. Nigerian Journal of Clinical Practice, 22, 1654-1661. DOI: 10.4103/njcp.njcp_189_19.

Jori, G., Fabris, C., Soncin, M., Ferro, S., Coppellotti, O., Dei, D., Fanetti, L., Chiti, G., & Roncucci, G. (2006). Photodynamic therapy in the treatment of microbial infections: basic principles and perspective applications. Lasers in Surgery and Medicine, 38(5), 468–481. DOI: 10.1002/lsm.20361.

Junqueira, H. C. C., Severino, D., Dias, L. G. G., Gugliotti, M. S. S., & Baptista, M. S. S. (2004). Modulation of methylene blue photochemical properties based on adsorption at aqueous micelle interfaces, Phys. Chem. Chem. Phys., 2320–2328.

Kwiatkowski, S., Knap, B., Przystupski, D., Saczko, J., Kedzierska, E., Knap-Czop, K., Kotlinska, J., Michel, O., Kotowski, K., & Kulbacka, J. (2018). Photodynamic therapy-mechanisms, photosensitizers and combinations. Biomed. Pharm.,106, 1098 - 1107.

Lacerda, M. F. L. S., Alfenas, C. F., & Campos, C. N. (2014). Terapia fotodinâmica associada ao tratamento endodôntico -revisão de literatura. Revista da Faculdade de Odontologia, 19(1), 115-120.

Li, R., Yuan, L., Jia, W., Qin, M., & Wang, Y. (2021) Effects of Rose Bengal and Methylene Blue Mediated Potassium Iodide Potentiated Photodynamic Therapy on Enterococcus faecalis: A Comparative Study. Lasers in Surgery and Medicine, 53(3), 400-410. DOI: 10.1002/lsm.23299.

Li, Y., Wang, B., Zheng, S. & He, Y. (2018). Photodynamic therapy in the treatment of oral leukoplakia: a systematic review. Photodiagnosis and Photodynamic Therapy, 25,17-22.

Lobão, F. R., Viçoso, M. D., Guerreiro, L., Palazzo, M., Almeida, P. & Vargas, G. (2016). O papel da Odontologia Intensiva. Academus Revista Científica da Saúde, 1(3), 1-6.

Maciel, C. M., Piva, M. R., Ribeiro, M. A. G., Santana Santos, T. de, Ribeiro, C. F., & Martins-Filho, P. R. S. (2016). Methylene Blue-Mediated Photodynamic Inactivation Followed by Low-Laser Therapy versus Miconazole Gel in the Treatment of Denture Stomatitis. Journal of Prosthodontics, 25(1), 28–32. DOI: 10.1111/jopr.12284.

Malik, N. K. A., & Alkadhi, O. H. (2020). Effectiveness of mechanical debridement with and without antimicrobial photodynamic therapy against oral yeasts in children with gingivitis undergoing fixed orthodontic therapy. Photodiagnosis and Photodynamic Therapy, 31, 101768. DOI: 10.1016/j.pdpdt.2020.101768.

Meisel, P., Kocher, T. (2005). Photodynamic therapy for periodontal diseases: state of the art. J Photochem Photobiol B, 79, 159-170.

Mendes, K. D. S., Silveira, R. C. D. C. P., & Galvão, C. M. (2008). Revisão integrativa: método de pesquisa para a incorporação de evidências na saúde e na enfermagem. Texto & Contexto Enfermagem, 17(4), 758-764.

Monteiro, C. C. T., Franco, C. B., Feitosa, E. A., De Araújo, M. A. & Brígido, J.A. (2021). Terapia fotodinâmica antimicrobiana: uma modalidade promissora no tratamento da estomatite protética. Brazilian Journal of Health Review, 4(6), 25886-25893. DOI:10.34119/bjhrv4n6-184.

Nagata, J. Y., Hioka, N., Kimura, E., Batistela, V. R., Terada, R. S., Graciano, A. X., Baesso, M. L., & Hayacibara, M. F. (2012). Antibacterial photodynamic therapy for dental caries: evaluation of the photosensitizers used and light source properties. Photodiagnosis and Photodynamic Therapy, 9(2), 122–131.

Nemezio, M. A., Farias, S. S. S., Borsatto, M. C., Aires, C. P., & Corona, S. A. M. (2017). Effect of methylene blue-induced photodynamic therapy on a Streptococcus mutans biofilm model. Photodiagnosis and Photodynamic Therapy, 20, 234–237. DOI: 10.1016/j.pdpdt.2017.10.025.

Ochsner, M. (1997). Photophysical and photobiological processes in the photodynamic therapy of tumours. J Photochem Photobiol B, 39, 1-1825.

Pereira, C. A., Romeiro, R. L., Costa, A. C., Machado, A. K., Junqueira, J. C., & Jorge, A. O. (2011). Susceptibility of Candida albicans, Staphylococcus aureus, and Streptococcus mutans biofilms to photodynamic inactivation: an in vitro study. Lasers Med. Sci., 26, 341–348. DOI: 10.1007/s10103-010-0852-3.

Perkowski, K., Baltaza, W., Conn, D. B., Marczynska-Stolarek, M., Chomicz, L. (2019). Examination of oral biofilm microbiota in patients using fixed orthodontic appliancesin order to prevent risk factors for health complications. Annals of Agricultural and Environmental Medicine, 26(2), 231–235. DOI: 10.26444/aaem/105797.

Pimentel, J. K. P., & Dias, K. S. P. A. Conceitos e aplicabilidade da terapia fotodinâmica na periodontia: revisão de Literatura. (2021). Research, Society and Development, 10(15), e277101522931. DOI:

Pourhajibagher, M., Salehi Vaziri, A., Takzaree, N., & Ghorbanzadeh, R. (2019). Physico-mechanical and antimicrobial properties of an orthodontic adhesive containing cationic curcumin doped zinc oxide nanoparticles subjected to photodynamic therapy. Photodiagnosis Photodyn Ther, 25, 239–246. DOI: 10.1016/j.pdpdt.2019.01.002.

Prochnow, E. P., Martins, M. R., Campagnolo, C. B., Santos, R. C., Villetti, M. A., & Kantorski, K. Z. (2016). Antimicrobial

photodynamic effect of phenothiazinic photosensitizers in formulations with ethanol on Pseudomonas aeruginosa biofilms, Photodiagnosis Photodyn. Ther,13, 291–296. DOI: 10.1016/j.pdpdt.2015.08.008.

Queiroz, A. C., Suaid, F. A., De Andrade, P. F., Oliveira, F. S., Novaes Jr., A. B., Taba Jr., M., Palioto, D. B., Grisi, M. F., & Souza, S. L. (2015). Adjunctive effect of antimicrobial photodynamic therapy to nonsurgical periodontal treatment in smokers: a randomized clinical trial. Lasers Med. Sci., 30, 617–625. DOI: 10.1007/s10103-013-1379-1.

Ramalho, K. M., Cunha, S. R., Mayer-Santos, E., Eduardo, C. P., Freitas, P. M., Aranha, A., & Moura-Netto, C. (2017). In vitro evaluation of methylene blue removal from root canal after Photodynamic Therapy. Photodiagnosis and Photodynamic Therapy, 20, 248-252.

Santezi, C., Reina, B. D., & Dovigo, L. N. (2018). Curcumin-mediated Photodynamic Therapy for the treatment of oral infections- a review. Photodiagnosis and Photodynamic Therapy, 21, 409–415.

Santos, C. M. D. C., Pimenta, C. A. D. M., & Nobre, M. R. C. (2007). The PICO strategy for the research question construction and evidence search. Revista Latino-Americana de Enfermagem, 15(3), 508-511.

Shen, J.J., Jemec, G. B. E., Arendrup, M. C. & Saunte, D. M. L. (2020). Photodynamic therapy treatment of superficial fungal infections: A systematic review. Photodiagnosis Photodyn Ther, 31, 101774. DOI: 10.1016/j.pdpdt.2020.101774.

Shrestha, A., Shi, Z., Neoh, K. G., & Kishen, A. (2010). Nanoparticulates for antibiofilm treatment and effect of aging on its antibacterial activity. Journal of Endodontics, 36(6), 1030–1035.

Simmons, B. J., Griffith, R. D., Falto-Aizpurua, L. A., & Nouri, K. (2015). An Update on photodynamic therapies in the treatment of onychomycosis. Journal of the European Academy Dermatology and Venereology, 29(7), 1275-1279. DOI: 10.1111/jdv.12950.

Siqueira, J. F., & Sen, B. H. (2004). Fungi in endodontic infections. Oral Surgery, Oral Medicine, Oral Pathology, Oral Radiology, and Endodontology, 97(5), 632–641.

Soares, L. G. P., Crugeira, P. J. L., Nunes, I. P. F., Santos, A. S., Cangussú, M. C. T., Almeida, P. F., Pinheiro, A. L. B., & Habib, F. A. L. (2019). Oral microbiological control by photodynamic action in orthodontic patients. Photodiagnosis and Photodynamic Therapy, 28, 221–225.

Stuber, M., Fonseca Filho, P. F. O. da, Alberton, C. S. ., Santos, V. C. dos ., Silva, B. M. da ., Kaizer, M. da R., Wambier, L. M., & Tomazinho, F. S. F. (2021). Does the use of photodynamic therapy compared to calcium hydroxide reduce intracanal microorganisms in teeth with pulp necrosis? a systematic review and meta-analysis. Research, Society and Development, 10(12), e136101220058. DOI: 10.33448/rsd-v10i12.20058.

Takasaki, A. A., Aoki, A., Mizutani, K., Schwarz, F., Sculean, A., Wang, C., & Izumi, Y. (2009). Application of antimicrobial photodynamic therapy in periodontal and peri-implant diseases. Periodontology 2000, 51(1), 109-140.

Teichert, M. C., Jones, J. W., Usacheva, M. N., & Biel, M. A. (2002). Treatment of oral candidiasis with methylene blue-mediated photodynamic therapy in an immunodeficient murine model. Oral Surg Oral Med Oral Pathol Oral Radiol Endod, 93(2), 155–160.

Wadhwa, A., Mallapragada, S., & Sharma, P. (2021). Novel indocyanine green mediated antimicrobial photodynamic therapy in the management of chronic periodontitis - A randomized controlled clinico-microbiological pilot study. Journal of oral biology and craniofacial research, 11(1), 57–62.

Wainwright, M. Methylene blue derivatives - suitable photoantimicrobials for blood product disinfection? (2000). Int J Antimicrob Agents,16(4),381-94.

Wainwright, M. (2005). The development of phenothiazinium photosensitizers. Photodiagnosis and Photodynamic Therapy, 2, 263–272.

Warrier, A., Mazumder, N., Prabhu, S., Satyamoorthy, K., & Murali, T. S. (2021). Photodynamic therapy to control microbial biofilms. Photodiagnosis and Photodynamic Therapy, 33, 102090.

Zago, L. H. P., Annunzio, S. R., Oliveira, K. T., Barbugli, P. A., Barreto, I. M. Q., Feres, M., & Fontana, C. R. (2020). Antimicrobial photodynamic therapy against metronidazol e resistant dental plaque bactéria. Journal of Photochemistry & Photobiology, B: Biology, 209, 111903.




How to Cite

MORETI, L. C. T. .; GARCIA, L. A. .; FERNANDES, K. G. C. .; KOZUSNY-ANDREANI, D. I. .; SOUZA, J. A. S. .; TIM , C. R. . Methylene blue-mediated photodynamic therapy in the treatment of oral microbiota. A Systematic Review. Research, Society and Development, [S. l.], v. 11, n. 6, p. e53411629001, 2022. DOI: 10.33448/rsd-v11i6.29001. Disponível em: Acesso em: 29 may. 2022.



Health Sciences