title: Papain gel containing methylene blue for simultaneous caries removal and antimicrobial photoinactivation against Streptococcus mutans biofilms.
abstract: This study intended to evaluate the effects of a papain-gel with a red-light absorbing pigment (methylene blue - MB) to mediate photodynamic therapy (PDT) against Streptococcus mutans biofilms. The PapaMBlue was compared with free MB to generate reactive oxygen species using fluorescence probes (SOSG and HPF). PDT (660-nm light) was carried out against S. mutans biofilms grown on either plastic dishes or on collagen membrane and assayed by CFU, live-dead staining using confocal microscopy, transmission electron microscopy and H&E staining for collagen films. Cytotoxicity and subcellular localization was studied in human fibroblasts. Sponges of bioabsorbable type I collagen membrane were exposed to papain based gel, irradiated with laser and analyzed about their integrity by ATR-FTIR. The PapaMBlue produced higher amounts of singlet oxygen and hydroxyl radicals than free MB, possibly due to better disaggregation of the dye in solution. The PapaMBlue antimicrobial effects on biofilms proved to be capable of reducing the S. mutans. Both MTT and PrestoBlue assays showed higher cell viability and metabolism scores in fibroblasts treated with PapaMBlue and MB, possibly due to stimulation of mitochondrial activity and that collagen triple helix is unaffected. The PapaMBlue is equally effective as MB in destroying S. mutans biofilms growing on plastic or collagen without affecting fibroblasts.
authors.affiliation: Massachusetts General Hospital, Wellman Center for Photomedicine, Boston, MA 02114, USA.|Harvard Medical School, Department of Dermatology, Boston, MA 02114, USA.|Postgraduate Program in Biophotonics Applied to Health Sciences, Nove de Julho University (UNINOVE), São Paulo, SP, Brazil.
authors.forename: Zenildo Santos
authors.initials: ZS
authors.lastname: Silva
authors.affiliation: Massachusetts General Hospital, Wellman Center for Photomedicine, Boston, MA 02114, USA.|Harvard Medical School, Department of Dermatology, Boston, MA 02114, USA.
authors.forename: Ying-Ying
authors.initials: YY
authors.lastname: Huang
authors.affiliation: Massachusetts General Hospital, Wellman Center for Photomedicine, Boston, MA 02114, USA.|Harvard Medical School, Department of Dermatology, Boston, MA 02114, USA.|Postgraduate Program in Bioengineering of São Paulo University, São Carlos, SP, Brazil.
authors.forename: Lucas Freitas
authors.initials: LF
authors.lastname: de Freitas
authors.affiliation: Postgraduate Program in Biophotonics Applied to Health Sciences, Nove de Julho University (UNINOVE), São Paulo, SP, Brazil.
authors.forename: Cristiane Miranda
authors.initials: CM
authors.lastname: França
authors.affiliation: Postgraduate Program in Biophotonics Applied to Health Sciences, Nove de Julho University (UNINOVE), São Paulo, SP, Brazil.
authors.forename: Sergio Brossi
authors.initials: SB
authors.lastname: Botta
authors.affiliation: Center of Engineering, Modeling and Applied Social Sciences, Federal University of ABC, Sao Bernardo do Campo, SP, Brazil.
authors.forename: Patrícia Aparecida
authors.initials: PA
authors.lastname: Ana
authors.affiliation: Postgraduate Program in Biophotonics Applied to Health Sciences, Nove de Julho University (UNINOVE), São Paulo, SP, Brazil.
authors.forename: Raquel Agnelli
authors.initials: RA
authors.lastname: Mesquita-Ferrari
authors.affiliation: Postgraduate Program in Biophotonics Applied to Health Sciences, Nove de Julho University (UNINOVE), São Paulo, SP, Brazil.
authors.forename: Kristianne Porta
authors.initials: KP
authors.lastname: Santos Fernandes
authors.affiliation: Postgraduate Program in Biophotonics Applied to Health Sciences, Nove de Julho University (UNINOVE), São Paulo, SP, Brazil.
authors.forename: Alessandro
authors.initials: A
authors.lastname: Deana
authors.affiliation: Postgraduate Program in Biophotonics Applied to Health Sciences, Nove de Julho University (UNINOVE), São Paulo, SP, Brazil.
authors.forename: Cintia Raquel
authors.initials: CR
authors.lastname: Lima Leal
authors.affiliation: Postgraduate Program in Biophotonics Applied to Health Sciences, Nove de Julho University (UNINOVE), São Paulo, SP, Brazil.
authors.forename: Renato Araujo
authors.initials: RA
authors.lastname: Prates
authors.affiliation: Massachusetts General Hospital, Wellman Center for Photomedicine, Boston, MA 02114, USA.|Harvard Medical School, Department of Dermatology, Boston, MA 02114, USA.|Harvard-MIT Division of Health Sciences and Technology, Cambridge, MA 02139, USA.
authors.forename: Michael R
authors.initials: MR
authors.lastname: Hamblin
authors.affiliation: Postgraduate Program in Biophotonics Applied to Health Sciences, Nove de Julho University (UNINOVE), São Paulo, SP, Brazil.
authors.forename: Sandra Kalil
authors.initials: SK
authors.lastname: Bussadori
mesh_terms: D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D018441:Biofilms / Q000187:drug effects*; D003731:Dental Caries / Q000188:drug therapy* / Q000382:microbiology; D005782:Gels; D006801:Humans; D017665:Hydroxyl Radical / Q000378:metabolism / Q000494:pharmacology; D008751:Methylene Blue / Q000378:metabolism / Q000494:pharmacology*; D018613:Microscopy, Confocal; D046529:Microscopy, Electron, Transmission; D010206:Papain / Q000378:metabolism / Q000494:pharmacology*; D010778:Photochemotherapy / Q000379:methods; D017319:Photosensitizing Agents / Q000494:pharmacology; D026082:Singlet Oxygen / Q000378:metabolism / Q000494:pharmacology; D013295:Streptococcus mutans / Q000187:drug effects* / Q000502:physiology / Q000648:ultrastructure
chemical_list: D000900:Anti-Bacterial Agents; D005782:Gels; D017319:Photosensitizing Agents; D026082:Singlet Oxygen; D017665:Hydroxyl Radical; D010206:Papain; D008751:Methylene Blue