The effects of photodynamic therapy with blue light and papain-based gel associated with Urucum, on collagen and fibroblasts: a spectroscopic and cytotoxicity analysis

  • Zenildo Santos SilvaJrEmail author
  • Cristiane Miranda França
  • Renato Araújo Prates
  • Sergio Brossi Botta
  • Raquel Agnelli Mesquita Ferrari
  • Patricia Aparecida Ana
  • Christiane Pavani
  • Kristianne Porta Santos Fernandes
  • Daniela de Fátima Teixeira da Silva
  • Michael R. Hamblin
  • Sandra Kalil BussadoriEmail author
Letter to the Editor


Papacarie Duo™ is clinically used and has proven effectiveness; however, it is necessary to improve its antimicrobial action. The combined treatment of Papacarie Duo™ with Urucum (Bixa Orellana) could create a potential tool for dental caries treatment; its extract obtained from the seeds’ pericarp contains a water-soluble primary pigment (cis-bixin) with smaller amounts of other carotenoids. The dicarboxylic acid salts of cis-norbixin and trans-norbixin occur in heated alkaline solutions. To analyze the absorption spectra and cytotoxicity (with human dermal fibroblasts) in different concentrations of Urucum, associated or not with Papacarie Duo™, we performed this in vitro study. The effects of pure Urucum, Papacarie Duo™, and PapaUrucum™ on the microstructure of collagen were also analyzed. The application of papain-based gel with Urucum did not present cytotoxicity, its exhibit UV absorption spectrum peak around 460 ± 20 nm. Also, it showed that the compound used did not alter the chemical structure of collagen. Consequently, this product could be used as a chemomechanical method to remove dentin caries as well as being a potential product for antimicrobial photodynamic therapy (aPDT) application.


Cytotoxicity Papain-based gel Fourier transform infrared spectroscopy Dentin 



The authors gratefully to Nove de Julho University to the  technological support.

Author contributions

ZSSJ, CMF, SBB, and PAA participated in the conception and design of the study, data collection, and drafting of the present manuscript. RAMF, LCS, MRH and MLLG helped draft the manuscript. DFTS performed statistical analyses, contributed to the design of the study. CMF, RAMF, CP, KPSF, and SKB critically reviewed the manuscript for intellectual content. SKB conceived the study and helped draft the manuscript. The final paper was read and approved by all the authors.

Funding information

Z. Santos S. Jr. was finanacially supported by CAPES-Brazil grant 99999.002158/2014-00. M.R. Hamblin was financially supported by US NIH grant R01AI050875.

Compliance with ethical standards

Conflict of interest

The authors declare non-financial interests concerning the work described. M.R. Hamblin is also a present member of the Transdermal Cap, Inc. scientific advisory board.


  1. 1.
    Bussadori SK, Castro LC, Galvão AC (2006) Papain gel: a new chemo-mechanical caries removal agent. J Clin Pediatr Dent 30(2):115–119CrossRefGoogle Scholar
  2. 2.
    Bussadori S, Guedes C, Hermida Bruno M, Ram D (2008) Chemo-mechanical removal of caries in an adolescent patient using a papain gel: case report. J Clin Pediatr Dent 32(3):177–180CrossRefGoogle Scholar
  3. 3.
    Motta LJ, Bussadori SK, Campanelli AP, Silva AL, Alfaya TA, Godoy CHL et al (2014) Randomized controlled clinical trial of long-term chemo-mechanical caries removal using PapacarieTM gel. J Appl Oral Sci 22(4):307–313CrossRefGoogle Scholar
  4. 4.
    Motta LJ, Bussadori SK, Campanelli AP, Silva AL, Alfaya TA, Godoy CHL et al (2014) Efficacy of Papacarie¯ in reduction of residual bacteria in deciduous teeth: a randomized, controlled clinical trial. Clinics. 69(5):319–322CrossRefGoogle Scholar
  5. 5.
    Bohari MR, Chunawalla YK, Ahmed BMN (2012) Clinical evaluation of caries removal in primary teethusing conventional, chemomechanical and lasertechnique: an in vivo study. J Contemp Dent Pract 13(1):40–47CrossRefGoogle Scholar
  6. 6.
    Kochhar GK, Srivastava N, Pandit I, Gugnani N, Gupta M (2011) An evaluation of different caries removal techniques in primary teeth: a comparitive clinical study. J Clin Pediatr Dent 36(1):5–10CrossRefGoogle Scholar
  7. 7.
    Singh S, Singh DJ, Jaidka S, Somani R (2011) Comparative clinical evaluation of chemomechanical caries removal agent Papacarie® with conventional method among rural population in India: in vivo study. Braz J Oral Sci 10(3):193–198Google Scholar
  8. 8.
    Bussadori S, Guedes C, Bachiega J, Santis T, Motta L (2011) Clinical and radiographic study of chemical-mechanical removal of caries using Papacarie: 24-month follow up. J Clin Pediatr Dent 35(3):251–254CrossRefGoogle Scholar
  9. 9.
    Kotb RMS, Abdella AA, El Kateb MA, Ahmed AM (2009) Clinical evaluation of Papacarie in primary teeth. J Clin Pediatr Dent 34(2):117–123CrossRefGoogle Scholar
  10. 10.
    Aguirre Aguilar AA, Rios Caro TE, Huamán Saavedra J, França CM, Fernandes KPS, Mesquita-Ferrari RA et al (2012) Atraumatic restorative treatment: a dental alternative well-received by children. Rev Panam Salud Publica 31(2):148–152Google Scholar
  11. 11.
    Carrillo C, Tanaka M, Cesar M, Camargo M, Juliano Y, Novo N (2008) Use of papain gel in disabled patients. J Dent Child 75(3):222–228Google Scholar
  12. 12.
    Jawa D, Singh S, Somani R, Jaidka S, Sirkar K, Jaidka R (2010) Comparative evaluation of the efficacy of chemomechanical caries removal agent (Papacarie) and conventional method of caries removal: an in vitro study. J Indian Soc Pedod Prev Dent 28(2):73CrossRefGoogle Scholar
  13. 13.
    Matsumoto SFB, Motta LJ, Alfaya TA, Guedes CC, Fernandes KPS, Bussadori SK (2013) Assessment of chemomechanical removal of carious lesions using Papacarie Duo™: randomized longitudinal clinical trial. Indian J Dent Res 24(4):488CrossRefGoogle Scholar
  14. 14.
    Kush A, Thakur R, Patil SDS, Paul ST, Kakanur M (2015) Evaluation of antimicrobial action of Carie Care™ and Papacarie Duo™ on Aggregatibacter actinomycetemcomitans a major periodontal pathogen using polymerase chain reaction. Contemp Clin Dent 6(4):534CrossRefGoogle Scholar
  15. 15.
    Fleischer T, Ameade E, Mensah M, Sawer I (2003) Antimicrobial activity of the leaves and seeds of Bixa orellana. Fitoterapia. 74(1):136–138CrossRefGoogle Scholar
  16. 16.
    Shahid-ul-Islam, Rather LJ, Mohammad F (2016) Phytochemistry, biological activities and potential of annatto in natural colorant production for industrial applications – A review. Journal of Advanced Research 7 (3):499–514Google Scholar
  17. 17.
    Galindo-Cuspinera V, Rankin SA (2005) Bioautography and chemical characterization of antimicrobial compound (s) in commercial water-soluble annatto extracts. J Agric Food Chem 53(7):2524–2529CrossRefGoogle Scholar
  18. 18.
    Betsy J, Prasanth CS, Baiju KV, Prasanthila J, Subhash N (2014) Efficacy of antimicrobial photodynamic therapy in the management of chronic periodontitis: a randomized controlled clinical trial. J Clin Periodontol 41:573–581CrossRefGoogle Scholar
  19. 19.
    Deppe H, Mucke T, Wagenpfeil S, Kesting M, Sculean A (2013) Nonsurgical antimicrobial photodynamic therapy in moderate vs severe peri-implant defects: a clinical pilot study. Quintessence Int 44:609–618Google Scholar
  20. 20.
    Van Cuong T, Chin KB (2016) Effects of annatto (Bixa orellana L.) seeds powder on physicochemical properties, antioxidant and antimicrobial activities of pork patties during refrigerated storage. Korean J Food Sci Anim Resour 36(4):476CrossRefGoogle Scholar
  21. 21.
    Garcez AS, Neto JG, Sellera DP, Fregnani E (2015) Effects of antimicrobial photodynamic therapy and surgical endodontic treatment on the bacterial load reduction and periapical lesion healing. Three years follow up. Photodiagn Photodyn Ther 12:575–580CrossRefGoogle Scholar
  22. 22.
    Frencken JE, Makoni E, Sithole WD (1996) Atraumatic restorative treatment and glass ionomer cement sealants in school oral health programme in Zimbawe. Evaluation after 1 year. Caries Res 30(6):428–33.
  23. 23.
    Brostek A (2003) Early diagnosis and minimally invasive treatment of occlusal caries--a clinical approach. Oral Health Prev Dent 2:313–319Google Scholar
  24. 24.
    Whitaker EJ (2006) Primary, secondary and tertiary treatment of dental caries: a 20-year case report. J Am Dent Assoc 137(3):348–352CrossRefGoogle Scholar
  25. 25.
    Botta SB, Ana PA, Santos MO, Zezell DM, Matos AB (2012) Effect of dental tissue conditioners and matrix metalloproteinase inhibitors on type I collagen microstructure analyzed by Fourier transform infrared spectroscopy. J Biomed Mater Res B Appl Biomater 100(4):1009–1016CrossRefGoogle Scholar
  26. 26.
    Williams RW, Dunker AK (1981) Determination of the secondary structure of proteins from the amide I band of the laser Raman spectrum. J Mol Biol 152(4):783–813CrossRefGoogle Scholar
  27. 27.
    Susi H (1972) Infrared spectroscopy--conformation. Methods Enzymol 26:455CrossRefGoogle Scholar
  28. 28.
    Ramabrahmam V ( 2005) Being and becoming: a physics and Upanishadic awareness of time and thought process.Google Scholar
  29. 29.
    Sylvester M, Yannas I, Salzman E, Forbes M (1989) Collagen banded fibril structure and the collagen-platelet reaction. Thromb Res 55(1):135–148CrossRefGoogle Scholar
  30. 30.
    Stuart B (2005) Infrared spectroscopy: Wiley online library.
  31. 31.
    Karoui R, Downey G, Blecker C (2010) Mid-infrared spectroscopy coupled with chemometrics: a tool for the analysis of intact food systems and the exploration of their molecular structure− quality relationships− a review. Chem Rev 110(10):6144–6168CrossRefGoogle Scholar
  32. 32.
    Júnior ZSS, Botta SB, Ana PA, França CM, Fernandes KPS, Mesquita-Ferrari RA et al (2015) Effect of papain-based gel on type I collagen-spectroscopy applied for microstructural analysis. Sci Rep 23(5):11448.
  33. 33.
    George A, Veis A (1991) FTIRS in water demonstrates that collagen monomers undergo a conformational transition prior to thermal self-assembly in vitro. Biochemistry. 30(9):2372–2377CrossRefGoogle Scholar
  34. 34.
    Rojas JC, Bruchey AK, Gonzalez-Lima F (2012) Neurometabolic mechanisms for memory enhancement and neuroprotection of methylene blue. Prog Neurobiol 96(1):32–45CrossRefGoogle Scholar
  35. 35.
    Ericson D, Kidd E, McComb D, Mjör I, Noack MJ (2003) Minimally invasive dentistry--concepts and techniques in cariology. Oral Health Prev Dent 1:1Google Scholar
  36. 36.
    Bittencourt S, Pereira J, Rosa A, Oliveira K, Ghizoni J, Oliveira M (2010) Mineral content removal after Papacarie application in primary teeth: a quantitative analysis. J Clin Pediatr Dent 34(3):229–231CrossRefGoogle Scholar
  37. 37.
    Banerjee A, Frencken JE, Schwendicke F3, Innes NPT (2017) Contemporary operative caries management: consensus recommendations on minimally invasivecaries removal. Br Dent J 223(3):215–222.
  38. 38.
    Mackenzie L, Banerjee A (2017) Minimally invasive direct restorations: a practical guide. Br Dent J 223(3):163–171.

Copyright information

© Springer-Verlag London Ltd., part of Springer Nature 2019

Authors and Affiliations

  • Zenildo Santos SilvaJr
    • 1
    • 2
    • 3
    • 4
    Email author
  • Cristiane Miranda França
    • 3
    • 4
  • Renato Araújo Prates
    • 3
    • 4
  • Sergio Brossi Botta
    • 3
    • 4
  • Raquel Agnelli Mesquita Ferrari
    • 3
  • Patricia Aparecida Ana
    • 5
  • Christiane Pavani
    • 3
  • Kristianne Porta Santos Fernandes
    • 3
    • 4
  • Daniela de Fátima Teixeira da Silva
    • 3
  • Michael R. Hamblin
    • 1
    • 2
    • 6
  • Sandra Kalil Bussadori
    • 3
    • 4
    Email author
  1. 1.Massachusetts General HospitalWellman Center for PhotomedicineBostonUSA
  2. 2.Department of DermatologyHarvard Medical SchoolBostonUSA
  3. 3.Postgraduate Program in Biophotonics Applied to Health SciencesNove de Julho UniversitySão PauloBrazil
  4. 4.School of DentistryNove de Julho UniversitySão PauloBrazil
  5. 5.Center of Engineering, Modeling and Applied Social SciencesFederal University of ABCSão Bernardo do CampoBrazil
  6. 6.Harvard-MITDivision of Health Sciences and TechnologyCambridgeUSA

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