Clinical Oral Investigations

, Volume 20, Issue 8, pp 2315–2320 | Cite as

Stain removal effect of novel papain- and bromelain-containing gels applied to enamel

  • Eliseu A. Münchow
  • Henry J. Hamann
  • M. Teresa Carvajal
  • Rodolfo PinalEmail author
  • Marco C. BottinoEmail author
Short Communication



The aims of the study are to prepare novel stain removal gel-based formulations containing papain or bromelain and to investigate their stain removal effect when applied to enamel.

Materials and methods

Experimental bromelain- and papain-based stain removal gels were prepared. Next, enamel/dentin tooth samples (6 × 6 mm2, 4 mm in thickness) were obtained from bovine teeth, stained in coffee solution for 1 week, and measured with a digital spectrophotometer (Easyshade, Vita Zahnfabrik) for color assessment (baseline). The samples were then randomly allocated into four groups (n = 7), according to the stain removal agent applied: ContrastPM+ (Discus Dental, LLC), which is based on 20 wt.% carbamide peroxide (positive control); bromelain-based; papain-based; and no agent (negative control). The materials were applied once a week, three times per day, during 4 weeks, and following the directions of use from positive control. The samples were measured again with the Easyshade and using the CIEL * a * b * color system. The color change (ΔE *) results were obtained by subtracting the baseline values from the final color values obtained at each time point. The data were statistically analyzed using two-way repeated-measures analysis of variance and Student Newman Keuls’s test as a post hoc test (α = 5 %).


All stain removal agents produced greater color change than the negative control (p < .001), with the positive control demonstrating greater ΔE * values when compared to the experimental gels (p ≤ .004). The second application of all gels resulted in greater ΔE * values compared to the first application (p ≤ .025), although no color change was observed after the third application (p ≥ .051), regardless of the material evaluated.

Clinical significance

The proposed gels containing proteolytic enzymes (bromelain or papain) of vegetal origin may hold significant clinical potential as active agents for the preparation of stain removal agents free of hydrogen/carbamide peroxide.


Extrinsic stain removal Carbamide peroxide Proteolytic enzymes Digital spectrophotometer 



The authors thank Mr. Cody Spoolstra—Purdue Student Soybean Product Innovation Competition through the fund of Indiana Soybean Alliance.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.


This study was funded in part by start-up funds from the IU School of Dentistry (M.C.B.).

Ethical approval

This article does not contain any studies with human participants or animals performed by any of the authors.

Informed consent

Not applicable.


  1. 1.
    Haywood VB, Heymann HO (1989) Nightguard vital bleaching. Quintessence Int 20:173–176PubMedGoogle Scholar
  2. 2.
    Giachetti L, Bertini F, Bambi C, Nieri M, Scaminaci Russo D (2010) A randomized clinical trial comparing at-home and in-office tooth whitening techniques: a nine-month follow-up. J Am Dent Assoc 141:1357–1364CrossRefPubMedGoogle Scholar
  3. 3.
    Kielbassa AM, Maier M, Gieren AK, Eliav E (2015) Tooth sensitivity during and after vital tooth bleaching: a systematic review on an unsolved problem. Quintessence Int 46:881–897PubMedGoogle Scholar
  4. 4.
    de Paula EA, Nava JA, Rosso C, Benazzi CM, Fernandes KT, Kossatz S, Loguercio AD, Reis A (2014) In-office bleaching with a two- and seven-day intervals between clinical sessions: a randomized clinical trial on tooth sensitivity. J Dent 43:424–429. doi: 10.1016/j.jdent.2014.09.009 CrossRefPubMedGoogle Scholar
  5. 5.
    Almeida LC, Riehl H, Santos PH, Sundfeld ML, Briso AL (2012) Clinical evaluation of the effectiveness of different bleaching therapies in vital teeth. Int J Periodontics Restorative Dent 32:303–309PubMedGoogle Scholar
  6. 6.
    He LB, Shao MY, Tan K, Xu X, Li JY (2012) The effects of light on bleaching and tooth sensitivity during in-office vital bleaching: a systematic review and meta-analysis. J Dent 40:644–653. doi: 10.1016/j.jdent.2012.04.010 CrossRefPubMedGoogle Scholar
  7. 7.
    Reis A, Dalanhol AP, Cunha TS, Kossatz S, Loguercio AD (2011) Assessment of tooth sensitivity using a desensitizer before light-activated bleaching. Oper Dent 36:12–17. doi: 10.2341/10-148-CR CrossRefPubMedGoogle Scholar
  8. 8.
    Duque CC, Soares DG, Basso FG, Hebling J, de Souza Costa CA (2014) Bleaching effectiveness, hydrogen peroxide diffusion, and cytotoxicity of a chemically activated bleaching gel. Clin Oral Investig 18:1631–1637. doi: 10.1007/s00784-013-1147-4 PubMedGoogle Scholar
  9. 9.
    Soares DG, Ribeiro AP, da Silveira VF, Hebling J, de Souza Costa CA (2013) Efficacy and cytotoxicity of a bleaching gel after short application times on dental enamel. Clin Oral Investig 17:1901–1909. doi: 10.1007/s00784-012-0883-1 CrossRefPubMedGoogle Scholar
  10. 10.
    Mena-Serrano AP, Parreiras SO, do Nascimento EM, Borges CP, Berger SB, Loguercio AD and Reis A (2015) Effects of the concentration and composition of in-office bleaching gels on hydrogen peroxide penetration into the pulp chamber. Oper Dent 40:E76–E82. doi: 10.2341/13-352-L
  11. 11.
    Bolay S, Cakir FY, Gurgan S (2012) Effects of toothbrushing with fluoride abrasive and whitening dentifrices on both unbleached and bleached human enamel surface in terms of roughness and hardness: an in vitro study. J Contemp Dent Pract 13:584–589PubMedGoogle Scholar
  12. 12.
    Ernst CP, Marroquin BB, Willershausen-Zonnchen B (1996) Effects of hydrogen peroxide-containing bleaching agents on the morphology of human enamel. Quintessence Int 27:53–56PubMedGoogle Scholar
  13. 13.
    Hegedus C, Bistey T, Flora-Nagy E, Keszthelyi G, Jenei A (1999) An atomic force microscopy study on the effect of bleaching agents on enamel surface. J Dent 27:509–515CrossRefPubMedGoogle Scholar
  14. 14.
    Faraoni-Romano JJ, Da Silveira AG, Turssi CP, Serra MC (2008) Bleaching agents with varying concentrations of carbamide and/or hydrogen peroxides: effect on dental microhardness and roughness. J Esthet Restor Dent 20:395–402. doi: 10.1111/j.1708-8240.2008.00216.x CrossRefPubMedGoogle Scholar
  15. 15.
    Caviedes-Bucheli J, Ariza-Garcia G, Restrepo-Mendez S, Rios-Osorio N, Lombana N, Munoz HR (2008) The effect of tooth bleaching on substance P expression in human dental pulp. J Endod 34:1462–1465. doi: 10.1016/j.joen.2008.09.013 CrossRefPubMedGoogle Scholar
  16. 16.
    Gopinath S, James V, Vidhya S, Karthikeyan K, Kavitha S, Mahalaxmi S (2013) Effect of bleaching with two different concentrations of hydrogen peroxide containing sweet potato extract as an additive on human enamel: an in vitro spectrophotometric and scanning electron microscopy analysis. J Conserv Dent 16:45–49. doi: 10.4103/0972-0707.105298 CrossRefPubMedPubMedCentralGoogle Scholar
  17. 17.
    Kalyana P, Shashidhar A, Meghashyam B, Sreevidya KR, Sweta S (2011) Stain removal efficacy of a novel dentifrice containing papain and Bromelain extracts—an in vitro study. Int J Dent Hyg 9:229–233. doi: 10.1111/j.1601-5037.2010.00473.x CrossRefPubMedGoogle Scholar
  18. 18.
    Chakravarthy P, Acharya S (2012) Efficacy of extrinsic stain removal by novel dentifrice containing papain and bromelain extracts. J Young Pharm 4:245–249. doi: 10.4103/0975-1483.104368 CrossRefPubMedPubMedCentralGoogle Scholar
  19. 19.
    Meireles SS, Fontes ST, Coimbra LA, Della Bona A, Demarco FF (2012) Effectiveness of different carbamide peroxide concentrations used for tooth bleaching: an in vitro study. J Appl Oral Sci 20:186–191CrossRefPubMedPubMedCentralGoogle Scholar
  20. 20.
    Commission Internationale De L’Eclairage (1978) Recommendations on uniform colour spaces, colour difference equations and psychometric colour terms. Book title, Paris: Bureau Central de la CIEGoogle Scholar
  21. 21.
    Plotino G, Buono L, Grande NM, Pameijer CH, Somma F (2008) Nonvital tooth bleaching: a review of the literature and clinical procedures. J Endod 34:394–407. doi: 10.1016/j.joen.2007.12.020 CrossRefPubMedGoogle Scholar
  22. 22.
    Sabatini C, Campillo M, Aref J (2012) Color stability of ten resin-based restorative materials. J Esthet Restor Dent 24:185–199. doi: 10.1111/j.1708-8240.2011.00442.x CrossRefPubMedGoogle Scholar
  23. 23.
    Sato C, Rodrigues FA, Garcia DM, Vidal CM, Pashley DH, Tjaderhane L, Carrilho MR, Nascimento FD, Tersariol IL (2013) Tooth bleaching increases dentinal protease activity. J Dent Res 92:187–192. doi: 10.1177/0022034512470831 CrossRefPubMedGoogle Scholar
  24. 24.
    de Silva GM, Brackett MG, Haywood VB (2006) Number of in-office light-activated bleaching treatments needed to achieve patient satisfaction. Quintessence Int 37:115–120Google Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2016

Authors and Affiliations

  1. 1.Department of Biomedical and Applied Sciences, Division of Dental BiomaterialsIndiana University School of Dentistry (IUSD)IndianapolisUSA
  2. 2.Graduate Program in Dentistry, School of DentistryFederal University of Pelotas–UFPelPelotasBrazil
  3. 3.Department of Industrial and Physical Pharmacy, College of PharmacyPurdue UniversityWest LafayetteUSA
  4. 4.Department of Agricultural and Biological EngineeringPurdue UniversityWest LafayetteUSA

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