Restorative Dentistry and Endodontics

The Korean Academy of Conservative Dentistry (대한치과보존학회)
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Antifungal effects of synthetic human β-defensin 3-C15 peptide

  • Lim, Sang-Min (Department of Conservative Dentistry, Seoul National University Dental Hospital, Seoul National University School of Dentistry and Dental Research Institute) ;
  • Ahn, Ki-Bum (Department of Oral Microbiology and Immunology, DRI and BK21 Plus Program, Seoul National University School of Dentistry) ;
  • Kim, Christine (Department of Anatomy and Cell Biology, McGill University) ;
  • Kum, Jong-Won (Department of Biology, Siena College) ;
  • Perinpanayagam, Hiran (Schulich School of Medicine and Dentistry, University of Western Ontario) ;
  • Gu, Yu (Department of Conservative Dentistry, Seoul National University Dental Hospital, Seoul National University School of Dentistry and Dental Research Institute) ;
  • Yoo, Yeon-Jee (Department of Conservative Dentistry, Seoul National University Dental Hospital, Seoul National University School of Dentistry and Dental Research Institute) ;
  • Chang, Seok Woo (Department of Conservative Dentistry, School of Dentistry, Kyung Hee University) ;
  • Han, Seung Hyun (Department of Oral Microbiology and Immunology, DRI and BK21 Plus Program, Seoul National University School of Dentistry) ;
  • Shon, Won-Jun (Department of Conservative Dentistry, Seoul National University Dental Hospital, Seoul National University School of Dentistry and Dental Research Institute) ;
  • Lee, Woocheol (Department of Conservative Dentistry, Seoul National University Dental Hospital, Seoul National University School of Dentistry and Dental Research Institute) ;
  • Baek, Seung-Ho (Department of Conservative Dentistry, Seoul National University Dental Hospital, Seoul National University School of Dentistry and Dental Research Institute) ;
  • Zhu, Qiang (Division of Endodontology, Department of Oral Health and Diagnostic Sciences, University of Connecticut Health Center, School of Dental Medicine) ;
  • Kum, Kee-Yeon (Department of Conservative Dentistry, Seoul National University Dental Hospital, Seoul National University School of Dentistry and Dental Research Institute)
  • Received : 2015.10.23
  • Accepted : 2016.02.16
  • Published : 2016.05.31
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Abstract

Objectives: The purpose of this ex vivo study was to compare the antifungal activity of a synthetic peptide consisting of 15 amino acids at the C-terminus of human ${\beta}$-defensin 3 (HBD3-C15) with calcium hydroxide (CH) and Nystatin (Nys) against Candida albicans (C. albicans) biofilm. Materials and Methods: C. albicans were grown on cover glass bottom dishes or human dentin disks for 48 hr, and then treated with HBD3-C15 (0, 12.5, 25, 50, 100, 150, 200, and $300{\mu}g/mL$), CH ($100{\mu}g/mL$), and Nys ($20{\mu}g/mL$) for 7 days at $37^{\circ}C$. On cover glass, live and dead cells in the biomass were measured by the FilmTracer Biofilm viability assay, and observed by confocal laser scanning microscopy (CLSM). On dentin, normal, diminished and ruptured cells were observed by field-emission scanning electron microscopy (FE-SEM). The results were subjected to a two-tailed t-test, a one way analysis variance and a post hoc test at a significance level of p = 0.05. Results: C. albicans survival on dentin was inhibited by HBD3-C15 in a dose-dependent manner. There were fewer aggregations of C. albicans in the groups of Nys and HBD3-C15 (${\geq}100{\mu}g/mL$). CLSM showed C. albicans survival was reduced by HBD3-C15 in a dose dependent manner. Nys and HBD3-C15 (${\geq}100{\mu}g/mL$) showed significant fungicidal activity compared to CH group (p < 0.05). Conclusions: Synthetic HBD3-C15 peptide (${\geq}100{\mu}g/mL$) and Nys exhibited significantly higher antifungal activity than CH against C. albicans by inhibiting cell survival and biofilm.

Keywords

References

  1. Waltimo TMT, Haapasalo M, Zehnder M, Meyer J. Clinical aspects related to endodontic yeast infections. Endod Topics 2004;9:66-78. https://doi.org/10.1111/j.1601-1546.2004.00086.x
  2. Naglik JR, Rodgers CA, Shirlaw PJ, Dobbie JL, Fernandes-Naglik LL, Greenspan D, Agabian N, Challacombe SJ. Differential expression of Candida albicans secreted aspartyl proteinase and phospholipase B genes in humans correlates with active oral and vaginal infections. J Infect Dis 2003;188:469-479. https://doi.org/10.1086/376536
  3. Siqueira JF Jr, Rocas IN, Lopes HP, Elias CN, de Uzeda M. Fungal infection of the radicular dentin. J Endod 2002;28:770-773. https://doi.org/10.1097/00004770-200211000-00006
  4. Mitchell KF, Zarnowski R, Sancheza H, Edward JA, Reinicke EL, Nett JE, Mitchell AP, Andes DR. Community participation in biofilm matrix assembly and function. Proc Natl Acad Sci U S A 2015;112:4092-4097. https://doi.org/10.1073/pnas.1421437112
  5. Kumamoto CA. Candida biofilms. Curr Opin Microbiol 2002:5:608-611. https://doi.org/10.1016/S1369-5274(02)00371-5
  6. Mah TF, O'Toole GA. Mechanisms of biofilm resistance to antimicrobial agents. Trends Microbiol 2001;9:34-39. https://doi.org/10.1016/S0966-842X(00)01913-2
  7. Sundqvist G, Figdor D, Persson S, Sjogren U. Microbiologic analysis of teeth with failed endodontic treatment and the outcome of conservative retreatment. Oral Surg Oral Med Oral Pathol Oral Radiol Endod 1998;85:86-93. https://doi.org/10.1016/S1079-2104(98)90404-8
  8. Nair PN, Sjogren U, Krey G, Kahnberg KE, Sundqvist G. Intraradicular bacteria and fungi in root-filled, asymptomatic human teeth with therapy-resistant periapical Lesions: a long-term light and electron microscopic follow-up study. J Endod 1990;16:580-588. https://doi.org/10.1016/S0099-2399(07)80201-9
  9. Bystrom A, Sundqvist G. Bacteriologic evaluation of the efficacy of mechanical root canal instrumentation in endodontic therapy. Scand J Dent Res 1981;89:321-328.
  10. Gupta SP, Bhati M, Jhajharia K, Patel H, Paliwal A, Franklin S. Evaluation of antimicrobial and antifungal efficacy of inter appointment intracanal medicaments against Enterococcus and Candida albicans: an in vitro study. J Int Oral Health 2015;7:97-102.
  11. Law A, Messer H. An evidence-based analysis of the antibacterial effectiveness of intracanal medicaments. J Endod 2004;30:689-694. https://doi.org/10.1097/01.DON.0000129959.20011.EE
  12. Siqueira JF Jr, Sen BH. Fungi in endodontic infections. Oral Surg Oral Med Oral Pathol Oral Radiol Endod 2004;97:632-641. https://doi.org/10.1016/j.tripleo.2003.12.022
  13. Rosato A, Vitali C, Piarulli M, Mazzotta M, Argentieri MP, Mallamaci R. In vitro synergic efficacy of the combination of Nystatin with the essential oils of Origanum vulgare and Pelargonium graveolens against some Candida species. Phytomedicine 2009;16:972-975. https://doi.org/10.1016/j.phymed.2009.02.011
  14. Kaomongkolgit R, Jamdee K, Chaisomboon N. Antifungal activity of alpha-mangostin against Candida albicans. J Oral Sci 2009;51:401-406. https://doi.org/10.2334/josnusd.51.401
  15. Lima SM, de Padua GM, Sousa MG, Freire Mde S, Franco OL, Rezende TM. Antimicrobial peptide-based treatment for endodontic infections-biotechnological innovation in endodontics. Biotechnol Adv 2015;33:203-213. https://doi.org/10.1016/j.biotechadv.2014.10.013
  16. Paris S, Wolgin M, Kielbassa AM, Pries A, Zakrzewicz A. Gene expression of human beta-defensins in healthy and inflamed human dental pulps. J Endod 2009;35:520-523. https://doi.org/10.1016/j.joen.2008.12.015
  17. Schibli DJ, Hunter HN, Aseyev V, Starner TD, Wiencek JM, McCray PB Jr, Tack BF, Vogel HJ. The solution structures of the human beta-defensins lead to a better understanding of the potent bactericidal activity of HBD3 against Staphylococcus aureus. J Biol Chem 2002;277:8279-8289. https://doi.org/10.1074/jbc.M108830200
  18. Dhople V, Krukemeyer A, Ramamoorthy A. The human beta-defensin-3, an antibacterial peptide with multiple biological functions. Biochim Biophys Acta 2006;1758:1499-1512. https://doi.org/10.1016/j.bbamem.2006.07.007
  19. Lee JK, Park YJ, Kum KY, Han SH, Chang SW, Kaufman B, Jiang J, Zhu Q, Safavi K, Spangberg L. Antimicrobial efficacy of a human beta-defensin-3 peptide using an Enterococcus faecalis dentine infection model. Int Endod J 2013;46:406-412. https://doi.org/10.1111/iej.12002
  20. Lee JK, Chang SW, Perinpanayagam H, Lim SM, Park YJ, Han SH, Baek SH, Zhu Q, Bae KS, Kum KY. Antibacterial efficacy of a human beta-defensin-3 peptide on multispecies biofilms. J Endod 2013;39:1625-1629. https://doi.org/10.1016/j.joen.2013.07.035
  21. Krishnakumari V, Rangaraj N, Nagaraj R. Antifungal activities of human beta-defensins HBD-1 to HBD-3 and their C-terminal analogs Phd1 to Phd3. Antimicrob Agents Chemother 2009;53:256-260. https://doi.org/10.1128/AAC.00470-08
  22. Hoover DM, Wu Z, Tucker K, Lu W, Lubkowski J. Antimicrobial characterization of human beta-defensin 3 derivatives. Antimicrob Agents Chemother 2003;47:2804-2809. https://doi.org/10.1128/AAC.47.9.2804-2809.2003
  23. Song W, Shi Y, Xiao M, Lu H, Qu T, Li P, Wu G, Tian Y. In vitro bactericidal activity of recombinant human beta-defensin-3 against pathogenic bacterial strains in human tooth root canal. Int J Antimicrob Agents 2009;33:237-243. https://doi.org/10.1016/j.ijantimicag.2008.05.022
  24. Taff HT, Nett JE, Andes DR. Comparative analysis of Candida biofilm quantitation assays. Med Mycol 2012;50:214-218. https://doi.org/10.3109/13693786.2011.580016
  25. Guerreiro-Tanomaru JM, de Faria-Junior NB, Duarte MA, Ordinola-Zapata R, Graeff MS, Tanomaru-Filho M. Comparative analysis of Enterococcus faecalis biofilm formation on different substrates. J Endod 2013;39:346-350. https://doi.org/10.1016/j.joen.2012.09.027
  26. Kim D, Kim E. Antimicrobial effect of calcium hydroxide as an intracanal medicament in root canal treatment: a literature review - Part II. in vivo studies. Restor Dent Endod 2015;40:97-103. https://doi.org/10.5395/rde.2015.40.2.97
  27. Mohammadi Z, Shalavi S, Yazdizadeh M. Antimicrobial activity of calcium hydroxide in endodontics: a review. Chonnam Med J 2012;48:133-140. https://doi.org/10.4068/cmj.2012.48.3.133
  28. Wang JD, Hume WR. Diffusion of hydrogen ion and hydroxyl ion from various sources through dentine. Int Endod J 1988;21:17-26. https://doi.org/10.1111/j.1365-2591.1988.tb00949.x
  29. Kinsky SC, Nystatin binding by protoplasts and a particulate fraction of Neurospora crassa, and a basis for the selective toxicity of polyene antifungal antibiotics. Proc Natl Acad Sci U S A 1962;48:1049-1056. https://doi.org/10.1073/pnas.48.6.1049
  30. de Kruijff BD, Demel RA. Polyene antibiotic-sterol interactions in membranes of Acholeplasma laidlawii cells and lecithin liposomes. III. Molecular structure of the polyene antibiotic-cholesterol complexes. Biochim Biophys Acta 1974;339:57-70. https://doi.org/10.1016/0005-2736(74)90332-0
  31. Mohammadi Z. Systemic, prophylactic and local applications of antimicrobials in endodontics: an update review. Int Dent J 2009;59:175-186.
  32. Vylkova S, Li XS, Berner JC, Edgerton M. Distinct antifungal mechanisms: beta-defensins require Candida albicans Ssa1 protein, while Trk1p mediates activity of cysteine-free cationic peptides. Antimicrob Agents Chemother 2006;50:324-331. https://doi.org/10.1128/AAC.50.1.324-331.2006
  33. Feng Z, Jiang B, Chandra J, Ghannoum M, Nelson S, Weinberg A. Human beta-defensins: differential activity against Candidal species and regulation by Candida albicans. J Dent Res 2005;84:445-450. https://doi.org/10.1177/154405910508400509
  34. Kluver E, Schulz-Maronde S, Scheid S, Meyer B, Forssmann WG, Adermann K. Structure-activity relation of human $\beta$-defensin 3: influence of disulfide bonds and cysteine substitution on antimicrobial activity and cytotoxicity. Biochemistry 2005;44:9804-9816. https://doi.org/10.1021/bi050272k
  35. Maisetta G, Batoni G, Esin S, Luperini F, Pardini M, Bottai D, Florio W, Giuca MR, Gabriele M, Campa M. Activity of human $\beta$-defensin 3 alone or combined with other antimicrobial agents against oral bacteria. Antimicrob Agents Chemother 2003;47:3349-3351. https://doi.org/10.1128/AAC.47.10.3349-3351.2003

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