Skip to main content

Advertisement

Log in

Effect of ozone on periodontopathogenic species—an in vitro study

  • Original Article
  • Published:
Clinical Oral Investigations Aims and scope Submit manuscript

Abstract

The in vitro study was aimed to determine the effect of ozone on periodontopathogenic microorganisms. Ozone was generated for 6 s–2 × 24 s (corresponding to 0.56 mg–2 × 2.24 mg of ozone) against 23 mainly anaerobic periodontopathogenic species. Agar diffusion test was used as a screening method. Then, the killing activity was tested in a serum-free environment and with 25% v/v inactivated serum. Further, the effect of ozone on bactericidal activity of native serum was analyzed against Fusobacterium nucleatum, Porphyromonas gingivalis, and Aggregatibacter actinomycetemcomitans. Agar diffusion test showed a high efficacy of ozone against microorganisms, especially against Porphyromonas gingivalis. This result was confirmed by the killing tests; most of the strains in a concentration of 105 were completely eliminated after twofold 18-s application of ozone. Only four of the six potentially “superinfecting” species (Staphylococcus aureus, Enterococcus faecalis, Enterobacter cloacae, Candida albicans) survived in part. Addition of heat-inactivated serum reduced the killing rate of ozone by 78% after 6-s and by 47% after twofold 18-s exposures; no strain was completely eradicated after any application of ozone. The bactericidal effect of native serum was enhanced after application of ozone; no effect was visible on the included A. actinomycetemcomitans strain which was found to be completely resistant to the bactericidal action of serum. In conclusion, (a) ozone has a strong antibacterial activity against putative periodontopathogenic microorganisms, and (b) the bactericidal effect is reduced in the presence of serum. Ozone may have potential as an adjunctive application to mechanical treatment in periodontitis patients.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  1. Socransky SS, Haffajee AD (2005) Periodontal microbial ecology. Periodontol 2000 38:135–187

    Google Scholar 

  2. Consensus report (1996) Periodontal diseases: pathogenesis and microbial factors. Ann Periodontol 1:926–932

    Google Scholar 

  3. Socransky SS, Haffajee AD, Cugini MA, Smith C, Kent RL Jr (1998) Microbial complexes in subgingival plaque. J Clin Periodontol 25:134–144

    Article  PubMed  Google Scholar 

  4. Hultin M, Gustafsson A, Hallstrom H, Johansson LA, Ekfeldt A, Klinge B (2002) Microbiological findings and host response in patients with peri-implantitis. Clin Oral Implants Res 13:349–358

    Article  PubMed  Google Scholar 

  5. Van de Velde T, Thevissen E, Persson GR, Johansson C, De Bruyn H (2009) Two-year outcome with Nobel Direct implants: a retrospective radiographic and microbiologic study in 10 patients. Clin Implant Dent Relat Res 11:183–193

    Article  PubMed  Google Scholar 

  6. Schenkein HA, Genco RJ (1977) Gingival fluid and serum in periodontal diseases. I. Quantitative study of immunoglobulins, complement components, and other plasma proteins. J Periodontol 48:772–777

    PubMed  Google Scholar 

  7. Selk SH, Pogany SA, Higuchi T (1982) Comparative antimicrobial activity, in vitro and in vivo, of soft N-chloramine systems and chlorhexidine. Appl Environ Microbiol 43:899–904

    PubMed  Google Scholar 

  8. Santos S, Herrera D, Lopez E, O'Connor A, Gonzalez I, Sanz M (2004) A randomized clinical trial on the short-term clinical and microbiological effects of the adjunctive use of a 0.05% chlorhexidine mouth rinse for patients in supportive periodontal care. J Clin Periodontol 31:45–51

    Article  PubMed  Google Scholar 

  9. Cosyn J, Sabzevar MM (2007) Subgingival chlorhexidine varnish administration as an adjunct to same-day full-mouth root planing. II. Microbiological observations. J Periodontol 78:438–445

    Article  PubMed  Google Scholar 

  10. van Winkelhoff AJ, Winkel EG (2009) Antibiotics in periodontics: right or wrong? J Periodontol 80:1555–1558

    Article  PubMed  Google Scholar 

  11. Guentsch A, Jentsch H, Pfister W, Hoffmann T, Eick S (2008) Moxifloxacin as an adjunctive antibiotic in the treatment of severe chronic periodontitis. J Periodontol 79:1894–1903

    Article  PubMed  Google Scholar 

  12. Tomasi C, Schander K, Dahlen G, Wennstrom JL (2006) Short-term clinical and microbiologic effects of pocket debridement with an Er:YAG laser during periodontal maintenance. J Periodontol 77:111–118

    Article  PubMed  Google Scholar 

  13. Das E, Gurakan GC, Bayindirli A (2006) Effect of controlled atmosphere storage, modified atmosphere packaging and gaseous ozone treatment on the survival of Salmonella Enteritidis on cherry tomatoes. Food Microbiol 23:430–438

    Article  PubMed  Google Scholar 

  14. Murakami H, Mizuguchi M, Hattori M, Ito Y, Kawai T, Hasegawa J (2002) Effect of denture cleaner using ozone against methicillin-resistant Staphylococcus aureus and E. coli T1 phage. Dent Mater J 21:53–60

    Article  PubMed  Google Scholar 

  15. Dyas A, Boughton BJ, Das BC (1983) Ozone killing action against bacterial and fungal species; microbiological testing of a domestic ozone generator. J Clin Pathol 36:1102–1104

    Article  PubMed  Google Scholar 

  16. Doroszkiewicz W, Sikorska I, Jankowski S (1993) Ozone as sensitizer of bacteria to the bactericidal action of complement. Acta Microbiol Pol 42:315–319

    PubMed  Google Scholar 

  17. Azarpazhooh A, Limeback H (2008) The application of ozone in dentistry: a systematic review of literature. J Dent 36:104–116

    Article  PubMed  Google Scholar 

  18. Nagayoshi M, Fukuizumi T, Kitamura C, Yano J, Terashita M, Nishihara T (2004) Efficacy of ozone on survival and permeability of oral microorganisms. Oral Microbiol Immunol 19:240–246

    Article  PubMed  Google Scholar 

  19. Feres M, Haffajee AD, Allard K, Som S, Goodson JM, Socransky SS (2002) Antibiotic resistance of subgingival species during and after antibiotic therapy. J Clin Periodontol 29:724–735

    Article  PubMed  Google Scholar 

  20. Paoloni M, Di Sante L, Cacchio A, Apuzzo D, Marotta S, Razzano M, Franzini M, Santilli V (2009) Intramuscular oxygen-ozone therapy in the treatment of acute back pain with lumbar disc herniation: a multicenter, randomized, double-blind, clinical trial of active and simulated lumbar paravertebral injection. Spine (Phila Pa 1976) 34:1337–1344

    Article  Google Scholar 

  21. Dharap SB, Ghag GS, Kulkarni KP, Venkatesh V (2008) Efficacy and safety of oxum in treatment of the venous ulcer. J Indian Med Assoc 106(326):328–330

    Google Scholar 

  22. Lenes D, Deboosere N, Menard-Szczebara F, Jossent J, Alexandre V, Machinal C, Vialette M (2010) Assessment of the removal and inactivation of influenza viruses H5N1 and H1N1 by drinking water treatment. Water Res 44:2473–2486

    Article  PubMed  Google Scholar 

  23. Choi Y, Cho M, Lee Y, Choi J, Yoon J (2007) Inactivation of Bacillus subtilis spores during ozonation in water treatment plant: influence of pre-treatment and consequences for positioning of the ozonation step. Chemosphere 69:675–681

    Article  PubMed  Google Scholar 

  24. Smeets PW, van der Helm AW, Dullemont YJ, Rietveld LC, van Dijk JC, Medema GJ (2006) Inactivation of Escherichia coli by ozone under bench-scale plug flow and full-scale hydraulic conditions. Water Res 40:3239–3248

    Article  PubMed  Google Scholar 

  25. Schwan L, Bamfaste M (1951) Experiences with the use of chlorine gas and ozone in the treatment of root gangrene and dental granuloma. Dtsch Zahnärztl Z 6:248–263

    PubMed  Google Scholar 

  26. Kronenberg O, Lussi A, Ruf S (2009) Preventive effect of ozone on the development of white spot lesions during multibracket appliance therapy. Angle Orthod 79:64–69

    Article  PubMed  Google Scholar 

  27. Baysan A, Lynch E (2007) Clinical reversal of root caries using ozone: 6-month results. Am J Dent 20:203–208

    PubMed  Google Scholar 

  28. Huth KC, Paschos E, Brand K, Hickel R (2005) Effect of ozone on non-cavitated fissure carious lesions in permanent molars. A controlled prospective clinical study. Am J Dent 18:223–228

    PubMed  Google Scholar 

  29. Huth KC, Quirling M, Maier S, Kamereck K, Alkhayer M, Paschos E, Welsch U, Miethke T, Brand K, Hickel R (2009) Effectiveness of ozone against endodontopathogenic microorganisms in a root canal biofilm model. Int Endod J 42:3–13

    Article  PubMed  Google Scholar 

  30. Yamada K, Yama M, Takaku Y, Kakizawa T, Kimizuka R, Okuda K, Kato T (2010) Antimicrobial activity of super-oxidised water against oral microorganisms. Arch Oral Biol 55:397–400

    Article  PubMed  Google Scholar 

  31. Sharma M, Hudson JB (2008) Ozone gas is an effective and practical antibacterial agent. Am J Infect Control 36:559–563

    Article  PubMed  Google Scholar 

  32. Hems RS, Gulabivala K, Ng YL, Ready D, Spratt DA (2005) An in vitro evaluation of the ability of ozone to kill a strain of Enterococcus faecalis. Int Endod J 38:22–29

    Article  PubMed  Google Scholar 

  33. Ozbek SM, Ozbek A, Erdorgan AS (2009) Analysis of Enterococcus faecalis in samples from Turkish patients with primary endodontic infections and failed endodontic treatment by real-time PCR SYBR green method. J Appl Oral Sci 17:370–374

    Article  PubMed  Google Scholar 

  34. Kustarci A, Sumer Z, Altunbas D, Kosum S (2009) Bactericidal effect of KTP laser irradiation against Enterococcus faecalis compared with gaseous ozone: an ex vivo study. Oral Surg Oral Med Oral Pathol Oral Radiol Endod 107:e73–e79

    Article  PubMed  Google Scholar 

  35. Tew JG, Marshall DR, Burmeister JA, Ranney RR (1985) Relationship between gingival crevicular fluid and serum antibody titers in young adults with generalized and localized periodontitis. Infect Immun 49:487–493

    PubMed  Google Scholar 

  36. Zeitlinger MA, Sauermann R, Traunmuller F, Georgopoulos A, Muller M, Joukhadar C (2004) Impact of plasma protein binding on antimicrobial activity using time-killing curves. J Antimicrob Chemother 54:876–880

    Article  PubMed  Google Scholar 

  37. Kawamura-Sato K, Wachino J, Kondo T, Ito H, Arakawa Y (2008) Reduction of disinfectant bactericidal activities in clinically isolated Acinetobacter species in the presence of organic material. J Antimicrob Chemother 61:568–576

    Article  PubMed  Google Scholar 

  38. Grenier D, Imbeault S, Plamondon P, Grenier G, Nakayama K, Mayrand D (2001) Role of gingipains in growth of Porphyromonas gingivalis in the presence of human serum albumin. Infect Immun 69:5166–5172

    Article  PubMed  Google Scholar 

  39. Burgassi S, Zanardi I, Travagli V, Montomoli E, Bocci V (2009) How much ozone bactericidal activity is compromised by plasma components? J Appl Microbiol 106:1715–1721

    Article  PubMed  Google Scholar 

  40. Kuboniwa M, Lamont RJ (2010) Subgingival biofilm formation. Periodontol 2000 52:38–52

    Article  PubMed  Google Scholar 

  41. Johansson E, Claesson R, van Dijken JW (2009) Antibacterial effect of ozone on cariogenic bacterial species. J Dent 37:449–453

    Article  PubMed  Google Scholar 

  42. Muller P, Guggenheim B, Schmidlin PR (2007) Efficacy of gasiform ozone and photodynamic therapy on a multispecies oral biofilm in vitro. Eur J Oral Sci 115:77–80

    Article  PubMed  Google Scholar 

  43. Doroszkiewicz W, Sikorska I, Jankowski S (1994) Studies on the influence of ozone on complement-mediated killing of bacteria. FEMS Immunol Med Microbiol 9:281–285

    Article  PubMed  Google Scholar 

  44. Schenkein HA, Genco RJ (1977) Gingival fluid and serum in periodontal diseases. II. Evidence for cleavage of complement components C3, C3 proactivator (factor B) and C4 in gingival fluid. J Periodontol 48:778–784

    PubMed  Google Scholar 

  45. Potempa M, Potempa J, Kantyka T, Nguyen KA, Wawrzonek K, Manandhar SP, Popadiak K, Riesbeck K, Eick S, Blom AM (2009) Interpain A, a cysteine proteinase from Prevotella intermedia, inhibits complement by degrading complement factor C3. PLoS Pathog 5:e1000316

    Article  PubMed  Google Scholar 

  46. Potempa M, Potempa J, Okroj M, Popadiak K, Eick S, Nguyen KA, Riesbeck K, Blom AM (2008) Binding of complement inhibitor C4b-binding protein contributes to serum resistance of Porphyromonas gingivalis. J Immunol 181:5537–5544

    PubMed  Google Scholar 

  47. Taubman MA, Valverde P, Han X, Kawai T (2005) Immune response: the key to bone resorption in periodontal disease. J Periodontol 76:2033–2041

    Article  PubMed  Google Scholar 

  48. Cochran DL (2008) Inflammation and bone loss in periodontal disease. J Periodontol 79:1569–1576

    Article  PubMed  Google Scholar 

  49. Huth KC, Saugel B, Jakob FM, Cappello C, Quirling M, Paschos E, Ern K, Hickel R, Brand K (2007) Effect of aqueous ozone on the NF-kappaB system. J Dent Res 86:451–456

    Article  PubMed  Google Scholar 

  50. Ducusin RJ, Nishimura M, Sarashina T, Uzuka Y, Tanabe S, Otani M (2003) Phagocytosis of bovine blood and milk polymorphonuclear leukocytes after ozone gas administration in vitro. J Vet Med Sci 65:535–539

    Article  PubMed  Google Scholar 

  51. Huth KC, Jakob FM, Saugel B, Cappello C, Paschos E, Hollweck R, Hickel R, Brand K (2006) Effect of ozone on oral cells compared with established antimicrobials. Eur J Oral Sci 114:435–440

    Article  PubMed  Google Scholar 

Download references

Acknowledgments

The authors are grateful to Regula Hirschi and Marianne Weibel for excellent assistance in performing the in vitro assays. This study was supported by W&H (W&H, Bürmoos, Austria).

Conflict of interest

The authors declare that they have no conflict of interest.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Sigrun Eick.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Eick, S., Tigan, M. & Sculean, A. Effect of ozone on periodontopathogenic species—an in vitro study. Clin Oral Invest 16, 537–544 (2012). https://doi.org/10.1007/s00784-011-0515-1

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00784-011-0515-1

Keywords

Navigation