Skip to main content

Advertisement

SpringerLink
Log in

The effects of ozone application on genotoxic damage and wound healing in bisphosphonate-applied human gingival fibroblast cells

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

Abstract

Objectives

Medication-related osteonecrosis of the jaws (MRONJ) is an extremely therapy-resistant disease involving the jaws especially following bisphosphonate treatment. Bisphosphonates accumulate in bone in concentrations sufficient to be directly toxic to the oral epithelium. Current therapeutic options are inadequate for the prevention and treatment of MRONJ. The aim of this study was to investigate effects of ozone gas plasma therapy on wound healing in bisphosphonate-applied human fibroblasts.

Material and methods

Human primary gingival fibroblasts were cultured. Cytotoxic concentrations (IC50) of bisphosphonates (pamidronate (PAM), alendronate (ALN), and zoledronate (ZOL)) were determined by MTT test. A 60 μg/μl for 30 s of ozone gas plasma application was performed to all experimental culture flasks after drug treatment at 24-h intervals as 3 s/cm2. Genotoxic damages were evaluated by comet assay and wound healing was determined by in vitro scratch assay.

Results

PAM, ALN, and ZOL applications caused genotoxic damage on primary human gingival fibroblast DNA. Ozone gas plasma therapy significantly decreased the genotoxic damage (p < 0.05), and this application provided 25, 29, and 27% less genotoxic damage in order of ALN, PAM, and ZOL groups. Ozone gas plasma therapy significantly increased wound healing rates both in postsurgical 24th and 48th hours for all doses of experimental drug groups (p < 0.05).

Conclusion

The ozone gas plasma application decreased genotoxic damage effect of bisphosphonate usage while improved the wound closure rate on human gingival fibroblasts.

Clinical relevance

Ozone gas plasma therapy may be helpful in prevention of gingival healing delay in MRONJ pathogenesis especially when applied simultaneously with surgical intervention.

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

Access this article

Log in via an institution

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

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  1. Svejda B, Muschitz C, Gruber R, Brandtner C, Svejda C, Gasser RW, Santler G, Dimai HP (2016) Position paper on medication-related osteonecrosis of the jaw (MRONJ). Wien Med Wochenschr 166:68–74

    Article  PubMed  Google Scholar 

  2. Soydan SS, Araz K, Senel FV, Yurtcu E, Helvacioglu F, Dagdeviren A, Tekindal MA, Sahin F (2015) Effects of alendronate and pamidronate on apoptosis and cell proliferation in cultured primary human gingival fibroblasts. Hum Exp Toxicol 34:1073–1082

    Article  PubMed  Google Scholar 

  3. Ravosa MJ, Ning J, Liu Y, Stack MS (2011) Bisphosphonate effects on the behaviour of oral epithelial cells and oral fibroblasts. Arch Oral Biol 56:491–498

    Article  PubMed  Google Scholar 

  4. Walter C, Klein MO, Pabst A, Al-Nawas B, Duschner H, Ziebart T (2009) Influence of bisphosphonates on endothelial cells, fibroblasts, and osteogenic cells. Clin Oral Investig 14:35–41

    Article  PubMed  Google Scholar 

  5. Pabst AM, Ziebart T, Koch FP, Taylor KY, Al-Nawas B, Walter C (2012) The influence of bisphosphonates on viability, migration, and apoptosis of human oral keratinocytes—in vitro study. Clin Oral Investig 16:87–93

    Article  PubMed  Google Scholar 

  6. Simon MJ, Niehoff P, Kimmig B, Wiltfang J, Açil Y (2010) Expression profile and synthesis of different collagen types I, II, III, and V of human gingival fibroblasts, osteoblasts, and SaOS-2 cells after bisphosphonate treatment. Clin Oral Investig 14:51–58

    Article  PubMed  Google Scholar 

  7. Acil Y, Moller B, Niehoff P, Rachko K, Gassling V, Wiltfang J, Simon MJ (2012) The cytotoxic effects of three different bisphosphonates in-vitro on human gingival fibroblasts, osteoblasts and osteogenic sarcoma cells. J Craniomaxillofac Surg 40:e229–e235

    Article  PubMed  Google Scholar 

  8. Cozin M, Pinker BM, Solemani K, Zuniga JM, Dadaian SC, Cremers S, Landesberg R, Raghavan S (2011) Novel therapy to reverse the cellular effects of bisphosphonates on primary human oral fibroblasts. J Oral Maxillofac Surg 69:2564–2578

    Article  PubMed  PubMed Central  Google Scholar 

  9. Nogales CG, Ferrari PH, Kantorovich EO, Lage-Marques J (2008) Ozone therapy in medicine and dentistry. J Contemp Dental Pract 9:75–84

    Google Scholar 

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

    Article  Google Scholar 

  11. Zimmermann D, Waltimo T, Filippi A (2012) Ozonized water in dental traumatology—a preliminary study on the treatment of avulsed teeth, in vitro. Ozone Sci Eng 34:484–488

    Article  Google Scholar 

  12. Yurtcu E, İşeri Ö, Sahin F (2014) Genotoxic and cytotoxic effects of doxorubicin and silymarin on human hepatocellular carcinoma cells. Hum Exp Toxicol 33:1269–1276

    Article  PubMed  Google Scholar 

  13. Işeri OD, Sahin FI, Terzi YK, Yurtcu E, Erdem SR, Sarialioglu F (2014) beta-Adrenoreceptor antagonists reduce cancer cell proliferation, invasion, and migration. Pharm Biol 52:1374–1381

    Article  PubMed  Google Scholar 

  14. Bocci V (2004) Ozone as Janus: this controversial gas can be either toxic or medically useful. Mediat Inflamm 13:3–11

    Article  Google Scholar 

  15. Domb WC (2014) Ozone therapy in dentistry: a brief review for physicians. Interv Neuroradiol 20:632–636

    Article  PubMed  PubMed Central  Google Scholar 

  16. Erdemci F, Gunaydin Y, Sencimen M, Bassorgun I, Ozler M, Oter S, Gulses A, Gunal A, Sezgin S, Bayar GR, Dogan N, Gider IK (2014) Histomorphometric evaluation of the effect of systemic and topical ozone on alveolar bone healing following tooth extraction in rats. Int J Oral Maxillofac Surg 43:777–783

    Article  PubMed  Google Scholar 

  17. Steinhart H, Schulz S, Mutters R (1999) Evaluation of ozonated oxygen in an experimental animal model of osteomyelitis as a further treatment option for skull-base osteomyelitis. Eur Arch Otorhinolaryngol 256:153–157

    Article  PubMed  Google Scholar 

  18. Agrillo A, Filiaci F, Ramieri V, Riccardi E, Quarato D, Rinna C, Gennaro P, Cascino F, Mitro V, Ungari C (2012) Bisphosphonate-related osteonecrosis of the jaw (BRONJ): 5 year experience in the treatment of 131 cases with ozone therapy. Eur Rev Med Pharmacol Sci 16:1741–1747

    PubMed  Google Scholar 

  19. Agrillo A, Ungari C, Filiaci F, Priore P, Iannetti G (2007) Ozone therapy in the treatment of avascular bisphosphonate-related jaw osteonecrosis. J Craniofac Surg 18:1071–1075

    Article  PubMed  Google Scholar 

  20. Petrucci MT, Gallucci C, Agrillo A, Mustazza MC, Foà R (2007) Role of ozone therapy in the treatment of osteonecrosis of the jaws in multiple myeloma patients. Haematologica 92:1289–1290

    Article  PubMed  Google Scholar 

  21. Vescovi P, Merigo E, Meleti M, Manfredi M, Fornaini C, Nammour S, Mergoni G, Sarraj A, Bagan JV (2014) Conservative surgical management of stage I bisphosphonate-related osteonecrosis of the jaw. Int J Dent 2014:107690. doi:10.1155/2014/107690

    Article  PubMed  PubMed Central  Google Scholar 

  22. Ripamonti CI, Cislaghi E, Mariani L, Maniezzo M (2011) Efficacy and safety of medical ozone (O(3)) delivered in oil suspension applications for the treatment of osteonecrosis of the jaw in patients with bone metastases treated with bisphosphonates: preliminary results of a phase I–II study. Oral Oncol 47:185–190

    Article  PubMed  Google Scholar 

  23. Agrillo A, Sassano P, Rinna C, Priore P, Iannetti G (2007) Ozone therapy in extractive surgery on patients treated with bisphosphonates. J Craniofac Surg 18:1068–1070

    Article  PubMed  Google Scholar 

  24. Agrillo A, Petrucci MT, Tedaldi M, Mustazza MC, Marino SMF, Gallucci C, Iannetti G (2006) New therapeutic protocol in the treatment of avascular necrosis of the jaws. J Craniofac Surg 17:1080–1083

    Article  PubMed  Google Scholar 

  25. Csonka C, Pataki T, Kovacs P, Müller SL, Schroeter ML, Tosaki A, Blasig IE (2000) Effects of oxidative stress on the expression of antioxidative defense enzymes in spontaneously hypertensive rat hearts. Free Radic Biol Med 29:612–619

    Article  PubMed  Google Scholar 

  26. Clavo B, Catalá L, Pérez JL, Rodríguez V, Robaina F (2004) Ozone therapy on cerebral blood flow: a preliminary report. Evid Based Complement Alternat Med 1:315–319

    Article  PubMed  PubMed Central  Google Scholar 

  27. Bocci V (1999) Biological and clinical effects of ozone. Has ozone therapy a future in medicine? Br J Biomed Sci 56:270–279

    PubMed  Google Scholar 

  28. Menendez S, Falcon L, Simon DR, Landa N (2002) Efficacy of ozonized sunflower oil in the treatment of tinea pedis. Mycoses 45:329–332

    Article  PubMed  Google Scholar 

  29. Baysan A, Whiley RA, Lynch E (2000) Antimicrobial effect of a novel ozone generating device on micro-organisms associated with primary root carious lesions in vitro. Caries Res 34:498–501

    Article  PubMed  Google Scholar 

  30. Vannucchi AM, Ficarra G, Antonioli E, Bosi A (2005) Osteonecrosis of the jaw associated with zoledronate therapy in a patient with multiple myeloma. Br J Haematol 128:738

    Article  PubMed  Google Scholar 

  31. Alpan AL, Toker H, Ozer H (2016) Ozone therapy enhances osseous healing in diabetic rats with calvarial defect: a morphometric and immunohistochemical study. J Periodontol 87:982–989

    Article  PubMed  Google Scholar 

  32. Taşdemir Z, Alkan BA, Albayrak H (2016) The effects of ozone therapy on the early healing period of de-epithelialized gingival grafts: a randomized placebo-controlled clinical trial. J Periodontol 87:663–671

    Article  PubMed  Google Scholar 

  33. Al Habashneh R, Alsalman W, Khader Y (2015) Ozone as an adjunct to conventional nonsurgical therapy in chronic periodontitis: a randomized controlled clinical trial. J Periodontal Res 50:37–43

    Article  PubMed  Google Scholar 

  34. Marx RE, Sawatary Y, Fortin M, Broumand V (2005) Bisphosphonate induced exposed bone (osteonecrosis/osteopetrosis) of the jaws: risk factors, recognition, prevention, and treatment. J Oral Maxillofac Surg 63:1567–1575

    Article  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Oral and Maxillofacial Surgery, Faculty of Dentistry, Baskent University, Bahçelievler 11. Sokak no:26 Mareşal Fevzi Çakmak Caddesi Çankaya, 06490, Ankara, Turkey

    Sıdıka Sinem Akdeniz, E. Beyler & K. Araz

  2. Department of Medical Biology, Faculty of Medicine, Baskent University, Ankara, Turkey

    Y. Korkmaz & E. Yurtcu

  3. Private Practice, Ankara, Turkey

    U. Ates & O. Y. Torun

  4. Department of Medical Genetics, Faculty of Medicine, Baskent University, Ankara, Turkey

    F. I. Sahin

Authors
  1. Sıdıka Sinem Akdeniz
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. E. Beyler
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Y. Korkmaz
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. E. Yurtcu
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. U. Ates
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. K. Araz
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. F. I. Sahin
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. O. Y. Torun
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Sıdıka Sinem Akdeniz.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Funding

The work was financially supported by the Baskent University Institutional Review Board and Ethics Committee (Project No: D-DA 13/02).

Ethical approval

All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards.

Informed consent

Informed consent was obtained from all individual participants included in the study.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Akdeniz, S.S., Beyler, E., Korkmaz, Y. et al. The effects of ozone application on genotoxic damage and wound healing in bisphosphonate-applied human gingival fibroblast cells. Clin Oral Invest 22, 867–873 (2018). https://doi.org/10.1007/s00784-017-2163-6

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00784-017-2163-6

Keywords

Search

Navigation