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Clinical antibacterial effectiveness and biocompatibility of gaseous ozone after incomplete caries removal

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Abstract

Objectives

To evaluate local effect of gaseous ozone on bacteria in deep carious lesions after incomplete caries removal, using chlorhexidine as control, and to investigate its effect on pulp vascular endothelial growth factor (VEGF), neuronal nitric oxide synthase (nNOS), and superoxide dismutase (SOD).

Materials and methods

Antibacterial effect was evaluated in 48 teeth with diagnosed deep carious lesion. After incomplete caries removal, teeth were randomly allocated into two groups regarding the cavity disinfectant used: ozone (open system) or 2% chlorhexidine. Dentin samples were analyzed for the presence of total bacteria and Lactobacillus spp. by real-time quantitative polymerase chain reaction. For evaluation of ozone effect on dental pulp, 38 intact permanent teeth indicated for pulp removal/tooth extraction were included. After cavity preparation, teeth were randomly allocated into two groups: ozone group and control group. VEGF/nNOS level and SOD activity in dental pulp were determined by enzyme-linked immunosorbent assay and spectrophotometric method, respectively.

Results

Ozone application decreased number of total bacteria (p = 0.001) and Lactobacillus spp. (p < 0.001), similarly to chlorhexidine. The VEGF (p < 0.001) and nNOS (p = 0.012) levels in dental pulp after ozone application were higher, while SOD activity was lower (p = 0.001) comparing to those in control pulp.

Conclusions

Antibacterial effect of ozone on residual bacteria after incomplete caries removal was similar to that of 2% chlorhexidine. Effect of ozone on pulp VEGF, nNOS, and SOD indicated its biocompatibility.

Clinical relevance

Ozone appears as effective and biocompatible cavity disinfectant in treatment of deep carious lesions by incomplete caries removal technique.

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References

  1. Schwendicke F, Dorfer CE, Paris S (2013) Incomplete caries removal: a systematic review and meta-analysis. J Dent Res 92:306–314. https://doi.org/10.1177/0022034513477425

    Article  PubMed  Google Scholar 

  2. Casagrande L, Seminario AT, Correa MB, Werle SB, Maltz M, Demarco FF, Araujo FB (2017) Longevity and associated risk factors in adhesive restorations of young permanent teeth after complete and selective caries removal: a retrospective study. Clin Oral Investig 21:847–855. https://doi.org/10.1007/s00784-016-1832-1

    Article  PubMed  Google Scholar 

  3. Gu F, Bresciani E, Barata TJ, Fagundes TC, Navarro MF, Dickens SH, Fenno JC, Peters MC (2010) In vivo acid etching effects on bacteria within caries-affected dentin. Caries Res 44:472–477. https://doi.org/10.1159/000320362

    Article  PubMed  PubMed Central  Google Scholar 

  4. Geerts S, Bolette A, Seidel L, Guéders A (2012) An in vitro evaluation of leakage of two etch and rinse and two self-etch adhesives after thermocycling. Int J Dent 2012:852841. https://doi.org/10.1155/2012/852841

    Article  PubMed  PubMed Central  Google Scholar 

  5. Hauser-Gerspach I, Pfäffli-Savtchenko V, Dähnhardt JE, Meyer J, Lussi A (2009) Comparison of the immediate effects of gaseous ozone and chlorhexidine gel on bacteria in cavitated carious lesions in children in vivo. Clin Oral Investig 13:287–291. https://doi.org/10.1007/s00784-008-0234-4

    Article  PubMed  Google Scholar 

  6. Bin-Shuwaish MS (2016) Effects and effectiveness of cavity disinfectants in operative dentistry: a literature review. J Contemp Dent Pract 17:867–879

    Article  Google Scholar 

  7. Dukić W, Jurić H, Andrašević AT, Kovačević V, Dukić OL, Delija B (2013) The efficacy of gaseous ozone on some cariogenic bacteria. Coll Antropol 37:109–113

    PubMed  Google Scholar 

  8. Castillo A, Galindo-Moreno P, Avila G, Valderrama M, Liébana J, Baca P (2008) In vitro reduction of mutans streptococci by means of ozone gas application. Quintessence Int 39:827–831

    PubMed  Google Scholar 

  9. Johansson E, Claesson R, van Dijken JW (2009) Antibacterial effect of ozone on cariogenic bacterial species. J Dent 37:449–453. https://doi.org/10.1016/j.jdent.2009.02.004

    Article  PubMed  Google Scholar 

  10. Polydorou O, Halili A, Wittmer A, Pelz K, Hahn P (2012) The antibacterial effect of gas ozone after 2 months of in vitro evaluation. Clin Oral Investig 16:545–550. https://doi.org/10.1007/s00784-011-0524-0

    Article  PubMed  Google Scholar 

  11. Kollmuss M, Kist S, Obermeier K, Pelka AK, Hickel R, Huth KC (2014) Antimicrobial effect of gaseous and aqueous ozone on caries pathogen microorganisms grown in biofilms. Am J Dent 27:134–138

    PubMed  Google Scholar 

  12. Cadenaro M, Delise C, Antoniollo F, Navarra OC, Di Lenarda R, Breschi L (2009) Enamel and dentin bond strength following gaseous ozone application. J Adhes Dent 11:287–292

    PubMed  Google Scholar 

  13. Borges GÁ, Elias ST, da Silva SM, Magalhães PO, Macedo SB, Ribeiro AP, Guerra EN (2017) In vitro evaluation of wound healing and antimicrobial potential of ozone therapy. J Craniomaxillofac Surg 45:364–370. https://doi.org/10.1016/j.jcms.2017.01.005.

    Article  PubMed  Google Scholar 

  14. Taşdemir Z, Alkan BA, Albayrak H (2016) Effects of ozone therapy on the early healing period of deepithelialized gingival grafts: a randomized placebo-controlled clinical trial. J Periodontol 87:663–671. https://doi.org/10.1902/jop.2016.150217

    Article  PubMed  Google Scholar 

  15. Akdeniz SS, Beyler E, Korkmaz Y, Yurtcu E, Ates U, Araz K, Sahin FI, Torun OY (2017) The effects of ozone application on genotoxic damage and wound healing in bisphosphonate-applied human gingival fibroblast cells. Clin Oral Investig 22:867–873. https://doi.org/10.1007/s00784-017-2163-6

    Article  PubMed  Google Scholar 

  16. Kim HS, Noh SU, Han YW, Kim KM, Kang H, Kim HO, Park JM (2009) Therapeutic effects of topical application of ozone on acute cutaneous wound healing. J Korean Med Sci 24:368–374. https://doi.org/10.3346/jkms.2009.24.3.368

    Article  PubMed  PubMed Central  Google Scholar 

  17. Zhang J, Guan M, Xie C, Luo X, Zhang Q, Xue Y (2014) Increased growth factors play a role in wound healing promoted by noninvasive oxygen-ozone therapy in diabetic patients with foot ulcers. Oxidative Med Cell Longev 2014:273475. https://doi.org/10.1155/2014/273475

    Article  Google Scholar 

  18. Valacchi G, Bocci V (2000) Studies on the biological effects of ozone: 11. Release of factors from human endothelial cells. Mediat Inflamm 9:271–276

    Article  Google Scholar 

  19. Yıldırım AO, Eryılmaz M, Kaldırım U, Eyi YE, Tuncer SK, Eroğlu M, Durusu M, Topal T, Kurt B, Dilmen S, Bilgiç S, Serdar M (2014) Effectiveness of hyperbaric oxygen and ozone applications in tissue healing in generated soft tissue trauma model in rats: an experimental study. Ulus Travma Acil Cerrahi Derg 20:167–175. https://doi.org/10.5505/tjtes.2014.09465

    Article  PubMed  Google Scholar 

  20. Ferrara N, Gerber HP, LeCouter J (2003) The biology of VEGF and its receptors. Nat Med 9:669–676. https://doi.org/10.1038/nm0603-669

    Article  PubMed  Google Scholar 

  21. Förstermann U, Sessa WC (2012) Nitric oxide synthases: regulation and function. Eur Heart J 33:829–837. https://doi.org/10.1093/eurheartj/ehr304

    Article  PubMed  Google Scholar 

  22. Nadkarni MA, Martin FE, Jacques NA, Hunter N (2002) Determination of bacterial load by real time PCR using a broad-range (universal) probe and primers set. Microbiology 148:257–266. https://doi.org/10.1099/00221287-148-1-257

    Article  PubMed  Google Scholar 

  23. Byun R, Nadkarni MA, Chhour KL, Martin FE, Jacques NA, Hunter N (2004) Quantitative analysis of diverse lactobacillus species present in advanced dental caries. J Clin Microbiol 42:3128–3136. https://doi.org/10.1128/JCM.42.7.3128-3136.2004

    Article  PubMed  PubMed Central  Google Scholar 

  24. Lafuente D (2012) SEM analysis of hybrid layer and bonding interface after chlorhexidine use. Oper Dent 37:172–180. https://doi.org/10.2341/10-251-L

    Article  PubMed  Google Scholar 

  25. Matsuzaka K, Muramatsu T, Katakura A, Ishihara K, Hashimoto S, Yoshinari M, Endo T, Tazaki M, Shintani M, Sato Y, Inoue T (2008) Changes in the homeostatic mechanism of dental pulp with age: expression of the core-binding factor alpha-1, dentin sialoprotein, vascular endothelial growth factor, and heat shock protein 27 messenger RNAs. J Endod 34:818–821. https://doi.org/10.1016/j.joen.2008.03.027

    Article  PubMed  Google Scholar 

  26. Davis WL, Jacoby BH, Craig KR, Wagner G, Harrison JW (1991) Copper-zinc superoxide dismutase activity in normal and inflamed human dental pulp tissue. J Endod 17:316–318. https://doi.org/10.1016/S0099-2399(06)81698-5

    Article  PubMed  Google Scholar 

  27. Yu C, Abbott PV (2007) An overview of the dental pulp: its functions and responses to injury. Aust Dent J 52(1 Suppl):S4–S16

    Article  Google Scholar 

  28. Ilić J, Radović K, Roganović J, Brković B, Stojić D (2012) The levels of vascular endothelial growth factor and bone morphogenetic protein 2 in dental pulp tissue of healthy and diabetic patients. J Endod 38:764–768. https://doi.org/10.1016/j.joen.2012.03.016

    Article  PubMed  Google Scholar 

  29. Virtej A, Løes S, Iden O, Bletsa A, Berggreen E (2013) Vascular endothelial growth factors signalling in normal human dental pulp: a study of gene and protein expression. Eur J Oral Sci 121:92–100. https://doi.org/10.1111/eos.12019

    Article  PubMed  Google Scholar 

  30. Artese L, Corrado R, Ferrero G, Fiorini M, Santinelli A, Piatelli A (2002) Vascular endothelial growth factor (VEGF) expression in healthy and inflamed human dental pulps. J Endod 28:20–23. https://doi.org/10.1097/00004770-200201000-00005

    Article  PubMed  Google Scholar 

  31. Tran-Hung L, Laurent P, Camps J, About I (2008) Quantification of angiogenic growth factors released by human dental cells after injury. Arch Oral Biol 53:9–13

    Article  Google Scholar 

  32. Scheven BA, Man J, Millard JL, Cooper PR, Lea SC, Walmsley AD, Smith AJ (2008) VEGF and odontoblast-like cells: stimulation by low frequency ultrasound. Arch Oral Biol 54:185–191. https://doi.org/10.1016/j.archoralbio.2008.09.008

    Article  PubMed  Google Scholar 

  33. Ishida S, Usui T, Yamashiro K, Kaji Y, Amano S, Ogura Y, Hida T, Oguchi Y, Ambati J, Miller JW, Gragoudas ES, Ng YS, D'Amore PA, Shima DT, Adamis AP (2003) VEGF164-mediated inflammation is required for pathological, but not physiological, ischemia-induced retinal neovascularization. J Exp Med 198:483–489. https://doi.org/10.1084/jem.20022027

    Article  PubMed  PubMed Central  Google Scholar 

  34. Mantellini MG, Botero T, Yaman P, Dennison JB, Hanks CT, Nör JE (2006) Adhesive resin and the hydrophilic monomer HEMA induce VEGF expression on dental pulp cells and macrophages. Dent Mater 22:434–440. https://doi.org/10.1016/j.dental.2005.04.039

    Article  PubMed  Google Scholar 

  35. Mattuella LG, de Figueiredo JA, Nör JE, de Araujo FB, Fossati AC (2007) Vascular endothelial growth factor receptor–2 expression in the pulp of human primary and young permanent teeth. J Endod 33:1408–1412. https://doi.org/10.1016/j.joen.2007.08.019

    Article  Google Scholar 

  36. Korkmaz Y, Baumann MA, Steinritz D, Schröder H, Behrends S, Addicks K, Schneider K, Raab WH, Bloch W (2005) NO-cGMP signaling molecules in cells of the rat molar dentin-pulp complex. J Dent Res 84:618–623. https://doi.org/10.1177/154405910508400707

    Article  PubMed  Google Scholar 

  37. Cupertino RR, Fabri FV, Veltrini VC, Hidalgo MM, Bruschi ML, de Oliveira RM (2016) Histological evaluation of the rat dental pulp after indirect capping with sildenafil or L-NAME incorporated into a bioadhesive thermoresponsive system. Acta Sci Health Sci 38:95

    Article  Google Scholar 

  38. Lohinai Z, Székely AD, Benedek P, Csillag A (1997) Nitric oxide synthase containing nerves in the cat and dog dental pulp and gingiva. Neurosci Lett 227:91–94

    Article  Google Scholar 

  39. Leone A, Patel M, Uzzo ML, Buscemi M, Gerbino A (2002) Expression and modification of NO synthase in human dental pulps during orthodontic treatment. Bull Group Int Rech Sci Stomatol Odontol 44:57–60

    PubMed  Google Scholar 

  40. Di Nardo Di Maio F, Lohinai Z, D’Arcangelo C, De Fazio PE, Speranza L, De Lutiis MA, Patruno A, Grilli A, Felaco M (2004) Nitric oxide synthase in healthy and inflamed human dental pulp. J Dent Res 83:312–316. https://doi.org/10.1177/154405910408300408

    Article  PubMed  Google Scholar 

  41. Silva Mendez LS, Allaker RP, Hardie JM, Benjamin N (1999) Antimicrobial effect of acidified nitrite on cariogenic bacteria. Oral Microbiol Immunol 14:391–392

    Article  Google Scholar 

  42. Farges JC, Bellanger A, Ducret M, Aubert-Foucher E, Richard B, Alliot-Licht B, Bleicher F, Carrouel F (2015) Human odontoblast-like cells produce nitric oxide with antibacterial activity upon TLR2 activation. Front Physiol 6:185. https://doi.org/10.3389/fphys.2015.00185

    Article  PubMed  PubMed Central  Google Scholar 

  43. Al-Omiri MK, Alhijawi M, AlZarea BK, Abul Hassan RS, Lynch E (2016) Ozone treatment of recurrent aphthous stomatitis: a double blinded study. Sci Rep 6:27772. https://doi.org/10.1038/srep27772

    Article  PubMed  PubMed Central  Google Scholar 

  44. Sivalingam VP, Panneerselvam E, Raja KV, Gopi G (2017) Does topical ozone therapy improve patient comfort after surgical removal of impacted mandibular third molar? A randomized controlled trial. J Oral Maxillofac Surg 75:51.e1–51.e9. https://doi.org/10.1016/j.joms.2016.09.014

    Article  Google Scholar 

  45. Kawabata A, Manabe S, Manabe Y, Takagi H (1994) Effect of topical administration of L-arginine on formalin-induced nociception in the mouse: a dual role of peripherally formed NO in pain modulation. Br J Pharmacol 112:547–550

    Article  Google Scholar 

  46. Sagai M, Bocci V (2011) Mechanisms of action involved in ozone therapy: is healing induced via a mild oxidative stress? Med Gas Res 1:29. https://doi.org/10.1186/2045-9912-1-29

    Article  PubMed  PubMed Central  Google Scholar 

  47. Greene AK, Güzel-Seydim ZB, Seydim AC (2012) Chemical and physical properties of ozone. In: O’Donnell C, Tiwari BK, Cullen PJ, Rice RG (eds) Ozone in food processing. Wiley-Blackwell, Oxford, p 26

    Google Scholar 

  48. Varvara G, Traini T, Esposito P, Caputi S, Perinetti G (2005) Copper-zinc superoxide dismutase activity in healthy and inflamed human dental pulp. Int Endod J 38:195–199. https://doi.org/10.1111/j.1365-2591.2005.00936.x

    Article  PubMed  Google Scholar 

  49. Bödör C, Matolcsy A, Bernáth M (2007) Elevated expression of Cu, Zn-SOD and Mn-SOD mRNA in inflamed dental pulp tissue. Int Endod J 40:128–132. https://doi.org/10.1111/j.1365-2591.2006.01196.x

    Article  PubMed  Google Scholar 

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Funding

This study was supported by the grant of Ministry of Science and Technology (No. 19/06-020/961-174/12), Republic of Srpska, Bosnia and Herzegovina.

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Correspondence to Jelena Krunić.

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All procedures performed in study were in accordance with the ethical standards of the Institutional research committee and with the 1964 Helsinki declaration and its later amendments.

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

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Krunić, J., Stojanović, N., Đukić, L. et al. Clinical antibacterial effectiveness and biocompatibility of gaseous ozone after incomplete caries removal. Clin Oral Invest 23, 785–792 (2019). https://doi.org/10.1007/s00784-018-2495-x

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  • DOI: https://doi.org/10.1007/s00784-018-2495-x

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