İrrigasyon Rejimlerinin Enterococcus Faecalis Biofilmi Üzerine İn-Vitro Antibakteriyel Etkinliği

Year 2013, Volume: 23 Issue: 2, 165 - 171, 01.02.2013

Dr. Didem Atabek Yrd. Doç. Dr. Çağdaş Çınar Prof. Dr. Nurhan Öztaş Yrd. Doç. Dr. Gülçin Akca. Prof. Dr. Zek Suludere

Abstract

Objectives: The aim of this study was to compare the antibacterial efficiency of 2.25% sodium hypochlorite (NaOCl), 2% chlorhexidine (CHX), and gaseous ozone to biofilm of Enterococcus faecalis ( E. faecalis) generated on cellulose nitrate membrane filters. Materials and methods: After an overnight culture of E. faecalis grown in tryptic soy (TYC) broth, an aliquot of 100µl of microorganisms was seeded onto 13.0-mm diameter cellulose nitrate membrane filters (Sartorius®, UK) and incubated for 48 to 72 h at 37ºC in an aerobic atmosphere. Twenty membranes were used for each test agent. Then, biofilm formation test agents, gaseous ozone, and 0.9% NaCl (positive control) were applied at time intervals. Application periods for NaOCl and CHX were 5 to 10 minutes, whereas ozone application with Ozonytronx (Bizonix, GmbH, Germany) was 80 to 120 seconds. Next, from each of the samples, 100µl aliquots were put onto the TYC agar plates. Incubation colonies were calculated as colony forming units (CFU/mL) per membrane and observed by scanning electron microscopy (SEM). Data were analyzed using the Wilcoxon test, KruskalWallis test, and Mann-Whitney U-test at p Results: All of the irrigation regimens disgregated and removed the biofilm compared to the positive control (p CHX caused a significantly higher degree of biofilm disgregation than ozone application (p Conclusions: The irrigation regimen of 2.25% NaOCl and 2% CHX was more effective in eliminating biofilm formation of E. faecalis. However, the long-term irrigation regimen of ozone was found to be a successful alternative for endodontic treatment.Key-words: Biofilm, Ozone, Enterococcus faecalis

Keywords

-

In-Vitro Antibacterial Efficiency Of Irrigation Regimens Against Biofilm Of Enterococcus Faecalis

Abstract

Amaç: Çalışmanın amacı % 2,25 sodyum hipoklorid (NaOCl), %2 klorheksidin (CHX) ve ozon gazının selüloz nitrat membran filtreler üzerinde oluşturulan Enterococcus faecalis ( E. faecalis) biyofilmi üzerindeki antibakteriyal etkinliğini karşılaştırmaktır. Materyal method: Triptik soy (TYC) besiyerinde bir gece E. faecalis kültüre edilerek, 100 µl mikroorganizma sıvısı 13,0-mm çapında selüloz nitrat membran filtreler (Sartorius®, İngiltere) üzerine ekildi ve 48-72 saat 37ºC’de aerobic bir atmosferde inkübe edildi. Her test ajanı için 20 membran kullanıldı. Test ajanları, ozon gazı ve %0,9 NaCl (pozitif kontrol) zaman aralıklarında biyofilm formasyonuna uygulandı. NaOCl ve CHX için uygulama periyotları 5 ve 10 dak. iken; Ozonytronx (Bizonix, GmbH, Almanya) cihazı ile ozon gazı 80 ve 120 saniye uygulandı. Her örnekten 100µl sıvı TYC agarlara konuldu. İnkübasyon kolonileri membran yüzeyinde koloni oluşturan birimler (CFU/mL) olarak hesaplandı ve SEM ile gözlemlendi. Veriler Wilcoxon, Kruskal-Wallis ve Mann-Whitney Utestleri kullanılarak p<0,05 güven aralığında değerlendirildi. Bulgular: Pozitif kontrol grubu ile kıyaslandığında tüm irrigasyon rejimleri biyofilmi bozuntuya uğrattı ( p CHX ozon grubundan istatistiksel olarak anlamlı farklılık yaratacak düzeyde yüksek biyofilm bozunumu oluşturdu (p Sonuç: %2,25 NaOCl ve %2 CHX rejimleri E. faecalis biyofilm formasyonunun eliminasyonunda daha etkilidir. Öte yandan daha uzun süreli ozon gazı uygulamaları endodontik tedavi için alternatif bir irrigasyon rejimi olabilir.

Keywords

Biofilm, Ozon, Enterococcus faecalis

References

• Hems RS, Gulabivala K, Ng YL, Ready D, Spratt DA. An in vitro evaluation of the ability of ozone to kill a strain of Enterococcus faecalis. Int Endod J 2005; 38: 22-9.
• Byström A, Sundqvist G. Bacteriologic evaluation of the effect of 0.5 percent sodium hypochlorite in endodontic therapy. Oral Surg Oral Med Oral Pathol 1983; 55:307-12.
• Arias-Moliz MT, Ferrer-Luque CM, EspigaresGarcía M, Baca P. Enterococcus faecalis biofilms eradication by root canal irrigants. J Endod 2009; 35:711-4.
• Lin LM, Skribner JE, Gaengler P. Factors associated with endodontic treatment failures. J Endod 1992; 18:625-7.
• Giardino L, Ambu E, Savoldi E, Rimondini R, Cassanelli C, Debbia EA. Comparative evaluation of antimicrobial efficacy of sodium hypochlorite, MTAD, and Tetraclean against Enterococcus faecalis biofilm. J Endod 2007; 33:852-5.
• Hope CK, Garton SG, Wang Q, Burnside G, Farrelly PJ. A direct comparison between extracted tooth and filter-membrane biofilm models of endodontic irrigation using Enterococcus faecalis. Arch Microbiol 2010; 192: 775- Epub 2010 Jul 23.
• Huth KC, Quirling M, Maier S, Kamereck K, Alkhayer M, Paschos E, Welsch U, Miethke T, Brand K, Hickel R. Effectiveness of ozone against endodontopathogenic microorganisms in a root canal biofilm model. Int Endod J 2009; 42:3-13.
• Grigoratos D, Knowles J, Ng YL, Gulabivala K. Effect of exposing dentine to sodium hypochlorite and calcium hydroxide on its flexural strength and elastic modulus. Int Endod J 2001; 34:113-9.
• Sim TP, Knowles JC, Ng YL, Shelton J, Gulabivala K. Effect of sodium hypochlorite on mechanical properties of dentine and tooth surface strain. Int Endod J 2001; 34:120-32.
• Gernhardt CR, Eppendorf K, Kozlowski A, Brandt M. Toxicity of concentrated sodium hypochlorite used as an endodontic irrigant. Int Endod J 2004; 37:272-80.
• Estrela C, Silva JA, de Alencar AH, Leles CR, Decurcio DA. Efficacy of sodium hypochlorite and chlorhexidine against Enterococcus faecalis--a systematic review. J Appl Oral Sci 2008;16:364
• Hofer D, Meier A, Sener B, Guggenheim B, Attin T, Schmidlin PR. Biofilm reduction and staining potential of a 0.05% chlorhexidine rinse containing essential oils. Int J Dent Hyg 2011; 9:60Azarpazhooh A, Limeback H. The application of ozone in dentistry: a systematic review of literature. J Dent 2008; 36:104-16. Epub 2007 Dec
• Knight GM, McIntyre JM, Craig GG, Mulyani. Ion uptake into demineralized dentine from glass ionomer cement following pretreatment with silver fluoride and potassium iodide. Aust Dent J 2006; 51:237-41.
• Dunavant TR, Regan JD, Glickman GN, Solomon ES, Honeyman AL. Comparative evaluation of endodontic irrigants against Enterococcus faecalis biofilms. J Endod 2006; 32:527-31.
• Gomes BP, Ferraz CC, Vianna ME, Berber VB, Teixeira FB, Souza-Filho FJ. In vitro antimicrobial activity of several concentrations of sodium hypochlorite and chlorhexidine gluconate in the elimination of Enterococcus faecalis. Int Endod J 2001; 34:424-8.
• Zerella JA, Fouad AF, Spångberg LS. Effectiveness of a calcium hydroxide and chlorhexidine digluconate mixture as disinfectant during retreatment of failed endodontic cases. Oral Surg Oral Med Oral Pathol Oral Radiol Endod 2005; 100:756-61.
• Huth KC, Saugel B, Jakob FM, Cappello C, Quirling M, Paschos E, Ern K, Hickel R, Brand K. Effect of aqueous ozone on the NF-kappaB system. J Dent Res 2007; 86:451-6.
• Molander A, Reit C, Dahlén G, Kvist T. Microbiological status of root-filled teeth with apical periodontitis. Int Endod J 1998; 31:1-7.
• Baca P, Junco P, Arias-Moliz MT, GonzálezRodríguez MP, Ferrer-Luque CM. Residual and antimicrobial activity of final irrigation protocols on Enterococcus faecalis biofilm in dentin. J Endod 2011; 37:363-6.
• Nagayoshi M, Kitamura C, Fukuizumi T, Nishihara T, Terashita M. Antimicrobial effect of ozonated water on bacteria invading dentinal tubules. J Endod 2004; 30:778-81.
• Cardoso MG, de Oliveira LD, Koga-Ito CY, Jorge AO. Effectiveness of ozonated water on Candida albicans, Enterococcus faecalis, and endotoxins in root canals. Oral Surg Oral Med Oral Pathol Oral Radiol Endod 2008; 105:e85-91.
• Estrela C, Estrela CR, Decurcio DA, Hollanda AC, Silva JA. Antimicrobial efficacy of ozonated water, gaseous ozone, sodium hypochlorite and chlorhexidine in infected human root canals. Int Endod J 2007; 40:85-93.
• Onçağ O, Hoşgör M, Hilmioğlu S, Zekioğlu O, Eronat C, Burhanoğlu D. Comparison of antibacterial and toxic effects of various root canal irrigants. Int Endod J 2003; 36:423-32.
• Hubbezoğlu İ, Ülkü Ö, Sümer Z. Kök kanal irrigasyonunda propolis içerikli solüsyonlar ile potasyum titanyum fosfat lazerin Esherichia Coli üzerine etkileri. Atatürk Üniv Diş Hek Fak Derg 2011; 21:15-21 Yazışma Adresi: Didem ATABEK, PhD
• Gazi University, Faculty of Dentistry, Department of Pedodontics, Ankara/TURKEY,06510 Tel: [phone] Fax: [phone]
• E-mail: [e-mail]