Research Article
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IN VITRO COMPARISON OF THE ANTIMICROBIAL EFFECTS OF DIFFERENT ROOT CANAL MEDICAMENTS ON ENTEROCOCCUS FAECALIS AND CANDIDA ALBICANS

Year 2021, Volume: 24 Issue: 3, 256 - 265, 15.09.2021

Burçin Arıcan , Hesna Sazak Öveçoğlu

https://doi.org/10.7126/cumudj.912285

Abstract

Objectives: To evaluate the antimicrobial effects of experimental root canal medicaments on Enterococcus faecalis and Candida albicans on day 3 and day 7 using quantitative polymerase chain reaction (Q-PCR) analyses.
Materials and Methods: 240 single-rooted, single-canalled human teeth were used. Root canals were shaped mechanically and embedded in acrylic blocks, then sterilized in an autoclave. The samples were divided into two groups and infected with E. faecalis and C. albicans, and then divided into subgroups (n=10); calcium hydroxide (Ca(OH)2), triple paste (TAP), double paste (DP), modified double paste (MDP), lactoferrin (Lf), negative and positive groups. At the end of 3 and 7 days, paper points containing the root canal samples were placed in empty Eppendorf tubes, and DNA was isolated. Real-time Q-PCR was applied and the data were analyzed statistically.
Results: The antimicrobial effects of each medicine increased from days 3 to 7. Ca(OH)2 and TAP groups showed the similar eradication rates for E. faecalis and C. albicans on day 3 and 7 (P > 0.05). There was no significant difference between DP, MDP and Lf for the eradication of microorganisms at both experimental days (P > 0.05), except the amount of eradicated E. faecalis by DP at day 7 in which DP caught the similar percentages with TAP and Ca(OH)2 (P < 0.05).
Conclusions: Experimental pastes demonstrated antimicrobial efficiancy similar to those used routinely in endodontic clinic. It is promising that lactoferrin which is a very biocompatible material can be used in different combinations as an intracanal medicament.

Keywords

Antibiotic paste Candida albicans Enterococcus faecalis Lactoferrin Root canal medicine

Supporting Institution

Marmara University Scientific Research Projects Committee

Project Number

SAG-C-DRP 150513–0154

Thanks

The authors would like to grateful for the financial support from Marmara University. The authors also thank Prof. Dr. Tanju Kadir and Dr. Banu Can.

References

  • 1. Gomes BPFA, Souza SFC, Ferraz CCR, et al. Effectiveness of 2% chlorhexidine gel and calcium hydroxide against Enterococcus faecalis in bovine root dentine in vitro. Int Endod J. 2003;36: 267–275.
  • 2. Portenier I, Waltimo TMT, Haapasalo M. Enterococcus faecalis: The root canal survivor and ‘star’ in post treatment disease. Endod Topics. 2003;6:135–159.
  • 3. Adl A, Hamedi S, Sedigh Shams M, Motamedifar M, Sobhnamayan F. The ability of triple antibiotic paste and calcium hydroxide in disinfection of dentinal tubules. Iran Endod J. 2014;9(2):123–126.
  • 4. Mozayeni MA, Haeri A, Dianat O, Jafari AR. Antimicrobial effects of four intracanal medicaments on Enterococcus faecalis: An in vitro study. Iran Endod J. 2014;9(3):195-198.
  • 5. Hoshino E, Takushige T. LSTR 3Mix-MP method: Better and efficient clinical procedure of lesion sterilization and tissue repair (LSTR) therapy. Dental Review. 1998;666:57–106.
  • 6. Arslan H, Çapar ID, Saygılı G, Uysal B, Gok T, Ertaş H. Efficacy of various irrigation protocols on the removal of triple antibiotic paste. Int Endod J. 2014;47(6) :594–599.
  • 7. Akçay M, Arslan H, Yasa B, Kavrık F, Yasa E. Spectrophotometric analysis of crown discoloration induced by various antibiotic pastes used in revascularization. J Endod. 2014;40(6):845–848.
  • 8. Horodniceanu T, Bougueleret L, El-Solh N, Bieth G, Delbos F. High-level, plasmid-borne resistance to gentamicin in Streptococcus faecalis subsp. zymogenes. Antimicrobial Agents Ch. 1979;16(5):686–689.
  • 9. Habib G, Hoen B, Tornos P, et al. ESC Committee for Practice Guidelines. Guidelines on the prevention, diagnosis, and treatment of infective endocarditis (new version 2009): The Task Force on the Prevention, Diagnosis, and Treatment of Infective Endocarditis of the European Society of Cardiology (ESC). Endorsed by the European Society of Clinical Microbiology and Infectious Diseases (ESCMID) and the International Society of Chemotherapy (ISC) for Infection and Cancer. Eur Heart J. 2009;30(19):2369–2413.
  • 10. Levay PF, Viljoen M. Lactoferrin: A general review. Haematologica. 1995;80:252–67
  • 11. Orsi N. The antimicrobial activity of lactoferrin: Current status and perspectives. Biometals. 2004;7(3):189–196.
  • 12. Velliyagounder K, Kaplan JB, Furgang D, et al. One of two human lactoferrin variants exhibit increased antibacterial and transcriptional activation activities and is associated with localized juvenile periodontitis. Infect Immun. 2003;71(11):6141–6147.
  • 13. Krupińska AM, Bogucki Z. Clinical aspects of the use of lactoferrin in dentistry. J Oral Biosci. 2021;Epub ahead of print. https://doi.org/10.1016/j.job.2021.02.005.
  • 14. Rosa L, Lepanto MS, Cutone A, Ianiro G, Pernarella S, Sangermano R, et al. Lactoferrin and oral pathologies: a therapeutic treatment. Biochem Cell Biol. 2021; 99(1): 81-90
  • 15. Villarreal JV, Jungfer C, Obst U, Schwartz T. DNase I and Proteinase K eliminate DNA from injured or dead bacteria but not from living bacteria in microbial reference systems and natural drinking water biofilms for subsequent molecular biology analyses. J Microbiol Methods. 2013;94(3):161–169.
  • 16. Lane DJ.16S/23S rRNA sequencing. In: Stackebrandt E, Goodfellow M. (Eds.), Nucleic acid techniques in Bacterial systematics. Wiley, Chichester, England. 1991.
  • 17. Schoch CL, Seifert KA, Huhndorf S, Robert V, Spouge JL, Levesque CA, et al. Fungal Barcoding Consortium; Fungal Barcoding Consortium Author List. Nuclear ribosomal internal transcribed spacer (ITS) region as a universal DNA barcode marker for Fungi. Proc Natl Acad Sci. 2012;109(16):6241-6246.
  • 18. Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and and the 2−ΔΔCT Method. Methods. 2001;25(4):402–408.
  • 19. Aksoy SÇ. Isolation of Enterococcus faecalis from permanent tooth root canals and identification by cultural and molecular methods and detection of antibiotic susceptibility (Master thesis). Ege University, Science Institute, Biology Department. 2008 Available from: http://acikerisim.ege.edu.tr:8081/xmlui/handle/11454/3402?show=full
  • 20. Siqueira JF Jr, Rôças IN. PCR methodology as a valuable tool for identification of endodontic pathogens. J Dent. 2003;31:333–339.
  • 21. Guerreiro JCM, Ochoa‐Rodrígez VM, Rodrigues EM, et al. Antibacterial activity, cytocompatibility and effect of Bio‐C Temp bioceramic intracanal medicament on osteoblast biology. Int Endod J. 2021; Epub ahead of print. https://doi.org/10.1111/iej.13502
  • 22. Song Y. PCR-based diagnostics for anaerobic infections. Anaerobe. 2005;11:79–91.
  • 23. Sheridan GEC, Szabo EA, Mackey BM. Effect of post-treatment holding conditions on detection of tufA mRNA in ethanol-treated Escherichia coli: implications for RT-PCR-based indirect viability tests. Lett Appl Microbiol. 1999;29:375–379.
  • 24. Trevors JT. Viable but non-culturable (VBNC) bacteria: Gene expression in planktonic and biofilm cells. J Microbiol Methods. 2011;86:266–273.
  • 25. Lleo M, Bonato B, Tafi MC. Adhesion to medical device materials and biofilm formation capability of some species of enterococci in different physiological states. FEMS Microbiol Lett. 2007;274(2):232-237.
  • 26. Chua EG, Parolia A, Ahlawat P, Pau A, Amalraj FD. Antifungal effectiveness of various intracanal medicaments against Candida albicans: An ex-vivo study. BMC Oral Health. 2014;14:53.
  • 27. Carreira C de M, dos Santos SS, Jorge AO, Lage-Marques JL. Antimicrobial effect of intracanal substances. J Appl Oral Sci. 2007;15(5):453–458.
  • 28. Gudipaneni, RK, Kumar RV, Jesudass G, Peddengatagari S, Duddu Y. Short-term comparative evaluation of antimicrobial efficacy of toothpaste containing lactoferrin, lysozyme, lactoperoxidase in children with severe early childhood caries: A clinical study. J Clin Diagnostic Res. 2014;8(4):18–20.
  • 29. Zheng JX, Wu Y, Lin ZW, Pu ZY, Yao WM, Chen Z, et al. Characteristics of and Virulence Factors Associated with Biofilm Formation in Clinical Enterococcus faecalis Isolates in China. Front Microbiol. 2017;8:2338.
  • 30. Seno Y, Kariyama R, Mitsuhata R, Monden K, Kumon H. Clinical implications of biofilm formation by Enterococcus faecalis in the urinary tract. Act Med Okayama. 2005;5:79–87.
  • 31. Sandoe JA, Witherden IR, Cove JH, Heritage J, Wilcox MH. Correlation between enterococcal biofilm formation in vitro and medical-device-related infection potential in vivo. J Med Microbiol. 2003;52:547–550.
  • 32. Mohammadi Z, Palazzi F, Giardino L. Microbial Biofilms in Endodontic Infections: An Update Review. Biomed J. 2013;36:59-70.
  • 33. Kristich CJ, Li YH, Cvitkovitch DG, Dunny GM. Esp-Independent Biofilm Formation by Enterococcus faecalis. J Bacteriol. 2004;186(1):154-163.
  • 34. Lleo MM, Tafi MC, Canepari P. Nonculturable Enterococcus faecalis cells are metabolically active and capable of resuming active growth. Syst Applied Microbiol. 1998;21(3):333–339.
  • 35. Shen Y, Stojicic S, Haapasalo M. Antimicrobial Efficacy of Chlorhexidine against Bacteria in Biofilms at Different Stages of Development. J Endod. 2011;37(5):657-661.
  • 36. Pinheiro ET, Gomes PBFA, Ferraz CCR, Teixeira FB, Zai AA, Souza-Filho FJ. Evaluation of root canal microorganisms isolated from teeth with endodontic failure and their antimicrobial susceptibility. Oral Microbiol Immun. 2003;18:100–103.
  • 37. Aslangul E, Ruimy R, Chau F, Garry L, Andremont A, Fantin B. Relationship between the level of acquired resistance to gentamicin and synergism with amoxicillin in Enterococcus faecalis. Antimicrob Agents Ch. 2005;49(10):4144-4148.
  • 38. Wang JT, Chang SC, Wang HY, Chen PC, Shiau YR, Lauderdale TL. TSAR Hospitals. High rates of multidrug resistance in Enterococcus faecalis and E. faecium isolated from inpatients and outpatients in Taiwan. Diagn Micr Infect Dis. 2013;75(4):406–411.
  • 39. Mizuhashi F, Koide K, Toya S, Takahashi M, Mizuhashi R, Shimomura H. Levels of the antimicrobial proteins lactoferrin and chromogranin in the saliva of individuals with oral dryness. J Prosthet Dent. 2015;113(1):35–38.
  • 40. Ammons MCB, Ward LS, James GA. Anti-biofilm efficacy of a lactoferrin/xylitol wound hydrogel used in combination with silver wound dressings. Int Wound J. 2011; 8:268–273.
  • 41. de Andrade FB, de Oliveira JC, Yoshie MT, Guimaraes BM, Gonçalves RB, Schwarcz WD. Antimicrobial activity and synergism of lactoferrin and lysozyme against cariogenic microorganisms. Braz Dent J. 2014,25(2):165–169.
  • 42. Alves FR, Silva MG, Rôças IN, Siqueira Jr. JF. Biofilm biomass disruption by natural substances with potential for endodontic use. Braz Oral Res. 2013;27(1):20-25.
  • 43. Cochrane S, Parashos P, Burrow MF. Effect on the mechanical properties of human and bovine dentine of intracanal medicaments and irrigants. Aust Dent J. 2019; 64:35–42.
  • 44. Yilmaz S, Dumani A, Yoldas O. The effect of antibiotic pastes on microhardness of dentin. Dent Traumatol. 2016;32(1):27-31.

Year 2021, Volume: 24 Issue: 3, 256 - 265, 15.09.2021

Burçin Arıcan , Hesna Sazak Öveçoğlu

https://doi.org/10.7126/cumudj.912285

Abstract

Project Number

SAG-C-DRP 150513–0154

References

  • 1. Gomes BPFA, Souza SFC, Ferraz CCR, et al. Effectiveness of 2% chlorhexidine gel and calcium hydroxide against Enterococcus faecalis in bovine root dentine in vitro. Int Endod J. 2003;36: 267–275.
  • 2. Portenier I, Waltimo TMT, Haapasalo M. Enterococcus faecalis: The root canal survivor and ‘star’ in post treatment disease. Endod Topics. 2003;6:135–159.
  • 3. Adl A, Hamedi S, Sedigh Shams M, Motamedifar M, Sobhnamayan F. The ability of triple antibiotic paste and calcium hydroxide in disinfection of dentinal tubules. Iran Endod J. 2014;9(2):123–126.
  • 4. Mozayeni MA, Haeri A, Dianat O, Jafari AR. Antimicrobial effects of four intracanal medicaments on Enterococcus faecalis: An in vitro study. Iran Endod J. 2014;9(3):195-198.
  • 5. Hoshino E, Takushige T. LSTR 3Mix-MP method: Better and efficient clinical procedure of lesion sterilization and tissue repair (LSTR) therapy. Dental Review. 1998;666:57–106.
  • 6. Arslan H, Çapar ID, Saygılı G, Uysal B, Gok T, Ertaş H. Efficacy of various irrigation protocols on the removal of triple antibiotic paste. Int Endod J. 2014;47(6) :594–599.
  • 7. Akçay M, Arslan H, Yasa B, Kavrık F, Yasa E. Spectrophotometric analysis of crown discoloration induced by various antibiotic pastes used in revascularization. J Endod. 2014;40(6):845–848.
  • 8. Horodniceanu T, Bougueleret L, El-Solh N, Bieth G, Delbos F. High-level, plasmid-borne resistance to gentamicin in Streptococcus faecalis subsp. zymogenes. Antimicrobial Agents Ch. 1979;16(5):686–689.
  • 9. Habib G, Hoen B, Tornos P, et al. ESC Committee for Practice Guidelines. Guidelines on the prevention, diagnosis, and treatment of infective endocarditis (new version 2009): The Task Force on the Prevention, Diagnosis, and Treatment of Infective Endocarditis of the European Society of Cardiology (ESC). Endorsed by the European Society of Clinical Microbiology and Infectious Diseases (ESCMID) and the International Society of Chemotherapy (ISC) for Infection and Cancer. Eur Heart J. 2009;30(19):2369–2413.
  • 10. Levay PF, Viljoen M. Lactoferrin: A general review. Haematologica. 1995;80:252–67
  • 11. Orsi N. The antimicrobial activity of lactoferrin: Current status and perspectives. Biometals. 2004;7(3):189–196.
  • 12. Velliyagounder K, Kaplan JB, Furgang D, et al. One of two human lactoferrin variants exhibit increased antibacterial and transcriptional activation activities and is associated with localized juvenile periodontitis. Infect Immun. 2003;71(11):6141–6147.
  • 13. Krupińska AM, Bogucki Z. Clinical aspects of the use of lactoferrin in dentistry. J Oral Biosci. 2021;Epub ahead of print. https://doi.org/10.1016/j.job.2021.02.005.
  • 14. Rosa L, Lepanto MS, Cutone A, Ianiro G, Pernarella S, Sangermano R, et al. Lactoferrin and oral pathologies: a therapeutic treatment. Biochem Cell Biol. 2021; 99(1): 81-90
  • 15. Villarreal JV, Jungfer C, Obst U, Schwartz T. DNase I and Proteinase K eliminate DNA from injured or dead bacteria but not from living bacteria in microbial reference systems and natural drinking water biofilms for subsequent molecular biology analyses. J Microbiol Methods. 2013;94(3):161–169.
  • 16. Lane DJ.16S/23S rRNA sequencing. In: Stackebrandt E, Goodfellow M. (Eds.), Nucleic acid techniques in Bacterial systematics. Wiley, Chichester, England. 1991.
  • 17. Schoch CL, Seifert KA, Huhndorf S, Robert V, Spouge JL, Levesque CA, et al. Fungal Barcoding Consortium; Fungal Barcoding Consortium Author List. Nuclear ribosomal internal transcribed spacer (ITS) region as a universal DNA barcode marker for Fungi. Proc Natl Acad Sci. 2012;109(16):6241-6246.
  • 18. Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and and the 2−ΔΔCT Method. Methods. 2001;25(4):402–408.
  • 19. Aksoy SÇ. Isolation of Enterococcus faecalis from permanent tooth root canals and identification by cultural and molecular methods and detection of antibiotic susceptibility (Master thesis). Ege University, Science Institute, Biology Department. 2008 Available from: http://acikerisim.ege.edu.tr:8081/xmlui/handle/11454/3402?show=full
  • 20. Siqueira JF Jr, Rôças IN. PCR methodology as a valuable tool for identification of endodontic pathogens. J Dent. 2003;31:333–339.
  • 21. Guerreiro JCM, Ochoa‐Rodrígez VM, Rodrigues EM, et al. Antibacterial activity, cytocompatibility and effect of Bio‐C Temp bioceramic intracanal medicament on osteoblast biology. Int Endod J. 2021; Epub ahead of print. https://doi.org/10.1111/iej.13502
  • 22. Song Y. PCR-based diagnostics for anaerobic infections. Anaerobe. 2005;11:79–91.
  • 23. Sheridan GEC, Szabo EA, Mackey BM. Effect of post-treatment holding conditions on detection of tufA mRNA in ethanol-treated Escherichia coli: implications for RT-PCR-based indirect viability tests. Lett Appl Microbiol. 1999;29:375–379.
  • 24. Trevors JT. Viable but non-culturable (VBNC) bacteria: Gene expression in planktonic and biofilm cells. J Microbiol Methods. 2011;86:266–273.
  • 25. Lleo M, Bonato B, Tafi MC. Adhesion to medical device materials and biofilm formation capability of some species of enterococci in different physiological states. FEMS Microbiol Lett. 2007;274(2):232-237.
  • 26. Chua EG, Parolia A, Ahlawat P, Pau A, Amalraj FD. Antifungal effectiveness of various intracanal medicaments against Candida albicans: An ex-vivo study. BMC Oral Health. 2014;14:53.
  • 27. Carreira C de M, dos Santos SS, Jorge AO, Lage-Marques JL. Antimicrobial effect of intracanal substances. J Appl Oral Sci. 2007;15(5):453–458.
  • 28. Gudipaneni, RK, Kumar RV, Jesudass G, Peddengatagari S, Duddu Y. Short-term comparative evaluation of antimicrobial efficacy of toothpaste containing lactoferrin, lysozyme, lactoperoxidase in children with severe early childhood caries: A clinical study. J Clin Diagnostic Res. 2014;8(4):18–20.
  • 29. Zheng JX, Wu Y, Lin ZW, Pu ZY, Yao WM, Chen Z, et al. Characteristics of and Virulence Factors Associated with Biofilm Formation in Clinical Enterococcus faecalis Isolates in China. Front Microbiol. 2017;8:2338.
  • 30. Seno Y, Kariyama R, Mitsuhata R, Monden K, Kumon H. Clinical implications of biofilm formation by Enterococcus faecalis in the urinary tract. Act Med Okayama. 2005;5:79–87.
  • 31. Sandoe JA, Witherden IR, Cove JH, Heritage J, Wilcox MH. Correlation between enterococcal biofilm formation in vitro and medical-device-related infection potential in vivo. J Med Microbiol. 2003;52:547–550.
  • 32. Mohammadi Z, Palazzi F, Giardino L. Microbial Biofilms in Endodontic Infections: An Update Review. Biomed J. 2013;36:59-70.
  • 33. Kristich CJ, Li YH, Cvitkovitch DG, Dunny GM. Esp-Independent Biofilm Formation by Enterococcus faecalis. J Bacteriol. 2004;186(1):154-163.
  • 34. Lleo MM, Tafi MC, Canepari P. Nonculturable Enterococcus faecalis cells are metabolically active and capable of resuming active growth. Syst Applied Microbiol. 1998;21(3):333–339.
  • 35. Shen Y, Stojicic S, Haapasalo M. Antimicrobial Efficacy of Chlorhexidine against Bacteria in Biofilms at Different Stages of Development. J Endod. 2011;37(5):657-661.
  • 36. Pinheiro ET, Gomes PBFA, Ferraz CCR, Teixeira FB, Zai AA, Souza-Filho FJ. Evaluation of root canal microorganisms isolated from teeth with endodontic failure and their antimicrobial susceptibility. Oral Microbiol Immun. 2003;18:100–103.
  • 37. Aslangul E, Ruimy R, Chau F, Garry L, Andremont A, Fantin B. Relationship between the level of acquired resistance to gentamicin and synergism with amoxicillin in Enterococcus faecalis. Antimicrob Agents Ch. 2005;49(10):4144-4148.
  • 38. Wang JT, Chang SC, Wang HY, Chen PC, Shiau YR, Lauderdale TL. TSAR Hospitals. High rates of multidrug resistance in Enterococcus faecalis and E. faecium isolated from inpatients and outpatients in Taiwan. Diagn Micr Infect Dis. 2013;75(4):406–411.
  • 39. Mizuhashi F, Koide K, Toya S, Takahashi M, Mizuhashi R, Shimomura H. Levels of the antimicrobial proteins lactoferrin and chromogranin in the saliva of individuals with oral dryness. J Prosthet Dent. 2015;113(1):35–38.
  • 40. Ammons MCB, Ward LS, James GA. Anti-biofilm efficacy of a lactoferrin/xylitol wound hydrogel used in combination with silver wound dressings. Int Wound J. 2011; 8:268–273.
  • 41. de Andrade FB, de Oliveira JC, Yoshie MT, Guimaraes BM, Gonçalves RB, Schwarcz WD. Antimicrobial activity and synergism of lactoferrin and lysozyme against cariogenic microorganisms. Braz Dent J. 2014,25(2):165–169.
  • 42. Alves FR, Silva MG, Rôças IN, Siqueira Jr. JF. Biofilm biomass disruption by natural substances with potential for endodontic use. Braz Oral Res. 2013;27(1):20-25.
  • 43. Cochrane S, Parashos P, Burrow MF. Effect on the mechanical properties of human and bovine dentine of intracanal medicaments and irrigants. Aust Dent J. 2019; 64:35–42.
  • 44. Yilmaz S, Dumani A, Yoldas O. The effect of antibiotic pastes on microhardness of dentin. Dent Traumatol. 2016;32(1):27-31.
There are 44 citations in total.

Details

Primary Language English
Subjects Health Care Administration
Journal Section Original Research Articles
Authors

Burçin Arıcan 0000-0001-5757-0571

Hesna Sazak Öveçoğlu 0000-0003-4709-422X

Project Number SAG-C-DRP 150513–0154
Publication Date September 15, 2021
Submission Date April 9, 2021
Published in Issue Year 2021Volume: 24 Issue: 3

Cite

EndNote Arıcan B, Sazak Öveçoğlu H (September 1, 2021) IN VITRO COMPARISON OF THE ANTIMICROBIAL EFFECTS OF DIFFERENT ROOT CANAL MEDICAMENTS ON ENTEROCOCCUS FAECALIS AND CANDIDA ALBICANS. Cumhuriyet Dental Journal 24 3 256–265.
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