Antimicrobial efficacy of a new tri-antibiotic combination against resistant endodontic pathogens: An in-vitro study

Authors

  • Prasanna T. Dahake DMIMS (Deemed to be University), Sawangi (M), Wardha, Maharashtra, India. Pin code – 442004. https://orcid.org/0000-0003-0295-5751
  • Sudhindra M Baliga Department of Pedodontics and Preventive Dentistry, Sharad Pawar Dental College and Hospital, DMIMS (Deemed to be University), Sawangi (M), Wardha, Maharashtra, India. Pin code – 442004. https://orcid.org/0000-0002-4799-8205

DOI:

https://doi.org/10.14295/bds.2021.v24i1.2186

Abstract

Background: Removal of all the pathogenic bacteria from the root canal system is of prime importance for the success of endodontic therapy. Objective: The study aimed to determine the antimicrobial efficacy of three antibiotics and their new combination against selected endodontic pathogens. Methods: In this in-vitro study, we used bacterial strains associated with the refractory endodontic condition and determined MIC and MBC of Clindamycin (C), Metronidazole (M), Doxycycline (D) as well as their combination CMD. We cultured Candida Albicans, Pseudomonas Aeruginosa, Escherichia Coli, Enterococcus Faecalis, Streptococcus Mutans, Bacillus Subtilis subsp. spizizenii, Actinomyces Actinomycetemcomitans on selective culture media. We analyzed the data using paired 't' test, one-way ANOVA, and Tuckey's HSD post hoc test. Results: Clindamycin inhibited the growth of C. Albicans (90%) and S. Mutans (90%) significantly and P. Aeruginosa, E. Coli, E. Faecalis, B. Subtilis, and A. Actinomycetemcomitans were resistant to it. Metronidazole did not inhibit any of the bacteria. Doxycycline inhibited C. Albicans (90%), P. Aeruginosa (90%), and S. Mutans (90%) significantly while E. Coli, E. Faecalis, B. Subtilis, and A. Actinomycetemcomitans were resistant to it. The combination of CMD inhibited all the microbes significantly. However, at bactericidal concentrations of CMD, E. Faecalis (p = 0.024), B. Subtilis (p = 0.021) and A. Actinomycetemcomitans (p = 0.041) were eliminated significantly, while C. Albicans (p = 0.164), P. Aeruginosa (p = 0.489), E. Coli (p = 0.106) and S. Mutans (p = 0.121) showed resistance. Conclusion: Combination CMD can be used against resistant endodontic pathogens to achieve predictable endodontic results.

KEYWORDS

Antimicrobial agents; Clindamycin; Doxycycline; Metronidazole; Root canal therapy.    

Author Biographies

Prasanna T. Dahake, DMIMS (Deemed to be University), Sawangi (M), Wardha, Maharashtra, India. Pin code – 442004.

Ph. D. Student,

Pharmacology and Pedodontics,

(Interdisciplinary Sciences),

DMIMS (Deemed to be University),

Sawangi (M), Wardha, Maharashtra, India.

Pin code – 442004.

Ph: +919096027989,                

E-mail:prasannadahake@gmail.com

Sudhindra M Baliga, Department of Pedodontics and Preventive Dentistry, Sharad Pawar Dental College and Hospital, DMIMS (Deemed to be University), Sawangi (M), Wardha, Maharashtra, India. Pin code – 442004.

MDS, Ph.D. Professor,

Department of Pedodontics and Preventive Dentistry,

Sharad Pawar Dental College and Hospital,

DMIMS (Deemed to be University), Sawangi (M), Wardha,

Maharashtra, India. Pin code – 442004.

Ph: +919960640066,

E‑mail: baligams@hotmail.com

References

Casadevall A, Pirofski L. Host-pathogen interactions: the attributes of virulence. J Infect Dis. 2001 Aug 1;184(3):337-44. doi: 10.1086/322044.

Koo H, Allan RN, Howlin RP, Stoodley P, Hall-Stoodley L. Targeting microbial biofilms: current and prospective therapeutic strategies. Nat Rev Microbiol. 2017 Dec;15(12):740-55. doi: 10.1038/nrmicro.2017.99.

Gabrilska RA, Rumbaugh KP. Biofilm models of polymicrobial infection. Future Microbiol. 2015;10(12):1997-2015. doi: 10.2217/fmb.15.109.

Peterson SN, Snesrud E, Liu J, Ong AC, Kilian M, Schork NJ, et al. The dental plaque microbiome in health and disease. PLoS One. 2013;8(3):e58487. doi: 10.1371/journal.pone.0058487.

van Houte J, Lopman J, Kent R. The predominant cultivable flora of sound and carious human root surfaces. J Dent Res. 1994 Nov;73(11):1727-34. doi: 10.1177/00220345940730110801.

Ghabraei S, Bolhari B, Sabbagh MM, Afshar MS. Comparison of antimicrobial effects of triple antibiotic paste and calcium hydroxide mixed with 2% chlorhexidine as intracanal medicaments against enterococcus faecalis biofilm. J Dent (Tehran). 2018 May;15(3):151-60.

Gilad JZ, Teles R, Goodson M, White RR, Stashenko P. Development of a clindamycin-impregnated fiber as an intracanal medication in endodontic therapy. J Endod. 1999 Nov;25(11):722-7. doi: 10.1016/S0099-2399(99)80117-4.

Neelakantan P, Romero M, Vera J, Daood U, Khan AU, Yan A, et al. Biofilms in endodontics-current status and future directions. Int J Mol Sci. 2017 Aug 11;18(8):1748. doi: 10.3390/ijms18081748.

Yoo YJ, Perinpanayagam H, Oh S, Kim AR, Han SH, Kum KY. Endodontic biofilms: contemporary and future treatment options. Restor Dent Endod. 2019 Jan 31;44(1):e7. doi: 10.5395/rde.2019.44.e7.

Panpaliya NP, Dahake PT, Kale YJ, Dadpe MV, Kendre SB, Siddiqi AG, et al. In vitro evaluation of antimicrobial property of silver nanoparticles and chlorhexidine against five different oral pathogenic bacteria. Saudi Dent J. 2019 Jan;31(1):76-83. doi: 10.1016/j.sdentj.2018.10.004.

Qaiyumi S. Macro and microdilution methods of antimicrobial susceptibility testing. In: Schwabe R, Steele-Moore L, Goodwil AC. (Eds.), Antimicrobial Susceptibility Testing Protocols. CRC Press, United State of America. 2007: pp 75-9.

Espinel-Ingroff A, Colombo AL, Cordoba S, Dufresne PJ, Fuller J, Ghannoum M, et al. International evaluation of mic distributions and epidemiological cutoff value (ecv) definitions for fusarium species identified by molecular methods for the clsi broth microdilution method. Antimicrob Agents Chemother. 2015 Dec 7;60(2):1079-84. doi: 10.1128/AAC.02456-15.

Segura-Egea JJ, Gould K, Şen BH, Jonasson P, Cotti E, Mazzoni A, et al. Antibiotics in Endodontics: a review. Int Endod J. 2017 Dec;50(12):1169-84. doi: 10.1111/iej.12741.

Liljemark WF, Bloomquist C. Human oral microbial ecology and dental caries and periodontal diseases. Crit Rev Oral Biol Med. 1996;7(2):180-98. doi: 10.1177/10454411960070020601.

Sundqvist G, Figdor D, Persson S, Sjögren U. Microbiologic analysis of teeth with failed endodontic treatment and the outcome of conservative re-treatment. Oral Surg Oral Med Oral Pathol Oral Radiol Endod. 1998 Jan;85(1):86-93. doi: 10.1016/s1079-2104(98)90404-8.

Radcliffe CE, Potouridou L, Qureshi R, Habahbeh N, Qualtrough A, Worthington H, et al. Antimicrobial activity of varying concentrations of sodium hypochlorite on the endodontic microorganisms Actinomyces israelii, A. naeslundii, Candida albicans and Enterococcus faecalis. Int Endod J. 2004 Jul;37(7):438-46. doi: 10.1111/j.1365-2591.2004.00752.x.

Molander A, Reit C, Dahlén G, Kvist T. Microbiological status of root-filled teeth with apical periodontitis. Int Endod J. 1998 Jan;31(1):1-7.

Nair PN, Sjögren U, Krey G, Kahnberg KE, Sundqvist G. Intraradicular bacteria and fungi in root-filled, asymptomatic human teeth with therapy-resistant periapical lesions: a long-term light and electron microscopic follow-up study. J Endod. 1990 Dec;16(12):580-8. doi: 10.1016/S0099-2399(07)80201-9.

Yamane K, Ogawa K, Yoshida M, Hayashi H, Nakamura T, Yamanaka T, et al. Identification and characterization of clinically isolated biofilm-forming gram-positive rods from teeth associated with persistent apical periodontitis. J Endod. 2009 Mar;35(3):347-52. doi: 10.1016/j.joen.2008.11.032.

Evans M, Davies JK, Sundqvist G, Figdor D. Mechanisms involved in the resistance of Enterococcus faecalis to calcium hydroxide. Int Endod J. 2002 Mar;35(3):221-8. doi: 10.1046/j.1365-2591.2002.00504.x.

Pereira RS, Rodrigues VAA, Furtado WT, Gueiros S, Pereira GS, Avila-Campos MJ. Microbial analysis of root canal and periradicular lesion associated to teeth with endodontic failure. Anaerobe. 2017 Dec;48:12-18. doi: 10.1016/j.anaerobe.2017.06.016.

Garcez AS, Ribeiro MS, Tegos GP, Núñez SC, Jorge AO, Hamblin MR. Antimicrobial photodynamic therapy combined with conventional endodontic treatment to eliminate root canal biofilm infection. Lasers Surg Med. 2007 Jan;39(1):59-66. doi: 10.1002/lsm.20415.

Nomura R, Ogaya Y, Nakano K. Contribution of the collagen-binding proteins of streptococcus mutans to bacterial colonization of inflamed dental pulp. PLoS One. 2016 Jul 21;11(7):e0159613. doi: 10.1371/journal.pone.0159613.

Del Fabbro M, Corbella S, Sequeira-Byron P, et al. Endodontic procedures for retreatment of periapical lesions. Cochrane Database Syst Rev. 2016;10(10):CD005511. Published 2016 Oct 19. doi:10.1002/14651858.

Mohammadi Z, Abbott PV. On the local applications of antibiotics and antibiotic-based agents in endodontics and dental traumatology. Int Endod J. 2009 Jul;42(7):555-67. doi: 10.1111/j.1365-2591.2009.01564.x.

Sandor GK, Low DE, Judd PL, Davidson RJ. Antimicrobial treatment options in the management of odontogenic infections. J Can Dent Assoc. 1998 Jul-Aug;64(7):508-14.

Cvek M, Cleaton-Jones P, Austin J, Lownie J, Kling M, Fatti P. Effect of topical application of doxycycline on pulp revascularization and periodontal healing in reimplanted monkey incisors. Endod Dent Traumatol. 1990 Aug;6(4):170-6.

Torabinejad M, Khademi AA, Babagoli J, Cho Y, Johnson WB, Bozhilov K, et al. A new solution for the removal of the smear layer. J Endod. 2003 Mar;29(3):170-5. doi: 10.1097/00004770-200303000-00002.

Chan Y, Chan CH. Antibiotic resistance of pathogenic bacteria from odontogenic infections in Taiwan. J Microbiol Immunol Infect. 2003 Jun;36(2):105-10.

Smieja M. Current indications for the use of clindamycin: A critical review. Can J Infect Dis. 1998 Jan;9(1):22-8. doi: 10.1155/1998/538090.

Behra-Miellet J, Dubreuil L, Jumas-Bilak E. Antianaerobic activity of moxifloxacin compared with that of ofloxacin, ciprofloxacin, clindamycin, metronidazole and beta-lactams. Int J Antimicrob Agents. 2002 Nov;20(5):366-74. doi: 10.1016/s0924-8579(02)00209-1.

Downloads

Published

2020-09-30

Issue

Section

Clinical or Laboratorial Research Manuscript