Document Type : Original Article

Authors

1 Pedodontist, Dept. of Pediatric Dentistry, Dental School, Hamedan University of Medical Sciences, Hamedan, Iran.

2 Dentist, Resident of Pediatric Dentistry, Dental School, Hamedan University of Medical Sciences, Hamedan, Iran.

3 Dept. of Anatomy, School of Medicine, Hamadan University of Medical Sciences, Hamadan, Iran.

4 Dept. of Biostatistics, Hamadan University of Medical Sciences, Hamadan, Iran.

Abstract

Statement of the Problem: The use of a new antimicrobial combination in the regenerative endodontic treatment of immature teeth pulp necrosis is a well-known method. Concerns have been raised about the destructive effect of this combination on the stem cells from the apical papilla of permanent human teeth, and there is a study gap.
Purpose: The main objective of the present study was to investigate the cytotoxic effect of modified triple antibiotic paste (mTAP) on stem cells from the apical papilla (SCAPs) of permanent human teeth.
Materials and Method: In this in vitro study, stem cells were removed from the immature teeth. After cultivation and third passage, metronidazole, ciprofloxacin, minocycline, and clindamycin were placed in the cell culture medium alone ,  paired, and in combinations as triple antibiotic paste (TAP) (metronidazole, ciprofloxacin, and minocycline) and mTAP (metronidazole, ciprofloxacin, clindamycin) with doses of 25, 50, 100, 200, 400μg/ml. After 1 and 3 days, cell viability in the culture medium was assessed using the MTT method ([4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide). SPSS software version 24, descriptive statistics methods, and statistical tests such as Kruskal-Wallis and Mann-Whitney tests were adopted to analyze the data.
Results: Analysis of MTT findings indicated that the use of mTAP at 100μg/ml and TAP at 200μg/ml had no adverse cytotoxic effect on stem cells in the first 24 hours, compared to the control group. The cell viability decreased at higher concentrations, although it was not statistically significant. After 72 hours, the toxicity of concentrations higher than 100μg/ml of mTAP and 400 μg/ml of TAP significantly mitigated the percentage of viable cells.
Conclusion: The obtained results demonstrated that the concentration of 100 μg/ml of mTAP could replace TAP in regenerative endodontic treatments at the studied time intervals without worrying about the toxicity.

Keywords

  • Phumpatrakom P, Srisuwan T. Regenerative capacity of human dental pulp and apical papilla cells after treatment with a 3-antibiotic Endod. 2014; 40: 399-405.
  • Banchs F, Trope M. Revascularization of immature permanent teeth with apical periodontitis: new treatment protocol? Endod. 2004; 30: 196-200.
  • Lovelace TW, Henry MA, Hargreaves KM, Diogenes A. Evalu­ation of the delivery of mesenchymal stem cells into the root canal space of necrotic immature teeth after clinical regenera­tive endodontic procedure. Endod. 2011; 37: 133-138.
  • Hoshino E, Kurihara-Ando N, Sato I, Uematsu H, Sato M, Kota K, et al. In vitro antibacterial susceptibility of bacteria taken from infected root dentine to a mixture of ciprofloxacin, metronidazole and minocycline. Int Endod. 1996; 29: 125-130.
  • Segura-Egea JJ, Velasco-Ortega E, Torres-Lagares D, Velasco-Ponferrada MC, Monsalve-Guil L, Llamas-Carreras JM. Pattern of antibiotic prescription in the management of endodontic infections amongst Spanish oral surgeons. Int Endod J. 2010; 43: 342-350.
  • Sato I, Ando-Kurihara N, Kota K, Iwaku M, Hoshino E. Sterilization of infected rootcanal dentine by topical application of a mixture of ciprofloxacin, metronidazole and minocycline in situ. Int Endod J. 1996; 29: 118–124.
  • Kim H, Kim Y, Shin S, Park W, Ung IY. Tooth discoloration of immature permanent incisor associated with triple antibiotic therapy: a case report. Endod. 2010; 36: 1086–1091.
  • Reynolds K, Ohnson D, Cohenca N. Pulp revascularization of nerotic bilateral bicuspids using a modified novel technique to eliminate potential coronal discolouration: a case report. Int Endod. 2009; 42: 84–92.
  • Kim B, Song MJ, Shin SJ, Park JW. Prevention of tooth discoloration associated with triple antibiotics. Restor Dent Endod. 2012; 37: 119–122
  • Zargar N, Rayat*Hosein*Abadi M, Sabeti M, Yadegari Z, Akbarzadeh*Baghban A, Dianat O. Antimicrobial efficacy of clindamycin and triple antibiotic paste as root canal medicaments on tubular infection: an in vitro study. Australian Endo J. 2019; 45: 86-91.
  • Casamassimo PS, Fields Jr HW, McTigue DJ, Nowak A .Pediatric dentistry: infancy through adolescence. 5th ed. China: Linda Duncan; 2013. p.500-505.
  • Rafatjou R, Yousefimashouf R, Farhadian M, Afzalsoltani S. Evaluation of the Antimicrobial Efficacy of two Combinations of drugs on bacteria taken from infected primary teeth: An in vitro Study. J Eur Arch Paed Dent. 2019; 1: 1-7.
  • Lin J, Zeng Q, Wei X, Zhao W, Cui M, Gu J, et al. Regenerative endodontics versus apexification in immature permanent teeth with apical periodontitis: a prospective randomized controlled study. J Endo. 2017; 43: 1821-1827.
  • Chuensombat S, Khemaleelakul S, Chattipakorn S, Srisuwan T. Cytotoxic effects and antibacterial efficacy of a 3-antibiotic combination: an in vitro study. J Endo. 2013; 39: 813-819.
  • Rafatou R, Amiri I, Aneshin A. Effect of Calcium-enriched Mixture (CEM) cement on increasing mineralization in stem cells from the dental pulps of human exfoliated deciduous teeth. J Dent Res Dent Clin Dent Pros. 2018; 12: 233.
  • Masoum T, Amiri I, Rafatou R. Effect of chitosan on osteogenic properties of mesenchymal stem cell of exfoliated deciduous teeth. J Dent Med. 2013; 26: 55-63.
  • Ruparel NB, Teixeira FB, Ferraz CC, Diogenes A. Direct effect of intracanal medicaments on survival of stem cells of the apical papilla. Endod. 2012; 38: 1372-1375.
  • Saberi EA, Karkehabadi H, Mollashahi NF. Cytotoxicity of Various Endodontic Materials on Stem Cells of Human Apical Papilla. Iranian Endo J. 2016; 11: 17-22.
  • Phumpatrakom P, Srisuwan T. Regenerative capacity of human dental pulp and apical papilla cells after treatment with a 3-antibiotic mixture. Endod. 2014; 40: 399-405.
  • Kitikuson P, Srisuwan T. Attachment ability of human apical papilla cells to root dentin surfaces treated with either 3Mix or calcium hydroxide. Endod. 2016; 42:89-94.
  • Banchs F, Trope M. Revascularization of immature permanent teeth with apical periodontitis: new treatment protocol? Endod. 2004; 30: 196-200.
  • Diogenes A, Ruparel NB, Shiloah Y, Hargreaves KM. Regenerative endodontics: a way forward. J Am Dent Assoc. 2016; 147: 372–380.
  • Chan EK, Desmeules M, Cielecki M, Dabbagh B, dos Santos BF. Longitudinal cohort study of regenerative endodontic treatment for immature necrotic permanent teeth. J Endo. 2017; 43: 395-400.
  • Karczewski A, Feitosa SA, Hamer EI, Pankajakshan D, Gregory RL, Spolnik KJ, et al. Clindamycin-modified Triple Antibiotic Nanofibers: A Stain-free Antimicrobial Intracanal Drug Delivery System. J Endod. 2018; 44: 155-162. 
  • Bi J, Liu Y, Liu XM, Jiang LM, Chen X. iRoot FM exerts an antibacterial effect on Porphyromonas endodontalis and improves the properties of stem cells from the apical papilla. Int Endod J. 2018; 51: 1139-1148.
  • Trevino EG, Patwardhan AN, Henry MA, Perry G, Dybdal-Hargreaves N, Hargreaves KM, et al. Effect of irrigants on the survival of human stem cells of the apical papilla in a platelet-rich plasma scaffold in human root tips. J Endod. 2011; 37: 1109-1115.
  • Khoshkhounejad M, Sobhi*Afshar M, Jabalameli F, Emaneini M, Sharifian M. Cytotoxicity Evaluation of Minimum Antibacterial Values of Different Medicaments Used in Endodontic Regenerative Procedures. Eur J Dent. 2019; 13: 514-520.
  • Althumairy RI, Teixeira FB, Diogenes A. Effect of dentin condi­tioning with intracanal medicaments on survival of stem cells of apical papilla. Endod. 2014; 40: 521–525.
  • Labban N, Yassen GH, Windsor L, Platt A. The direct cytotoxic effects of medicaments used in endodontic regeneration on human dental pulp cells. Dent Traumatol. 2014; 30: 429–434.
  • Sabrah AH, Yassen GH, Liu WC, Goebel WS, Gregory RL, Platt A. The effect of diluted triple and double antibiotic pastes on dental pulp stem cells and established Enterococcus faecalis biofilm. Clin Oral Investig. 2015; 19: 2059–2066.
  • Alghilan MA, Windsor LJ, Palasuk J, Yassen GH. Attachment and proliferation of dental pulp stem cells on dentine treated with different regenerative endodontic protocols. Int Endod J. 2017; 50: 667-675. 
  • Kamocki K, Nor E, Bottino MC. Effects of ciprofloxacin-con­taining antimicrobial scaffolds on dental pulp stem cell viabil­ity in vitro studies. Arch Oral Biol. 2015; 60: 1131–1137.
  • Albuquerque MT, Evans D, Gregory RL, Valera MC, Bottino MC. Antibacterial TAP-mimic electrospun polymer scaffold: effects on P. gingivalis-infected dentin biofilm. Clin Oral Investig. 2016; 20: 387–393.