Cytotoxicity Evaluation of Minimum Antibacterial Values of Different Medicaments Used in Endodontic Regenerative ProceduresFunding This study was supported by Tehran University of Medical Sciences (grant no: 35692).
31 December 2019 (online)
Objectives This study aimed to evaluate the cytotoxicity of minimum antibacterial values of medicaments used in endodontic regeneration on stem cells.
Materials and Methods “Minimum inhibitory concentration,” “minimum bactericidal concentration,” and “minimum biofilm inhibitory concentration” of triple and double antibiotic paste, a modified triple antibiotic paste (minocycline replaced by clindamycin), Augmentin, and calcium hydroxide were determined using Enterococcus faecalis (ATCC 29212) by microtiter plate method. Direct cytotoxic effects of drugs were evaluated by lactate dehydrogenase and water-soluble tetrazolium salt-1 assays using stem cells of apical papilla obtained from immature third molars via enzymatic digestion.
Statistical Analysis Data were analyzed using IBM SPSS Statistics 24, one-way analysis of variance and post hoc comparisons. The statistical power was set at p < 0.05.
Results All medicaments caused similar cytotoxicity and cell proliferation at “minimum inhibitory concentration” (p > 0.05) except Augmentin which was significantly more toxic than others (p < 0.05). At “minimum bactericidal concentration,” calcium hydroxide was more toxic than other drugs (p < 0.001), but its adverse effect on cell proliferation was the same as Augmentin (p > 0.05). Triple and double antibiotic paste revealed similar favorable effects in terms of toxicity and proliferation rate at most of the tested concentrations (p > 0.05). At “minimum biofilm inhibitory concentration” both the modified paste and Augmentin caused less proliferation rate than triple and double antibiotic paste (p < 0.001and p < 0.05, respectively) and Augmentin induced more cytotoxicity (p < 0.05).
Conclusions Considering the antimicrobial potency, triple antibiotic paste seems to be the safest drug for the stem cells of apical papilla, while Augmentin may have some adverse effects.
- 1 Hargreaves KM, Diogenes A, Teixeira FB. Treatment options: biological basis of regenerative endodontic procedures. J Endod 2013; 39 (03) (Suppl): S30-S43
- 2 Jeeruphan T, Jantarat J, Yanpiset K, Suwannapan L, Khewsawai P, Hargreaves KM. Mahidol study 1: comparison of radiographic and survival outcomes of immature teeth treated with either regenerative endodontic or apexification methods: a retrospective study. J Endod 2012; 38 (10) 1330-1336
- 3 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. J Endod 2011; 37 (02) 133-138
- 4 Lin LM, Shimizu E, Gibbs JL, Loghin S, Ricucci D. Histologic and histobacteriologic observations of failed revascularization/revitalization therapy: a case report. J Endod 2014; 40 (02) 291-295
- 5 Diogenes A, Ruparel NB, Shiloah Y, Hargreaves KM. Regenerative endodontics: a way forward. J Am Dent Assoc 2016; 147 (05) 372-380
- 6 Chen MY, Chen KL, Chen CA, Tayebaty F, Rosenberg PA, Lin LM. Responses of immature permanent teeth with infected necrotic pulp tissue and apical periodontitis/abscess to revascularization procedures. Int Endod J 2012; 45 (03) 294-305
- 7 Chueh LH, Huang GT. Immature teeth with periradicular periodontitis or abscess undergoing apexogenesis: a paradigm shift. J Endod 2006; 32 (12) 1205-1213
- 8 Nosrat A, Seifi A, Asgary S. Regenerative endodontic treatment (revascularization) for necrotic immature permanent molars: a review and report of two cases with a new biomaterial. J Endod 2011; 37 (04) 562-567
- 9 Trope M. Treatment of the immature tooth with a non-vital pulp and apical periodontitis. Dent Clin North Am 2010; 54 (02) 313-324
- 10 McTigue DJ, Subramanian K, Kumar A. Case series: management of immature permanent teeth with pulpal necrosis: a case series. Pediatr Dent 2013; 35 (01) 55-60
- 11 Nosrat A, Li KL, Vir K, Hicks ML, Fouad AF. Is pulp regeneration necessary for root maturation?. J Endod 2013; 39 (10) 1291-1295
- 12 Ruparel NB, Teixeira FB, Ferraz CC, Diogenes A. Direct effect of intracanal medicaments on survival of stem cells of the apical papilla. J Endod 2012; 38 (10) 1372-1375
- 13 Kim JH, Kim Y, Shin SJ, Park JW, Jung IY. Tooth discoloration of immature permanent incisor associated with triple antibiotic therapy: a case report. J Endod 2010; 36 (06) 1086-1091
- 14 Althumairy RI, Teixeira FB, Diogenes A. Effect of dentin conditioning with intracanal medicaments on survival of stem cells of apical papilla. J Endod 2014; 40 (04) 521-525
- 15 Labban N, Yassen GH, Windsor LJ, Platt JA. The direct cytotoxic effects of medicaments used in endodontic regeneration on human dental pulp cells. Dent Traumatol 2014; 30 (06) 429-434
- 16 Diogenes AR, Ruparel NB, Teixeira FB, Hargreaves KM. Translational science in disinfection for regenerative endodontics. J Endod 2014; 40 (04) (Suppl): S52-S57
- 17 AlSaeed T, Nosrat A, Melo MA. et al. Antibacterial efficacy and discoloration potential of endodontic topical antibiotics. J Endod 2018; 44 (07) 1110-1114
- 18 Sabrah AH, Yassen GH, Gregory RL. Effectiveness of antibiotic medicaments against biofilm formation of Enterococcus faecalis and Porphyromonas gingivalis. J Endod 2013; 39 (11) 1385-1389
- 19 Trevino EG, Patwardhan AN, Henry MA. 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 (08) 1109-1115
- 20 CLSI. Performance Standards for Antimicrobial Susceptibility Testing. 26th ed. CLSI supplement M100S. Wayne, PA: Clinical and Laboratory Standards Institute 2016
- 21 Andrews JM. Determination of minimum inhibitory concentrations. J Antimicrob Chemother 2001; 48 (01) Suppl 1 5-16
- 22 Xu X, Zhou XD, Wu CD. The tea catechin epigallocatechin gallate suppresses cariogenic virulence factors of Streptococcus mutans. Antimicrob Agents Chemother 2011; 55 (03) 1229-1236
- 23 Fouad AF. The microbial challenge to pulp regeneration. Adv Dent Res 2011; 23 (03) 285-289
- 24 Sabrah AH, Yassen GH, Liu WC, Goebel WS, Gregory RL, Platt JA. The effect of diluted triple and double antibiotic pastes on dental pulp stem cells and established Enterococcus faecalis biofilm. Clin Oral Investig 2015; 19 (08) 2059-2066
- 25 Chuensombat S, Khemaleelakul S, Chattipakorn S, Srisuwan T. Cytotoxic effects and antibacterial efficacy of a 3-antibiotic combination: an in vitro study. J Endod 2013; 39 (06) 813-819
- 26 Kamocki K, Nör JE, Bottino MC. Effects of ciprofloxacin-containing antimicrobial scaffolds on dental pulp stem cell viability-In vitro studies. Arch Oral Biol 2015; 60 (08) 1131-1137
- 27 Karczewski A, Feitosa SA, Hamer EI. et al. Clindamycin-modified triple antibiotic nanofibers: a stain-free antimicrobial intracanal drug delivery system. J Endod 2018; 44 (01) 155-162
- 28 Albuquerque MT, Evans JD, Gregory RL, Valera MC, Bottino MC. Antibacterial TAP-mimic electrospun polymer scaffold: effects on P. gingivalis-infected dentin biofilm. Clin Oral Investig 2016; 20 (02) 387-393
- 29 Galluzzi L, Aaronson SA, Abrams J. et al. Guidelines for the use and interpretation of assays for monitoring cell death in higher eukaryotes. Cell Death Differ 2009; 16 (08) 1093-1107
- 30 Peters OA. Research that matters - biocompatibility and cytotoxicity screening. Int Endod J 2013; 46 (03) 195-197
- 31 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 (07) 667-675
- 32 Hosseini Matin M, Zare Jahromi M, Fesharaki M, Ostad Sharif M. Cytotoxicity of triple antibiotic paste and calcium hydroxide against cultured human dental pulp fibroblasts. J Dental School Shahid Beheshti Univ Med Sci 2015; 33 (03) 196-204