Advertisement

Clinical Oral Investigations

, Volume 23, Issue 3, pp 1051–1059 | Cite as

The antimicrobial properties, cytotoxicity, and differentiation potential of double antibiotic intracanal medicaments loaded into hydrogel system

  • Patrick W. McIntyre
  • Jennifer L. Wu
  • Rasika Kolte
  • Ru Zhang
  • Richard L. Gregory
  • Angela Bruzzaniti
  • Ghaeth H. YassenEmail author
Original Article

Abstract

Objective

This study evaluated the antimicrobial properties, cytotoxicity, and mineralization potential of methylcellulose hydrogels loaded with low concentrations of double antibiotic pastes (DAP).

Materials and methods

The direct and residual antibacterial effects of 1, 5, and 10 mg/mL of DAP loaded into hydrogels as well as calcium hydroxide (Ca(OH)2) were tested against single-species biofilms of Enterococcus faecalis and dual-species biofilms (Enterococcus faecalis and Prevotella intermedia). The effects of DAP hydrogels on proliferation and mineralization of dental pulp stem cells (DPSC) were tested using MTT assays, alkaline phosphate activity (ALP), and alizarin red staining. Fisher’s exact tests, Wilcoxon rank sum tests, and one-way ANOVA were used for statistical analyses (α = 0.05).

Results

All tested concentrations of DAP hydrogels as well as Ca(OH)2 demonstrated significant direct antibacterial effects against single- and dual-species biofilms. However, only 5 and 10 mg/mL of DAP hydrogels exhibited significant residual antibacterial effects against both types of tested biofilms. Only 1 mg/mL of DAP hydrogels did not have significant negative effects on DPSC viability, ALP activity, and mineralization nodule formation. However, 5 and 10 mg/mL of DAP hydrogels caused significant negative effects on cytotoxicity and mineralization nodule formation of DPSC.

Conclusions

Hydrogels containing 1 mg/mL DAP offered significant direct antibacterial effects against single- and dual-species biofilms without causing significant negative effects on viability, ALP activity, and mineralization nodule formation of DPSC.

Clinical relevance

The methylcellulose-based hydrogel proposed in this study can be used clinically as a biocompatible system to deliver controlled low concentrations of DAP.

Keywords

Antimicrobials Cytotoxicity Double antibiotic paste Hydrogels Mineralization 

Notes

Funding

This study was supported by Indiana University School of Dentistry

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

All procedures performed in studies involving human teeth were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards.

Informed consent

Informed consent was obtained from all individual participants included in the study.

References

  1. 1.
    Lin J, Zeng Q, Wei X, Zhao W, Cui M, Gu J, Lu J, Yang M, Ling J (2017) Regenerative endodontics versus Apexification in immature permanent teeth with apical periodontitis: a prospective randomized controlled study. J Endod 43:1821–1827CrossRefGoogle Scholar
  2. 2.
    Jiang X, Liu H, Peng C (2017) Clinical and radiographic assessment of the efficacy of a collagen membrane in regenerative endodontics: a randomized, controlled clinical trial. J Endod 43:1465–1471CrossRefGoogle Scholar
  3. 3.
    Fouad AF (2017) Microbial factors and antimicrobial strategies in dental pulp regeneration. J Endod 43:S46–s50CrossRefGoogle Scholar
  4. 4.
    Diogenes AR, Ruparel NB, Teixeira FB, Hargreaves KM (2014) Translational science in disinfection for regenerative endodontics. J Endod 40:S52–S57CrossRefGoogle Scholar
  5. 5.
    Trevino EG, Patwardhan AN, Henry MA, Perry G, Dybdal-Hargreaves N, Hargreaves KM, Diogenes A (2011) 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 37:1109–1115CrossRefGoogle Scholar
  6. 6.
    Althumairy RI, Teixeira FB, Diogenes A (2014) Effect of dentin conditioning with intracanal medicaments on survival of stem cells of apical papilla. J Endod 40:521–525CrossRefGoogle Scholar
  7. 7.
    American Association of Endodontists (2017) AAE Clinical Considerations for a Regenerative Procedure. Available at: https://www.aae.org/uploadedfiles/publications_and_research/research/currentregenerativeendodonticconsiderations.pdf Accessed october 20, 2017
  8. 8.
    Ruparel NB, Teixeira FB, Ferraz CC et al (2012) Direct effect of intracanal medicaments on survival of stem cells of the apical papilla. J Endod 38:1372–1375CrossRefGoogle Scholar
  9. 9.
    Tagelsir A, Yassen GH, Gomez GF, Gregory RL (2016) Effect of antimicrobials used in regenerative endodontic procedures on 3-week-old enterococcus faecalis biofilm. J Endod 42:258–262CrossRefGoogle Scholar
  10. 10.
    Latham J, Fong H, Jewett A, Johnson JD, Paranjpe A (2016) Disinfection efficacy of current regenerative endodontic protocols in simulated necrotic immature permanent teeth. J Endod 42:1218–1225CrossRefGoogle Scholar
  11. 11.
    Jenks DB, Ehrlich Y, Spolnik K, Gregory RL, Yassen GH (2016) Residual antibiofilm effects of various concentrations of double antibiotic paste used during regenerative endodontics after different application times. Arch Oral Biol 70:88–93CrossRefGoogle Scholar
  12. 12.
    Sabrah AH, Yassen GH, Liu WC et al (2015) The effect of diluted triple and double antibiotic pastes on dental pulp stem cells and established enterococcus faecalis biofilm. Clin Oral Investig 19:2059–2066CrossRefGoogle Scholar
  13. 13.
    Alyas SM, Fischer BI, Ehrlich Y, Spolnik K, Gregory RL, Yassen GH (2016) Direct and indirect antibacterial effects of various concentrations of triple antibiotic pastes loaded in a methylcellulose system. J Oral Sci 58:575–582CrossRefGoogle Scholar
  14. 14.
    Labban N, Yassen GH, Windsor LJ, Platt JA (2014) The direct cytotoxic effects of medicaments used in endodontic regeneration on human dental pulp cells. Dent Traumatol 30:429–434CrossRefGoogle Scholar
  15. 15.
    Alghilan MA, Windsor LJ, Palasuk J, Yassen GH (2017) Attachment and proliferation of dental pulp stem cells on dentine treated with different regenerative endodontic protocols. Int Endod J 50:667–675CrossRefGoogle Scholar
  16. 16.
    Kim KW, Yassen GH, Ehrlich Y, Spolnik K, Platt JA, Windsor LJ (2015) The effects of radicular dentine treated with double antibiotic paste and ethylenediaminetetraacetic acid on the attachment and proliferation of dental pulp stem cells. Dent Traumatol 31:374–379CrossRefGoogle Scholar
  17. 17.
    Kaushik SN, Scoffield J, Andukuri A, Alexander GC, Walker T, Kim S, Choi SC, Brott BC, Eleazer PD, Lee JY, Wu H, Childers NK, Jun HW, Park JH, Cheon K (2015) Evaluation of ciprofloxacin and metronidazole encapsulated biomimetic nanomatrix gel on enterococcus faecalis and Treponema denticola. Biomater Res 19:9CrossRefGoogle Scholar
  18. 18.
    Palasuk J, Kamocki K, Hippenmeyer L, Platt JA, Spolnik KJ, Gregory RL, Bottino MC (2014) Bimix antimicrobial scaffolds for regenerative endodontics. J Endod 40:1879–1884CrossRefGoogle Scholar
  19. 19.
    Pankajakshan D, Albuquerque MT, Evans JD et al (2016) Triple antibiotic polymer nanofibers for Intracanal drug delivery: effects on dual species biofilm and cell function. J Endod 42:1490–1495CrossRefGoogle Scholar
  20. 20.
    Jacobs JC, Troxel A, Ehrlich Y, Spolnik K, Bringas JS, Gregory RL, Yassen GH (2017) Antibacterial effects of antimicrobials used in regenerative endodontics against biofilm Bacteria obtained from mature and immature teeth with necrotic pulps. J Endod 43:575–579CrossRefGoogle Scholar
  21. 21.
    Sabrah AH, Yassen GH, Spolnik KJ et al (2015) Evaluation of residual antibacterial effect of human radicular dentin treated with triple and double antibiotic pastes. J Endod 41:1081–1084CrossRefGoogle Scholar
  22. 22.
    Ray HL Jr, Marcelino J, Braga R, Horwat R, Lisien M, Khaliq S (2016) Long-term follow up of revascularization using platelet-rich fibrin. Dent Traumatol 32:80–84CrossRefGoogle Scholar
  23. 23.
    Maniglia-Ferreira C, de Almeida Gomes F, Vitoriano MM (2017) Intentional replantation of an avulsed immature permanent incisor: a case report. J Endod 43:1383–1386CrossRefGoogle Scholar
  24. 24.
    Akcay M, Arslan H, Yasa B, Kavrık F, Yasa E (2014) Spectrophotometric analysis of crown discoloration induced by various antibiotic pastes used in revascularization. J Endod 40:845–848CrossRefGoogle Scholar
  25. 25.
    Fouad AF, Nosrat A (2013) Pulp regeneration in previously infected root canal space. Endod Top 28:24–37CrossRefGoogle Scholar
  26. 26.
    Verma P, Nosrat A, Kim JR, Price JB, Wang P, Bair E, Xu HH, Fouad AF (2017) Effect of residual Bacteria on the outcome of pulp regeneration in vivo. J Dent Res 96:100–106CrossRefGoogle Scholar
  27. 27.
    Xie Q, Johnson BR, Wenckus CS, Fayad MI, Wu CD (2012) Efficacy of berberine, an antimicrobial plant alkaloid, as an endodontic irrigant against a mixed-culture biofilm in an in vitro tooth model. J Endod 38:1114–1117CrossRefGoogle Scholar
  28. 28.
    Nagata JY, Soares AJ, Souza-Filho FJ, Zaia AA, Ferraz CCR, Almeida JFA, Gomes BPFA (2014) Microbial evaluation of traumatized teeth treated with triple antibiotic paste or calcium hydroxide with 2% chlorhexidine gel in pulp revascularization. J Endod 40:778–783CrossRefGoogle Scholar
  29. 29.
    Bakopoulou A, Leyhausen G, Volk J, Tsiftsoglou A, Garefis P, Koidis P, Geurtsen W (2011) Comparative analysis of in vitro osteo/odontogenic differentiation potential of human dental pulp stem cells (DPSCs) and stem cells from the apical papilla (SCAP). Arch Oral Biol 56:709–721CrossRefGoogle Scholar
  30. 30.
    Abuarqoub D, Awidi A, Abuharfeil N (2015) Comparison of osteo/odontogenic differentiation of human adult dental pulp stem cells and stem cells from apical papilla in the presence of platelet lysate. Arch Oral Biol 60:1545–1553CrossRefGoogle Scholar
  31. 31.
    Sonoyama W, Liu Y, Yamaza T, Tuan RS, Wang S, Shi S, Huang GTJ (2008) Characterization of the apical papilla and its residing stem cells from human immature permanent teeth: a pilot study. J Endod 34:166–171CrossRefGoogle Scholar
  32. 32.
    Janvikul W, Uppanan P, Thavornyutikarn B, Prateepasen R, Swasdison S (2007) Fibroblast interaction with carboxymethylchitosan-based hydrogels. J Mater Sci Mater Med 18:943–949CrossRefGoogle Scholar
  33. 33.
    Phumpatrakom P, Srisuwan T (2014) Regenerative capacity of human dental pulp and apical papilla cells after treatment with a 3-antibiotic mixture. J Endod 40:399–405CrossRefGoogle Scholar
  34. 34.
    Chen L, Zheng L, Jiang J, Gui J, Zhang L, Huang Y, Chen X, Ji J, Fan Y (2016) Calcium hydroxide-induced proliferation, migration, osteogenic differentiation, and mineralization via the mitogen-activated protein kinase pathway in human dental pulp stem cells. J Endod 42:1355–1361CrossRefGoogle Scholar
  35. 35.
    Narita H, Itoh S, Imazato S, Yoshitake F, Ebisu S (2010) An explanation of the mineralization mechanism in osteoblasts induced by calcium hydroxide. Acta Biomater 6:586–590CrossRefGoogle Scholar
  36. 36.
    Nosrat A, Homayounfar N, Oloomi K (2012) Drawbacks and unfavorable outcomes of regenerative endodontic treatments of necrotic immature teeth: a literature review and report of a case. J Endod 38:1428–1434CrossRefGoogle Scholar
  37. 37.
    Shah N, Logani A, Bbaskar U et al (2008) Efficacy of revascularization to induce apexification/apexogensis in infected, nonvital, immature teeth: a pilot clinical study. J Endod 34:919–925CrossRefGoogle Scholar
  38. 38.
    Song M, Cao Y, Shin SJ, Shon WJ, Chugal N, Kim RH, Kim E, Kang MK (2017) Revascularization-associated Intracanal calcification: assessment of prevalence and contributing factors. J Endod 43:2025–2033CrossRefGoogle Scholar
  39. 39.
    Galler KM, Krastl G, Simon S, van Gorp G, Meschi N, Vahedi B, Lambrechts P (2016) European Society of Endodontology position statement: revitalization procedures. Int Endod J 49:717–723CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  • Patrick W. McIntyre
    • 1
  • Jennifer L. Wu
    • 1
  • Rasika Kolte
    • 1
  • Ru Zhang
    • 1
    • 2
  • Richard L. Gregory
    • 1
  • Angela Bruzzaniti
    • 1
  • Ghaeth H. Yassen
    • 1
    • 3
    Email author
  1. 1.Department of Biomedical and Applied SciencesIndiana University School of DentistryIndianapolisUSA
  2. 2.Department of Endodontics, Beijing Stomatological HospitalCapital Medical UniversityBeijingChina
  3. 3.Department of EndodonticsCase School of Dental MedicineClevelandUSA

Personalised recommendations