Acta Scientific Dental Sciences (ISSN: 2581-4893)

Research ArticleVolume 5 Issue 6

The Effect of Warm Gutta-Percha Techniques on Viability and Mineralized Tissue Associated Gene’s Transcripts of Human Periodontal-Ligament Mesenchymal Stem-Cells

Nazife Tuğba Azmaz1, Serife Buket Bozkurt2, Sema Sezgin Hakki3 and Sema Belli4*

1Specialist Endodontist, Meram Research and Training Hospital, Konya, Turkey

2Research Lab of Faculty of Dentistry, Hacettepe University, Ankara, Turkey

3Full-Time Professor in Department of Periodontology, Selcuk University, Konya, Turkey

4Full-Time Professor in Department of Endodontics, Selcuk University, Konya, Turkey

*Corresponding Author: Sema Belli, Full-Time Professor in Department of Endodontics, Selcuk University, Konya, Turkey.

Received: March 20, 2021; Published: : May 05, 2021

Citation: Sema Belli., et al. “The Effect of Warm Gutta-Percha Techniques on Viability and Mineralized Tissue Associated Gene’s Transcripts of Human Periodontal-Ligament Mesenchymal Stem-Cells". Acta Scientific Dental Sciences 5.6 (2021): 11-20.


Aim: The purpose of the this study was to assess the effect of continuous wave of condensation (CWCT) and thermoplastic gutta-percha injection (TGI) techniques on the viability and mRNA expressions of mineralized tissue-associated proteins of human periodontal-ligament mesenchymal stem-cells (hPDL-MSCs).

Methods: hPDL-MSCs were isolated from premolar-teeth. Roots of human premolar-teeth were placed vertically to the cell-culture dishes. Six groups were designed as follows: control 1 (without teeth, C1); control 2 (with teeth C2); AH Plus group (AH); single-cone obturation group (SC); CWCT and TGI. The viability of the hPDL-MSCs was analyzed by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide at 24 h and 96h. RNA was isolated from the hPDL-MSCs at 24 hrs and cDNA synthesis was performed. The mRNA expression of mineralized tissue-markers including bone sialoprotein (BSP), osteocalcin (OCN), runt-related transcription factor-2 (Runx2), type I collagen (COL I), alkaline phosphatase (ALP), were evaluated by quantitative real time polymerase chain reaction (qRT-PCR).

Results: Reduced cell viability was observed in all groups except the controls. When SC and CWCT and TGI groups were compared, it was observed that heat had a significant negative effect on cell viability (p < 0.05). Reduction in the mRNA expressions of BSP, OCN, and ALP were observed at the test groups and this negative effect was higher in the heat treated groups (p < 0.05).

Conclusion: Warm gutta-percha techniques reduced mRNA expressions of mineralized tissue-associated proteins which are critical for the functions of PDL-MSCs during healing at the apical region after endodontic treatment.

Keywords: Associated Proteins; Mineralized Tissue; Periodontal Ligament; Mesenchymal Stem Cells; Warm Gutta-Percha Techniques


  1. LLI Grossman., et al. “Endodontic practice”. Lea and Febiger (1988).
  2. P Dummer. “Comparison of undergraduate endodontic teaching programmes in the United Kingdom and in some dental schools in Europe and the United States”. International Endodontic Journal 4 (1991): 169-177.
  3. L Spångberg. “Endodontic treatment of teeth without apical periodontitis”. Essential Endodontics. Blackwell, London (1998): 211-241.
  4. J Marciano and PM Michailesco. “Dental gutta-percha: chemical composition, X-ray identification, enthalpic studies, and clinical implications”. Journal of Endodontics 4 (1989): 149-153.
  5. V Vishwanath and HM Rao. “Gutta-percha in endodontics-A comprehensive review of material science”. Journal of Conservative Dentistry JCD 3 (2019): 216.
  6. G Hommez., et al. “Endodontic treatment performed by Flemish dentists. Part 2. Canal filling and decision making for referrals and treatment of apical periodontitis”. International Endodontic Journal 5 (2003): 344-351.
  7. R Krug., et al. “Technical quality of a matching-taper single-cone filling technique following rotary instrumentation compared with lateral compaction after manual preparation: a retrospective study”. Clinical Oral Investigations 2 (2017): 643-652.
  8. H Schilder. “Filling root canals in three dimensions”. Journal of Endodontics 4 (2006): 281-290.
  9. RA Barkhordar., et al. “Evaluation of temperature rise on the outer surface of teeth during root canal obturation techniques”. Quintessence International 7 (1990): 585-588.
  10. JJ Jurcak., et al. “In vitro intracanal temperatures produced during warm lateral condensation of gutta-percha”. Journal of Endodontics 1 (1992): 1-3.
  11. R Weller and K Koch. “In vitro radicular temperatures produced by injectable thermoplasticized gutta percha”. International Endodontic Journal 2 (1995):
  12. A Eriksson and T Albrektsson. “Temperature threshold levels for heat-induced bone tissue injury: a vital-microscopic study in the rabbit”. Journal of Prosthetic Dentistry 1 (1983): 101-107.
  13. E Saunders. “In vivo findings associated with heat generation during thermomechanical compactionof gutta‐percha. Part II. Histological response to temperature elevation on the external surface of the root”. International Endodontic Journal 5 (1990): 268-274.
  14. FS Lee., et al. “A comparison of root surface temperatures using different obturation heat sources”. Journal of Endodontics 9 (1998): 617-620.
  15. O Trubiani., et al. “Periodontal ligament stem cells: current knowledge and future perspectives”. Stem Cells and Development 15 (2019): 995-1003.
  16. HJ Kim., et al. “Cytotoxicity and genotoxicity of newly developed calcium phosphate-based root canal sealers”. Korean Medical Journal Information 1 (2006): 36-49.
  17. SS Hakki., et al. “Effects of mineral trioxide aggregate on cell survival, gene expression associated with mineralized tissues, and biomineralization of cementoblasts”. Journal of Endodontics 4 (2009): 513-519.
  18. SS Hakki., et al. “Comparison of mesenchymal stem cells isolated from pulp and periodontal ligament”. Journal of Periodontology 2 (2015): 283-291.
  19. E Saunders and W Saunders. “The heat generated on the external root surface during post space preparation”. International Endodontic Journal 4 (1989): 169-173.
  20. C Hauman and R Love. “Biocompatibility of dental materials used in contemporary endodontic therapy: a review. Part 1. Intracanal drugs and substances”. International Endodontic Journal 2 (2003): 75-85.
  21. G Botton., et al. “Toxicity of irrigating solutions and pharmacological associations used in pulpectomy of primary teeth”. International Endodontic Journal8 (2016): 746-754.
  22. R Shinagawa-Ohama., et al. “Heterogeneous human periodontal ligament-committed progenitor and stem cell populations exhibit a unique cementogenic property under in vitro and in vivo conditions”. Stem Cells and Development 9 (2017): 632-645.
  23. SS Hakki., et al. “Osteogenic differentiation of MC3T3-E1 cells on different titanium surfaces”. Biomedical Materials 4 (2012): 045006.
  24. NT Azmaz., et al. “Warm Gutta-Percha Techniques Regulate Cell Viability, Heat Shock, and Mineralized Tissue–associated Proteins of Cementoblasts”. Journal of Endodontics 7 (2020): 957-963.
  25. M Dominici., et al. “Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement”. Cytotherapy4 (2006): 315-317.
  26. KJ Livak and TD Schmittgen. “Analysis of relative gene expression data using real-time quantitative PCR and the 2− ΔΔCT method”. Methods4 (2001): 402-408.
  27. W Zhang., et al. “Evaluation of cytotoxicity of MTAD using the MTT-tetrazolium method”. Journal of Endodontics 10 (2003): 654-657.
  28. T Mosmann. “Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays”. The Journal of Immunological Methods 1-2 (1983): 55-63.
  29. SS Hakki., et al. “Bone morphogenetic protein -2, -6, and -7 differently regulate osteogenic differentiation of human periodontal ligament stem cells”. Journal of Biomedical Materials Research Part B: Applied 1 (2014): 119-130.
  30. WA Castelli., et al. “Periodontium response to a root canal condensing device (Endotec)”. Oral Surgery, Oral Medicine, Oral Pathology, and Oral Radiology 3 (1991): 333-337.
  31. EM Hardie. “Further studies on heat generation during obturation techniques involving thermally softened gutta-percha”. International Endodontic Journal 3 (1987): 122-127.
  32. JL Gutmann., et al. “Evaluation of heat transfer during root canal obturation with thermoplasticized gutta-percha. Part II. In vivo response to heat levels generated”. Journal of Endodontics 9 (1987): 441-448.
  33. AD Romero., et al. “Heat transfer to the periodontal ligament during root obturation procedures using an in vitro model”. Journal of Endodontics 2 (2000): 85-87.
  34. SAT Al-Shimari and NE Al-Nuaimi. “Elevation in surface temperature of root canals obturated with different thermoplasticized gutta-percha obturation techniques-an in vitro study”. Journal of Baghdad College of Dentistry 1 (2014): 67-70.
  35. SS Hakki., et al. “Periodontal ligament fibroblast response to root perforations restored with different materials–a laboratory study”. International Endodontic Journal 3 (2012): 240-248.
  36. SS Hakki., et al. “The response of cementoblasts to calcium phosphate resin–based and calcium silicate–based commercial sealers”. International Endodontic Journal 3 (2013): 242-252.
  37. YJ Choi., et al. “Enhanced osteogenesis by collagen-binding peptide from bone sialoprotein in vitro and in vivo”. Journal of Biomedical Materials Research Part A 2 (2013): 547-554.
  38. JA D'Errico., et al. “Expression of bone associated markers by tooth root lining cells, in situ and in vitro”. Bone 2 (1997): 117-126.
  39. SS Hakki., et al. “Bone sialoprotein gene transfer to periodontal ligament cells may not be sufficient to promote mineralization in vitro or in vivo”. Journal of Periodontology 2 (2006): 167-173.
  40. JB Lian., et al. “Osteocalcin gene promoter: unlocking the secrets for regulation of osteoblast growth and differentiation”. Journal of Cellular Biochemistry S30‒31 (1998): 62-72.
  41. AL Bronckers., et al. “Studies of osteocalcin function in dentin formation in rodent teeth”. The European Journal of Oral Sciences 3 (1998): 795-807.
  42. C Liu and J Sun. “Hydrolyzed tilapia fish collagen induces osteogenic differentiation of human periodontal ligament cells”. Biomedical Materials 6 (2015): 065020.
  43. A Pilloni., et al. “Migliaccio Analysis of human alveolar osteoblast behavior on a nano-hydroxyapatite substrate: an in vitro study”. BMC Oral Health1 (2014): 14-22.
  44. A Neve., et al. “Osteocalcin: skeletal and extra‐skeletal effects”. Journal of Cellular Physiology 6 (2013): 1149-1153.
  45. R Miron and Y Zhang. “Osteoinduction: a review of old concepts with new standards”. Journal of Dental Research 8 (2012): 736-744.
  46. C Shui and A Scutt. “Mild heat shock induces proliferation, alkaline phosphatase activity, and mineralization in human bone marrow stromal cells and Mg-63 cells in vitro”. Journal of Bone and Mineral Research 4 (2001): 731-741.

Copyright: © 2021 Sema Belli., et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

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