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Related Experiment Video

Updated: Jun 13, 2026

Isolation of Mesenchymal Stem Cells from Human Alveolar Periosteum and Effects of Vitamin D on Osteogenic Activity of Periosteum-derived Cells
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Isolation of Mesenchymal Stem Cells from Human Alveolar Periosteum and Effects of Vitamin D on Osteogenic Activity of Periosteum-derived Cells

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Effect of Adjunctive Ozone Application Protocols on Dentin-Derived Growth Factor Release: An In Vitro Study.

Sude Göbüt1,2, Melis Oya Ateş1, Ali Keleş1

  • 1Department of Endodontics, Faculty of Dentistry, Bolu Abant Izzet Baysal University, Bolu 14030, Türkiye.

Journal of Clinical Medicine
|June 12, 2026
PubMed
Summary

Adjunctive ozone application protocols influence dentin-derived transforming growth factor beta 1 (TGF-β1) release in regenerative endodontic treatment (RET). Ozonated ethylenediaminetetraacetic acid (EDTA) followed by ozone gas yielded the highest TGF-β1 levels, while insulin-like growth factor 1 (IGF-1) remained unaffected.

Keywords:
EDTAgrowth factor releasemicro-CTozoneregenerative endodontics

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Area of Science:

  • Biomaterials Science
  • Endodontics
  • Regenerative Medicine

Background:

  • Regenerative endodontic treatment (RET) relies on dentin-derived bioactive molecules.
  • Ethylenediaminetetraacetic acid (EDTA) is commonly used for dentin conditioning to promote molecule release.
  • The impact of adjunctive ozone therapy on growth factor release in RET is not fully understood.

Purpose of the Study:

  • To evaluate the effects of two adjunctive ozone application protocols combined with chelation on dentin-derived transforming growth factor beta 1 (TGF-β1) and insulin-like growth factor 1 (IGF-1) release.
  • To compare the measurable release of TGF-β1 and IGF-1 under different ozone and EDTA application strategies.

Main Methods:

  • Sixty-four human premolars were divided into four groups: EDTA alone, EDTA with ozonated water/gas, ozonated EDTA with gas, and a negative control.
  • Intracanal dentin surfaces were exposed, and specimens were incubated in phosphate-buffered saline (PBS) for 1 and 7 days.
  • TGF-β1 and IGF-1 levels were quantified using ELISA and normalized to dentin surface area measured by micro-CT.

Main Results:

  • No growth factors were detected in the negative control group.
  • Adjunctive ozone protocols significantly affected TGF-β1 release by day 7, with ozonated EDTA followed by ozone gas showing approximately threefold higher levels compared to EDTA with ozonated water/gas.
  • No significant differences in IGF-1 levels were observed between groups or over time.

Conclusions:

  • Adjunctive ozone application demonstrates a protocol-dependent influence on dentin-derived TGF-β1 release, with the ozonated EDTA followed by ozone gas protocol yielding the highest values.
  • IGF-1 release was not significantly affected by the tested ozone protocols.
  • These in vitro findings highlight measurable growth factor release but do not confirm in situ activation or clinical regenerative efficacy.