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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
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.
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.
