# EDTA enhances cementum-like tissue formation, TGF-β1 & VEGF in rat molars during regenerative endodontics

**Authors:** Alexandre Henrique dos REIS-PRADO, Isabela Joane Prado SILVA, Juliana GOTO, Nathália Evelyn da Silva MACHADO, Gleide Fernandes de AVELAR, Juliano Douglas Silva ALBERGARIA, Raphael Escorsim SZAWKA, Marco Cícero BOTTINO, Luciano Tavares Angelo CINTRA, Edilson ERVOLINO, Antônio Paulino RIBEIRO-SOBRINHO, Francine BENETTI

PMC · DOI: 10.1590/1807-3107bor-2025.vol39.068 · Brazilian Oral Research · 2025-07-07

## TL;DR

Using EDTA during dental procedures in rats improved tissue formation and increased certain growth factors in the root area.

## Contribution

This study shows that EDTA irrigation enhances cementum-like tissue and specific growth factors during regenerative endodontics in rats.

## Key findings

- EDTA irrigation increased cementum-like tissue formation compared to NaOCl alone.
- TGF-β1 and VEGF immunolabeling were enhanced in the apical region with EDTA.
- No significant difference in FGF-2 immunolabeling was observed between groups.

## Abstract

This study investigated the influence of ethylenediaminetetraacetic acid (EDTA) irrigation on cementum-like tissue formation and TGF-β1, FGF-2, and VEGF immunolabeling during regenerative endodontic procedures (REPs) in immature rat molars. The lower first molars of 12 four-week-old male rats (80 g) underwent pulpectomy in the mesial canal and were randomly categorized into two experimental groups (n = 6): sodium hypochlorite (NaOCl) – irrigated for 5 min with 2.5% NaOCl; and NaOCl-EDTA – irrigated with 2.5% NaOCl, followed by 5 min of 17% EDTA. After inducing bleeding with a size 10 K-file, the cavities were sealed. Untreated molars served as control-15d (n = 3) and immediate control (n = 3). Either immediately or after 15 days, the animals were euthanized, and the teeth were collected for histomorphometric and immunohistochemical (TGF-β1, FGF-2, and VEGF) analysis. The results were analyzed by the Mann-Whitney U-test (p < 0.05). Histomorphometric analysis revealed increased cementum-like tissue formation in the NaOCl-EDTA group compared with that in the NaOCl group (p < 0.05). Regarding growth factor immunolabeling, the NaOCl-EDTA group exhibited enhanced TGF-β1 and VEGF immunolabeling in the root tip area and the center region of the apical third of the pulp tissue, compared with that in the NaOCl group (p < 0.05); however, no significant difference was observed in FGF-2 (p > 0.05). In conclusion, the use of EDTA in REPs positively affected the formation of cementum-like tissue and TGF-β1 and VEGF in the apical region but did not influence FGF-2.

## Linked entities

- **Proteins:** TGFB1 (transforming growth factor beta 1), VEGFA (vascular endothelial growth factor A), FGF2 (fibroblast growth factor 2)
- **Chemicals:** EDTA (PubChem CID 6049), sodium hypochlorite (PubChem CID 23665760), NaOCl (PubChem CID 23665760)
- **Species:** Mus musculus (taxon 10090)

## Full-text entities

- **Genes:** Fgf2 (fibroblast growth factor 2) [NCBI Gene 54250] {aka Fgf-2, Fgf2a, bFGF}, Vegfa (vascular endothelial growth factor A) [NCBI Gene 83785] {aka VEGF-A, VEGF111, VEGF164, VPF, Vegf}, Tgfb1 (transforming growth factor, beta 1) [NCBI Gene 59086] {aka Tgfb}
- **Diseases:** bleeding (MESH:D006470)
- **Chemicals:** NaOCl (MESH:D012973), EDTA (MESH:D004492)
- **Species:** Rattus norvegicus (brown rat, species) [taxon 10116]

## Full text

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## Figures

2 figures with captions in the complete paper: https://tomesphere.com/paper/PMC12237413/full.md

## References

40 references — full list in the complete paper: https://tomesphere.com/paper/PMC12237413/full.md

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Source: https://tomesphere.com/paper/PMC12237413