Effect of ethylenediaminetetraacetic acid irrigation on immune‐inflammatory response in teeth submitted to regenerative endodontic therapy

Abstract

Aim To analyse longitudinally the immune‐inflammatory response in teeth of mice that underwent a regenerative protocol with or without the use of ethylenediaminetetraacetic acid (EDTA) to irrigate the root canal system. Methodology First maxillary molars of mice were devitalized using size 10 and 15 files. Teeth were divided into the following groups: Empty – the canals were left empty; Blood Clot (BC) – the canals were filled with a blood clot; and EDTA + Blood – the canals were irrigated with 0.06 mL of 17% EDTA for 1 min and filled with a blood clot. Access cavities were restored with Coltosol®. Animals were sacrificed at 7, 14 or 21 days after the operative procedures, and teeth were collected. RNA was extracted, mRNA expression of the cytokines IGF, NGF, IL‐1α, IL‐10, TGF and VEGF was assessed using real‐time PCR, and the anova Kruskal–Wallis test was used. Results IL‐1 mRNA expression was significantly higher in the EDTA + BC group than in the Empty and BC groups at the 7th and 14th days of evaluation (P < 0.05). IL‐10 mRNA expression was similar across the three groups at all time periods. TGF‐β mRNA expression in the EDTA + BC group was significantly higher on the 7th and 21st days than on the 14th (P < 0.05); at day 21, TGF‐β mRNA expression was similar between the BC and EDTA + BC groups but significantly higher than in the Empty group (P < 0.05). IGF mRNA expression was significantly higher in the EDTA + BC group than in the other groups at all time periods. VEGF mRNA expression remained unchanged throughout the experimental period in all groups (P > 0.05). NGF mRNA expression was similar amongst all groups at the 7th and 21st days (P > 0.05). At the 14th day, however, there was a significant increase in NGF mRNA expression in the EDTA + Blood group (P < 0.05) when compared with the expression in the other groups. Conclusion EDTA promoted increased expression of factors that have the potential to improve the outcome of regenerative endodontic treatment.

Influence of ethylenediaminetetraacetic acid on regenerative endodontics: A systematic review

“…For the in vivo category, four animal studies were evaluated (Bracks et al, 2019; El Ashry et al, 2016; Kawamura et al, 2016; Yamauchi et al, 2011). Two studies only used 17% EDTA as the irrigating solution (Bracks et al, 2019; Yamauchi et al, 2011), whilst other studies also used guanidine hydrochloride (GdnHCl; Kawamura et al, 2016) and mixture tetracycline citric acid and detergent (MTAD; El Ashry et al, 2016). Regarding the regenerative protocols, three studies performed pulpectomies associated or not with an induction of periapical lesion in canine (El Ashry et al, 2016; Yamauchi et al, 2011) and mice posterior teeth (Bracks et al, 2019), whilst one used a subcutaneous tooth roots transplant into immunodeficient mice comparing an EDTA‐treated tooth to an untreated control (Kawamura et al, 2016).…”

“…The expression of signalling molecules following dentine demineralization might modulate cellular activity from the periapical area (Gonçalves et al, 2016; Taweewattanapaisan et al, 2019), thus playing a crucial role in the intracanal tissues’ neoformation (Bracks et al, 2019). A total of 16 studies evaluated the release of endogenous growth factors from dentine using different irrigating protocols with dentine discs.…”

“…Whereas TGF‐β may have immunosuppressive effects against the production of pro‐inflammatory cytokines (Maciel et al, 2012), the conditioning of the dentine with EDTA during REP did not influence tissue inflammation (El Ashry et al, 2016). In other analysis, one study found a high expression of IL‐1 in the group of EDTA‐treated dentine using real‐time PCR (Bracks et al, 2019). These controversial results may have taken place due to differences in the methods of analysis, since the presence of pro‐inflammatory cytokine does not exactly depend on the presence of inflammatory cells (Benetti et al, 2018).…”

“…TGF‐β1 is one of the most important stimulating molecules for the recruitment, proliferation and differentiation of stem cells (Bracks et al, 2019; Galler et al, 2015; Gonçalves et al, 2016). Besides enhancing odontoblastic differentiation and reparative dentinogenesis (Kucukkaya Eren et al, 2021), TGF‐β1 also shows substantial anti‐inflammatory effects by regulating pro‐inflammatory cytokines (Bracks et al, 2019). Moreover, the vascular formation after intracanal bleeding plays a critical role in cellular activity (Galler et al, 2015).…”

Successful Revascularization Procedure in an Immature Permanent Necrotic Second Lower Molar: A Case Report With 4.5-YEARS Follow-Up

“…Many biologically active growth factors trapped in the dentin matrix during the dentinogenesis, capable of promoting the survival, proliferation, and differentiation of DPSCs, are solubilized by EDTA, increasing their bioavailability. 42,43 Also, EDTA prevents the deleterious effects of NaOCl on stem cell survival and differentiation 34 and induces stem cell migration, attachment to the dentin surface, and odontoblastic/osteoblastic differentiation. 44,45 In necrotic cases of immature teeth, it has been verified necrotic pulp tissue colonized by bacterial biofilm and alteration in the apical tissues, such as highly reduced cellularity or lack of cells in the apical papilla and discontinuation or absence of the Hertwig's epithelial root sheath (HERS).…”