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. 2025 Feb 18;29(2):136.
doi: 10.1007/s00784-025-06231-6.

Impact of endodontic irrigant solutions on the viability and metabolism of stem cells from apical papilla: proposal of a novel in vitro model

Letycia Accioly Simões Coelho  1   2 , Ester Alves Ferreira Bordini  3 , Fernanda Balestrero Cassiano  3 , Vitor de Toledo Stuani  3 , Renato Menezes Silva  4 , Diana Gabriela Soares Dos Passos  3 , Marco Antonio Hungaro Duarteg  3
Affiliations

Impact of endodontic irrigant solutions on the viability and metabolism of stem cells from apical papilla: proposal of a novel in vitro model

Letycia Accioly Simões Coelho et al. Clin Oral Investig. .

Abstract

Objectives: This study aimed to evaluate the effect of different irrigation protocols on the viability and metabolism of stem cells from the apical papilla (SCAP) using a new in vitro protocol that simulates the clinical situation.

Materials and methods: Forty-eight bovine dentin cylinders were obtained and prepared to simulate teeth with incomplete rhizogenesis, positioned under a three-dimensional (3D) culture of SCAPs to mimic the apical papilla. The cylinders were divided into four groups (n = 8) according to the irrigating solution: Control; NaOCl (Sodium hypochlorite 1%); EDTA (17% ethylenediaminetetraacetic acid); and NaOCl + EDTA. Subsequently, the viability (Live/Dead n = 2) and metabolism (Alamar Blue n = 6) of the cells were assessed (ISO 10993). Data were analyzed using Kruskal-Wallis and Dunn tests (p < 0.05).

Results: In the 1 to 3 days period, Control and EDTA had significantly higher increases in metabolism compared to NaOCl and NaOCl + EDTA (p < 0.05). In the 3- to 7-day period, metabolism significantly decreased in NaOCl + EDTA compared to EDTA (p < 0.05) but was similar to Control and NaOCl. Additionally, significant differences were observed within groups Control, EDTA, and NaOCl + EDTA across the two periods (p < 0.05).

Conclusions: The tested in vitro model allows for the analysis of the response of SCAPs to different irrigating solutions, simulating the clinical situation. Sodium hypochlorite 1% demonstrated high cytotoxicity to SCAPs, whose effects were partially reversed by 17% EDTA.

Clinical relevane: The methodology developed provides a tool for future investigations, allowing for the assessment of new irrigants and techniques that may optimize tissue regeneration.

Keywords: EDTA; Regenerative endodontics; Sodium hypochlorite; Stem cells rom apical papilla.

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Conflict of interest statement

Declarations. Ethical approval: This study was approved by the Animal Research Ethics Committee and the Human Research Ethics Committee and was conducted following the the guidelines of the International Organization for Standardization (ISO 10993) [20]. Informed consent: Not applicable. Competing interests: The authors declare no competing interests.

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