A Novel Laboratorial Approach to Evaluate Bacterial Microleakage of Endodontic Sealers
Emannuel Fonseca Thomé da Silva Monteiro, Alice Abib Ramos Fabri, Marcelo Barbosa Garcia, Sabrina Silva Azevedo, Higor Machado Gonçalves, Ana Clara Miranda Porto Figueiredo, Fabrícia Nogueira Klein, Gabriela Ceccon Chianca, Roberta Barcelos, Helvécio Cardoso Corrêa Póvoa, Leonardo Santos Antunes, Natalia Lopes Pontes Póvoa IorioAbstract
To avoid failure of endodontic treatment, it is important to properly seal the root canals after chemomechanical preparation in order to avoid microleakage. Typically, sealing involves the use of gutta‐percha and sealing cement, the latter of which adheres to the dentin and fills the spaces between the gutta‐percha and the dentin walls. Laboratory studies have been performed to detect microleakage of endodontic sealers based on various criteria, including microbiological ones; however, none guaranteed that the leakage occurred through the root canal space and not through other pathways. This article proposes a new laboratory assay to evaluate bacterial microleakage in endodontic sealers, ensuring visualization of the correct inoculum infiltration pathway. Furthermore, we present the main critical parameters and provide suggestions on how to manage them. These protocols allow the detection of bacterial microleakage in bovine teeth filled with endodontic sealer and inoculated with Enterococcus faecalis . © 2026 The Author(s). Current Protocols published by Wiley Periodicals LLC.
Basic Protocol 1 : Bovine teeth selection and cleaning
Basic Protocol 2 : Bovine teeth inclusion
Basic Protocol 3 : Tooth standardization
Basic Protocol 4 : Set preparation
Basic Protocol 5 : Inoculum preparation
Basic Protocol 6 : Canal inoculation
Basic Protocol 7 : Set incubation