Animal testing is crucial in situations when research on humans is not allowed because of unknown health risks and ethical concerns. The current project aims to develop reporting guidelines exclusively for animal studies in Endodontology, using an established consensus‐based methodology. The guidelines have been named: Preferred Reporting Items for Animal Studies in Endodontology (PRIASE) 2021. Nine individuals (PD, VN, AK, PM, MN, JF, EP, JJ, SJ), including the project leaders (PD, VN) formed a steering committee. The steering committee developed a novel checklist by adapting and integrating their animal testing and peer‐review experience with the Animals in Research: Reporting In Vivo Experiments (ARRIVE) guidelines and also the Clinical and Laboratory Images in Publications (CLIP) principles. A PRIASE Delphi Group (PDG) and PRIASE Online Meeting Group (POMG) were also formed. Thirty‐one PDG members participated in the online Delphi process and achieved consensus on the checklist items and flowchart that were used to formulate the PRIASE guidelines. The novel PRIASE 2021 guidelines were discussed with the POMG on 9^th September 2020 via a Zoom online video call attended by 21 individuals from across the globe, and seven steering committee members. Following the discussions, the guidelines were modified and then piloted by several authors whilst writing a manuscript involving research on animals. The PRIASE 2021 guidelines are a checklist consisting of 11 domains and 43 individual items together with a flowchart. The PRIASE 2021 guidelines are focused on improving the methodological principles, reproducibility and quality of animal studies in order to enhance their reliability as well as repeatability to estimate the effects of endodontic treatments and usefulness for guiding future clinical studies on humans.

Laws and ethics require that before conducting human clinical trials, a new material, device or drug may have to undergo testing in animals in order to minimize health risks to humans, unless suitable supporting grandfather data already exist. The Preferred Reporting Items for Animal Studies in Endodontology (PRIASE) 2021 guidelines were developed exclusively for the specialty of Endodontology by integrating and adapting the ARRIVE (Animals in Research: Reporting In Vivo Experiments) guidelines and the Clinical and Laboratory Images in Publications (CLIP) principles using a validated consensus‐based methodology. Implementation of the PRIASE 2021 guidelines will reduce potential sources of bias and thus improve the quality, accuracy, reproducibility, completeness and transparency of reports describing animal studies in Endodontology. The PRIASE 2021 guidelines consist of a checklist with 11 domains and 43 individual items and a flowchart. The aim of the current document is to provide an explanation for each item in the PRIASE 2021 checklist and flowchart and is supplemented with examples from the literature in order for readers to understand their significance and to provide usage guidance. A link to the PRIASE 2021 explanation and elaboration document and PRIASE 2021 checklist and flowchart is available on the Preferred Reporting Items for study Designs in Endodontology (PRIDE) website (http://pride‐endodonticguidelines.org/priase/).

Background and objectives: Pulp stones are hard tissue structures formed in the pulp of permanent and deciduous teeth. Few studies have evaluated their morphology and chemical composition. However, their formation, composition, configuration and role played in overall health status are still unclear. Clinically, they may be symptomatic; technically, they impede access during endodontic therapy, increasing the risk of treatment errors. Thus, this study aimed to morphologically analyze pulp stones and present their chemical quantification, identifying their main chemical elements. It also correlates the results with their possible induction mechanisms. Materials and Methods: Seven pulp nodules were collected from molar teeth needing endodontic treatment. The morphology of the stones was analyzed by scanning electron microscopy (SEM), and their chemical composition was determined by X-ray dispersive energy spectroscopy (EDX). Results: These structures varied considerably in shape, size and topography. The site of the stones in the pulp cavity was the factor that most affected the morphology. The majority of the stones found in the pulp chambers presented nodular morphology, while those in the root canals presented a diffuse shape, resembling root canal anatomy. The topography of the nodules showed heterogeneous relief, revealing smooth and compact areas contrasting with the rugged and porous ones. The chemical composition varied depending on the location of the nodule in the pulp cavity and the relief of the analyzed area. Radicular stones presented considerably lower calcium and phosphorus content than coronary nodules. Conclusions: The high cellularity rate of the coronal pulp predisposes this region to nodular mineralizations around injured cells. The presence of larger caliber vascular bundles and higher collagen fiber content in radicular pulp determines a diffuse morphological pattern in this region. Understanding the morphology and chemical composition of the pulp stones allows future translational pathways towards the prevention or treatment of such conditions.

Bacteriophages, viruses capable of killing bacteria, were discovered in 1915, but the interest in their study has been limited since the advent of antibiotics. Their use in dentistry is still very limited. The authors reviewed studies about bacteriophage structure, mode of action, uses in oral health, and possible future uses in dentistry associated with their possible action over biofilm, as well as the advantages and limitations of phage therapy.

Biofilms are composed of multicellular communities of microbial cells and their self-secreted extracellular polymeric substances (EPS). The viruses named bacteriophages can infect and lyze bacterial cells, leading to efficient biofilm eradication. The aim of this study was to analyze how bacteriophages disrupt the biofilm structure by killing bacterial cells and/or by damaging extracellular polysaccharides, proteins, and DNA. The use of colorimetric and spectrofluorimetric methods and confocal laser scanning microscopy (CLSM) enabled a comprehensive assessment of phage activity against E. faecalis biofilms. The impact of the phages vB_Efa29212_2e and vB_Efa29212_3e was investigated. They were applied separately or in combination on 1-day and 7-day-old biofilms. Phages 2e effectively inhibited the growth of planktonic cells with a limited effect on the biofilm. They did not notably affect extracellular polysaccharides and proteins; however, they increased DNA levels. Phages 3e demonstrated a potent and dispersing impact on E. faecalis biofilms, despite being slightly less effective than bacteriophages 2e against planktonic cells. Phages 3e reduced the amount of extracellular polysaccharides and increased eDNA levels in both 1-day-old and 7-day-old biofilm cultures. Phage cocktails had a strong antimicrobial effect on both planktonic and biofilm-associated bacteria. A significant reduction in the levels of polysaccharides, proteins, and eDNA in 1-day-old biofilm samples was noted, which confirms that phages interfere with the structure of E. faecalis biofilm by killing bacterial cells and affecting extracellular polymer levels.