Objective. To evaluate the push-out bond strength of premixed and powder-liquid bioceramic sealers with or without gutta-percha (GP) cone. Materials and Methods. Radicular dentin samples were prepared from 80 single-rooted human teeth. After root canal preparation using ProTaper® and irrigation with NaOCl and EDTA, teeth were divided according to the root canal sealer ( n = 20 ): AH Plus®, EndoSequence® BC Sealer™, ProRoot® Endo Sealer, and BioRoot™ RCS. Samples were randomly divided into two subgroups ( n = 10 ): GP-S: root canal filling using the single-cone technique, or S: filling with only sealer. Specimens were kept at 37°C and 100% humidity in calcium-free PBS for 30 days. The push-out bond strength was measured in MPa. Fractured specimens were observed at 25x to evaluate the type of failure. pH and calcium ion release were measured at different experimental periods. Raman and SEM-EDAX analyses were performed for root canal sealers. Data were analysed using three-way analysis of variance (ANOVA) and post hoc Tukey test at a significance of P < 0.05 . Results. Push-out bond strength was greater for samples obturated with only sealers (S) than samples obturated with the single-cone technique (GP-S) ( P < 0.05 ). BioRoot™ RCS had greater bond strength than EndoSequence® BC Sealer™. Adhesive failures between cement and gutta-percha cone (87.5%) were predominant in the GP-S. Cohesive failures were predominant for S (80%). BioRoot™ RCS and ProRoot® ES presented higher alkalinization potential than the premixed sealer (EndoSequence® BC Sealer™). Powder-liquid bioceramic sealers (BioRoot™ RCS and ProRoot® ES) released the highest cumulative amount of calcium (28.46 mg/L and 20.05 mg/L). Conclusion. Push-out test without gutta-percha cone presents higher bond strength for bioceramic sealers. Powder-liquid calcium silicate-based sealers present greater bioactivity related to alkalinization potential and calcium ion release.

The biomineralisation of radicular dentin involves complex molecular signalling. Providing evidence of protein binding sites for calcium ions and mineral precipitation is essential for a better understanding of the remineralisation process. This study aimed to evaluate the functional relationship of metalloproteinases (MMPs) and non-collagenous proteins (NCPs) with mineral initiation and maturation during the biomineralisation of radicular dentin. A standardized demineralisation procedure was performed to radicular dentin slices. Samples were remineralised in a PBS-bioactive material system for different periods of time. Assessments of ion exchange, Raman analysis, and energy dispersive X-ray analysis (EDAX) with a scanning electron microscope (SEM) were used to evaluate the remineralisation process. Immunohistochemistry and zymography were performed to analyse NCPs and MMPs expression. SEM evaluation showed that the mineral nucleation and growth occurs, exclusively, on the demineralised radicular dentin surface. Raman analysis of remineralised dentin showed intense peaks at 955 and 1063 cm−1, which can be attributed to carbonate apatite formation. Immunohistochemistry of demineralised samples revealed the presence of DMP1-CT, mainly in intratubular dentin, whereas DSPP in intratubular and intertubular dentin. DMP1-CT and DSPP binding sites control carbonate apatite nucleation and maturation guiding the remineralisation of radicular dentin.

Apical periodontitis is frequently associated with the presence of bacteria biofilm, which has an indisputable impact on the prognosis of endodontic therapy due to the high resistance to adverse environmental conditions, chemicals, and antibiotic therapy that characterize bacteria within biofilm. The biofilm matrix acts as a protective shield over the encased microorganisms. The aim of this investigation was to identify the main biochemical components of biofilm matrix from endodontic mono- and dual-species biofilms. Enterococcus faecalis and Actinomyces naeslundii were cultured as mono- and dual-species biofilms for 14 days. Crude extracellular polymeric substances (EPSs) from biofilm matrices were extracted using chemical and physical methods. High-performance liquid chromatography, gas chromatography, and mass spectrometry were used to determine the carbohydrate, protein, and fatty acid components. Chemical analysis of the biofilm matrices revealed that they were mainly composed of stachyose, maltose, and mannose carbohydrates. The protein profile in all biofilm samples showed abundant oxidoreductases and chaperone proteins and some virulence- associated proteins mainly located in the membrane surface. High percentages of saturated and monounsaturated fatty acids were identified in all biofilm matrices, with a major prevalence of palmitic, stearic, and oleic acids. Based on the results, it was possible to obtain for the first time a general overview of the biochemical profile of endodontic biofilm matrices.

Highlights of the Study• Radicular dentin matrix metalloproteinases have potential implications on regenerative therapies.• Endodontic irrigant solutions affect the expression of matrix metalloproteinases on radicular dentin.• Sodium hypochlorite and chlorhexidine downregulate the expression of matrix metalloproteinases.• Conditioning with ethylenediaminetetraacetic acid could improve the environment for dentin remineralization.

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