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Mineral trioxide aggregate immersed in sodium hypochlorite reduce the osteoblastic differentiation of human periodontal ligament stem cells.

ABSTRACT: White mineral trioxide aggregate (WMTA) is a root canal treatment material, which is known to exhibit a dark brown color when in contact with sodium hypochlorite solution (NaOCl). This study aimed to investigate the effects of NaOCl on the surface properties of WMTA discs and WMTA-induced osteoblastic differentiation of periodontal ligament stem cells (PDLSCs). Mixed WMTA (ProRoot MTA) was filled into the molds to form WMTA discs. These discs were immersed in distilled water (D-WMTA) or 5% NaOCl (Na-WMTA). Their surface structures and Ca²⁺ release level was investigated. Moreover, they were cultured with a clonal human PDLSC line (line 1-17 cells). The main crystal structures of Na-WMTA were identical to the structures of D-WMTA. Globular aggregates with polygonal and needle-like crystals were found on D-WMTA and Na-WMTA, which included Ca, Si, Al, C and O. However, many amorphous structures were also identified on Na-WMTA. These structures consisted of Na and Cl, but did not include Ca. NaOCl immersion also reduced Ca²⁺ release level from whole WMTA discs. Line 1-17 cells cultured with D-WMTA formed many mineralized nodules and exhibited high expression levels of osteoblast-related genes. However, cells incubated with Na-WMTA generated a small number of nodules and showed low expression levels of osteoblast-related genes. These results indicated that NaOCl reduced Ca²⁺ release from WMTA by generating amorphous structures and changing its elemental distribution. NaOCl may also partially abolish the ability of WMTA to stimulate osteoblastic differentiation of PDLSCs.

SUBMITTER: Yamashita K

PROVIDER: S-EPMC8585953 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:Background and objectiveMineral trioxide aggregate (MTA) is a biomaterial used in endodontic procedures as it exerts beneficial effects on regenerative processes. In this study, we evaluate the effect of MTA on healing of periodontal ligament (PDL) and surrounding tissue, following injury, in a transgenic mouse model and on the differentiation of murine mesenchymal progenitor cells in vitro.Material and methodsWe used an inducible Cre-loxP in vivo fate mapping approach to examine the effects of MTA on the contributions of descendants of cells expressing the αSMA-CreERT2 transgene (SMA9+ ) to the PDL and alveolar bone after experimental injury to the root furcation on the maxillary first molars. Col2.3GFP was used as a marker to identify mature osteoblasts, cementoblasts and PDL fibroblasts. The effects of MTA were examined 2, 17 and 30 days after injury and compared histologically with sealing using an adhesive system. The effects of two dilutions of medium conditioned with MTA on proliferation and differentiation of mesenchymal progenitor cells derived from bone marrow (BMSC) and periodontal ligament (PDLC) in vitro were examined using the PrestoBlue viability assay, alkaline phosphatase and Von Kossa staining. The expression of markers of differentiation was assessed using real-time PCR.ResultsHistological analyses showed better repair in teeth restored with MTA, as shown by greater expansion of SMA9+ progenitor cells and Col2.3GFP+ osteoblasts compared with control teeth. We also observed a positive effect on differentiation of SMA9+ progenitors into osteoblasts and cementoblasts in the apical region distant from the site of injury. The in vitro data showed that MTA-conditioned medium reduced cell viability and osteogenic differentiation in both PDLC and BMSC, indicated by reduced von Kossa staining and lower expression of osteocalcin and bone sialoprotein. In addition, cultures grown in the presence of MTA had marked decreases in SMA9+ and Col2.3GFP+ areas as compared with osteogenic medium, confirming reduced osteogenesis.ConclusionMTA promotes regeneration of injured PDL and alveolar bone, reflected as contribution of progenitors (SMA9+ cells) into osteoblasts (Col2.3GFP+ cells). In vitro, MTA-conditioned medium fails to promote osteogenic differentiation of both PDLC and BMSC.

Project description:BackgroundMineral trioxide aggregate (MTA) is a commonly used endodontic biomaterial. The physicochemical properties of MTA have a crucial role in designating clinical outcome, and different factors can affect these properties. Various methods have been used for mixing MTA, including manual, mechanical, and ultrasonic. The aim of this systematic review was to evaluate the effect of different mixing methods on the physicochemical properties of MTA.Materials and methodsElectronic databases including PubMed, Embase, Web of Science, and Scopus were searched up to May 2022. In order to cover gray literature, the ProQuest and Google Scholar databases were also searched to detect theses and conference proceedings. For quality assessment of the included studies, we used a modified version of the Cochrane risk of bias tool for randomized controlled trials (RCTs). Experimental studies which had assessed at least one property of MTA and compared at least two different mixing methods of MTA were included in this study. All animal studies, reviews, case reports, and case series were excluded.ResultsFourteen studies were included. The results showed that the ultrasonic mixing method significantly improved some MTA characteristics, including microhardness, flowability, solubility, setting time, and porosity. However, the mechanical mixing method improved other properties including flowability, solubility, push-out bond strength, and hydration. The manual mixing method showed inferior results compared to other mixing methods in terms of microhardness, flowability, solubility, setting time, push-out bond strength, porosity, and hydration. Different mixing methods had a similar effect on compressive strength, sealing ability, pH and calcium ion release, volume change, film thickness, and flexural strength of MTA.ConclusionMechanical and ultrasonic mixing methods are superior to the manual mixing method in terms of improving physicochemical properties of MTA. No report of selection bias and varieties in methodologies were limitations of evidence.

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Mineral trioxide aggregate immersed in sodium hypochlorite reduce the osteoblastic differentiation of human periodontal ligament stem cells.

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