Osteogenic Potential of Various Premixed Hydraulic Calcium Silicate-Based Sealers on Human Bone Marrow Stem Cells

This study aims to compare the osteogenic potential of premixed hydraulic calcium silicate-based sealers (HCSSs) with an epoxy resin-based sealer in human bone marrow-derived stem cells (hBMSCs). Three HCSSs (White Endoseal MTA, One-Fil, EndoSequence BC Sealer) were compared with AH Plus Jet, an epoxy resin-based sealer. Disk-shaped specimens were prepared using sterilized Teflon tubes and immersed in osteogenic medium to create eluates. hBMSCs were cultured in each eluate, and osteogenic potential was assessed by alkaline phosphatase (ALP) activity (n = 6), Alizarin Red-S (ARS) staining (n = 6), quantitative real-time polymerase chain reaction (qPCR) (n = 3), and Western blot analysis. Statistical analyses were conducted using SPSS (version 24.0), with significance set at <i>p</i> < 0.05. All experimental groups exhibited higher ALP activity than the control on day 4. ARS staining of HCSSs differed significantly from AH Plus Jet on day 14 (<i>p</i> < 0.05), while White Endoseal MTA exhibited the highest intensity. qPCR revealed that EndoSequence BC Sealer induced the highest <i>SMAD1</i> expression on day 4, while One-Fil and EndoSequence BC Sealer significantly upregulated <i>RUNX2</i> expression compared with AH Plus Jet (<i>p</i> < 0.05). Western blotting confirmed that EndoSequence BC Sealer induced the highest RUNX2 protein expression. Collectively, premixed HCSSs promoted superior mineralization and <i>RUNX2</i> expression compared to conventional resin-based sealer in hBMSCs.