Hydraulic calcium silicate-based root canal sealers mitigate proinflammatory cytokine synthesis and promote osteogenesis in vitro

Aseel Alchawoosh; Kentaro Hashimoto; Nobuyuki Kawashima; Sonoko Noda; Kosuke Nozaki; Takashi Okiji

doi:10.1016/j.jds.2022.12.019

Hydraulic calcium silicate-based root canal sealers mitigate proinflammatory cytokine synthesis and promote osteogenesis in vitro

J Dent Sci. 2023 Oct;18(4):1731-1739. doi: 10.1016/j.jds.2022.12.019. Epub 2023 Jan 7.

Authors

Aseel Alchawoosh¹, Kentaro Hashimoto¹, Nobuyuki Kawashima¹, Sonoko Noda¹, Kosuke Nozaki², Takashi Okiji¹

Affiliations

¹ Department of Pulp Biology and Endodontics, Division of Oral Health Sciences, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, Tokyo, Japan.
² Department of Advanced Prosthodontics, Division of Oral Health Sciences, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, Tokyo, Japan.

Abstract

Background/purpose: The mineralized tissue-inductive ability and anti-inflammatory properties of hydraulic calcium silicate-based (HCSB) sealers have not been fully elucidated. This study aimed to evaluate the effects of the HCSB sealers Bio-C sealer (BioC), Well-Root ST (WST), and EndoSequence BC sealer (BC), on osteoblastic differentiation/mineralization and proinflammatory cytokine synthesis by macrophages.

Materials and methods: Diluted extracts of set sealers or calcium chloride solutions of approximately equivalent Ca²⁺ concentrations were applied to a mouse osteoblastic cell line (Kusa-A1 cells) and lipopolysaccharide-stimulated mouse macrophage cell line (RAW264.7 cells). Expressions of osteoblastic markers in Kusa-A1 cells and proinflammatory cytokines in RAW264.7 cells were evaluated by reverse transcription-quantitative polymerase chain reaction and enzyme-linked immunosorbent assays. Mineralized nodules were detected by Alizarin red S staining. Cell proliferation was assessed by WST-8 assay and cell attachment on set sealers was examined by scanning electron microscopy.

Results: The three sealer extracts significantly upregulated osteocalcin and osteopontin mRNA, and promoted significant mineralized nodule formation in Kusa-A1 cells. The three sealer extracts significantly downregulated the mRNA expressions of interleukin (IL)-1α, IL-1β, IL-6, and tumor necrosis factor (TNF)-α and protein levels of IL-6 and TNF-α in RAW264.7 cells. Calcium chloride solutions induced osteoblastic differentiation/mineralization. AH Plus Jet (a control sealer) extract did not. The three HCSB sealers did not interfere with the growth and attachment of Kusa-A1 cells.

Conclusion: BioC, WST, and BC were biocompatible, upregulated osteoblastic differentiation/mineralization, and downregulated proinflammatory cytokine expression. Ca²⁺ released from HCSB sealers might be involved, at least in part, in the induction of osteoblastic differentiation/mineralization.

Keywords: Calcium signaling; Cytokine; Differentiation; Endodontic obturation; Macrophage; Osteoblast.