Ruxolitinib altered IFN-β induced necroptosis of human dental pulp stem cells during osteoblast differentiation

Atsuko Tanaka; Satoru Hayano; Masayo Nagata; Takahiro Kosami; Ziyi Wang; Hiroshi Kamioka

doi:10.1016/j.archoralbio.2023.105797

Ruxolitinib altered IFN-β induced necroptosis of human dental pulp stem cells during osteoblast differentiation

Arch Oral Biol. 2023 Nov:155:105797. doi: 10.1016/j.archoralbio.2023.105797. Epub 2023 Aug 22.

Authors

Atsuko Tanaka¹, Satoru Hayano², Masayo Nagata³, Takahiro Kosami³, Ziyi Wang⁴, Hiroshi Kamioka³

Affiliations

¹ Department of Orthodontics, Okayama University Hospital, Okayama, Japan.
² Department of Orthodontics, Okayama University Hospital, Okayama, Japan. Electronic address: shayano@okayama-u.ac.jp.
³ Department of Orthodontics, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University, Okayama, Japan.
⁴ Department of Molecular Biology and Biochemistry, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama, Japan; Research Fellow of Japan Society for the Promotion of Science, Tokyo, Japan.

PMID: 37633030
DOI: 10.1016/j.archoralbio.2023.105797

Abstract

Objective: This study aimed to evaluate the role of ruxolitinib in the interferon beta (IFN-β) mediated osteoblast differentiation using human dental pulp stem cells (hDPSCs).

Design: hDPSCs from five deciduous teeth of healthy patients were stimulated by adding human recombinant IFN-β protein (1 or 2 ng/ml) to the osteogenic differentiation induction medium. Substrate formation was determined using Alizarin Red staining, calcium concentration, and osteoblast marker expression levels. Ruxolitinib was used to inhibit the Janus kinase/signal transducers and activators of transcription (JAK-STAT) pathway. Apoptosis was detected using terminal deoxynucleotidyl nick-end labeling (TUNEL) staining, and necroptosis was detected using propidium iodide staining and phosphorylated mixed lineage kinase domain-like protein (pMLKL) expression.

Results: In the IFN-β-treated group, substrate formation was inhibited by a reduction in alkaline phosphatase (ALP) expression in a concentration-dependent manner. Although the proliferation potency was unchanged between the IFN-β-treated and control groups, the cell number was significantly reduced in the experimental group. TUNEL-positive cell number was not significantly different; however, the protein level of necroptosis markers, interleukin-6 (IL-6) and pMLKL were significantly increased in the substrate formation. Cell number and ALP expression level were improved in the group administered ruxolitinib, a JAK-STAT inhibitor. Additionally, ruxolitinib significantly suppressed IL-6 and pMLKL levels.

Conclusion: Ruxolitinib interfered with the IFN-β-mediated necroptosis and osteogenic differentiation via the JAK-STAT pathway.

Keywords: Janus kinase/signal transducers and activators of transcription pathway; Necroptosis; Osteoblast; Singleton-Merten Syndrome; Type-I interferon.