Extracellular Vesicles Derived From Murine Cementoblasts Possess the Potential to Increase Receptor Activator of Nuclear Factor-κB Ligand-Induced Osteoclastogenesis

Cementum resorption, unlike bone resorption, is clinically known to occur only with limited pathological stimuli, such as trauma, orthodontic forces, and large apical periodontitis; however, the molecular mechanisms that control osteoclast formation on the cementum surface remain unclear. In this st...

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Main Authors: Rei Sato, Kentaro Maruyama, Eiji Nemoto, Yukihiko Sakisaka, Shigeki Suzuki, Jiajun Li, Kento Numazaki, Hiroyuki Tada, Satoru Yamada
Format: Article
Language:English
Published: Frontiers Media S.A. 2022-02-01
Series:Frontiers in Physiology
Subjects:
Online Access:https://www.frontiersin.org/articles/10.3389/fphys.2022.825596/full
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author Rei Sato
Kentaro Maruyama
Eiji Nemoto
Yukihiko Sakisaka
Shigeki Suzuki
Jiajun Li
Kento Numazaki
Hiroyuki Tada
Satoru Yamada
author_facet Rei Sato
Kentaro Maruyama
Eiji Nemoto
Yukihiko Sakisaka
Shigeki Suzuki
Jiajun Li
Kento Numazaki
Hiroyuki Tada
Satoru Yamada
author_sort Rei Sato
collection DOAJ
description Cementum resorption, unlike bone resorption, is clinically known to occur only with limited pathological stimuli, such as trauma, orthodontic forces, and large apical periodontitis; however, the molecular mechanisms that control osteoclast formation on the cementum surface remain unclear. In this study, we focused on extracellular vesicles (EVs) secreted by cementoblasts and analyzed their effects on osteoclast differentiation. EVs were extracted from the conditioned medium (CM) of the mouse cementoblast cell line OCCM-30. Transmission electron microscopy (TEM) analysis confirmed the presence of EVs with a diameter of approximately 50–200 nm. The effect of the EVs on osteoclast differentiation was examined using the mouse osteoclast progenitor cell line RAW 264.7 with recombinant receptor activator of nuclear factor (NF)-κB ligand (rRANKL) stimulation. EVs enhanced the formation of tartrate-resistant acid phosphatase (TRAP) activity-positive cells upon rRANKL stimulation. EVs also enhanced the induction of osteoclast-associated gene and protein expression in this condition, as determined by real-time PCR and Western blotting, respectively. On the other hand, no enhancing effect of EVs was observed without rRANKL stimulation. A Western blot analysis revealed no expression of receptor activator of NF-κB ligand (RANKL) in EVs themselves. The effect on rRANKL-induced osteoclast differentiation was examined using the CM of cementoblasts in terms of TRAP activity-positive cell formation and osteoclast-associated gene expression. The conditioned medium partly inhibited rRANKL-induced osteoclast differentiation and almost completely suppressed its enhancing effect by EVs. These results indicate that cementoblasts secreted EVs, which enhanced RANKL-induced osteoclast differentiation, and simultaneously produced soluble factors that neutralized this enhancing effect of EVs, implicating this balance in the regulation of cementum absorption. A more detailed understanding of this crosstalk between cementoblasts and osteoclasts will contribute to the development of new therapies for pathological root resorption.
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spelling doaj.art-be48275368b241b99a60e74a1d97fdd72022-12-22T00:04:59ZengFrontiers Media S.A.Frontiers in Physiology1664-042X2022-02-011310.3389/fphys.2022.825596825596Extracellular Vesicles Derived From Murine Cementoblasts Possess the Potential to Increase Receptor Activator of Nuclear Factor-κB Ligand-Induced OsteoclastogenesisRei Sato0Kentaro Maruyama1Eiji Nemoto2Yukihiko Sakisaka3Shigeki Suzuki4Jiajun Li5Kento Numazaki6Hiroyuki Tada7Satoru Yamada8Division of Periodontology and Endodontology, Tohoku University Graduate School of Dentistry, Sendai, JapanDivision of Periodontology and Endodontology, Tohoku University Graduate School of Dentistry, Sendai, JapanDivision of Periodontology and Endodontology, Tohoku University Graduate School of Dentistry, Sendai, JapanDivision of Periodontology and Endodontology, Tohoku University Graduate School of Dentistry, Sendai, JapanDivision of Periodontology and Endodontology, Tohoku University Graduate School of Dentistry, Sendai, JapanDivision of Periodontology and Endodontology, Tohoku University Graduate School of Dentistry, Sendai, JapanDivision of Orthodontics and Dentofacial Orthopedics, Tohoku University Graduate School of Dentistry, Sendai, JapanDivision of Oral Immunology, Tohoku University Graduate School of Dentistry, Sendai, JapanDivision of Periodontology and Endodontology, Tohoku University Graduate School of Dentistry, Sendai, JapanCementum resorption, unlike bone resorption, is clinically known to occur only with limited pathological stimuli, such as trauma, orthodontic forces, and large apical periodontitis; however, the molecular mechanisms that control osteoclast formation on the cementum surface remain unclear. In this study, we focused on extracellular vesicles (EVs) secreted by cementoblasts and analyzed their effects on osteoclast differentiation. EVs were extracted from the conditioned medium (CM) of the mouse cementoblast cell line OCCM-30. Transmission electron microscopy (TEM) analysis confirmed the presence of EVs with a diameter of approximately 50–200 nm. The effect of the EVs on osteoclast differentiation was examined using the mouse osteoclast progenitor cell line RAW 264.7 with recombinant receptor activator of nuclear factor (NF)-κB ligand (rRANKL) stimulation. EVs enhanced the formation of tartrate-resistant acid phosphatase (TRAP) activity-positive cells upon rRANKL stimulation. EVs also enhanced the induction of osteoclast-associated gene and protein expression in this condition, as determined by real-time PCR and Western blotting, respectively. On the other hand, no enhancing effect of EVs was observed without rRANKL stimulation. A Western blot analysis revealed no expression of receptor activator of NF-κB ligand (RANKL) in EVs themselves. The effect on rRANKL-induced osteoclast differentiation was examined using the CM of cementoblasts in terms of TRAP activity-positive cell formation and osteoclast-associated gene expression. The conditioned medium partly inhibited rRANKL-induced osteoclast differentiation and almost completely suppressed its enhancing effect by EVs. These results indicate that cementoblasts secreted EVs, which enhanced RANKL-induced osteoclast differentiation, and simultaneously produced soluble factors that neutralized this enhancing effect of EVs, implicating this balance in the regulation of cementum absorption. A more detailed understanding of this crosstalk between cementoblasts and osteoclasts will contribute to the development of new therapies for pathological root resorption.https://www.frontiersin.org/articles/10.3389/fphys.2022.825596/fullextracellular vesiclescementoblastsRANKLosteoclastogenesiscementum resorption
spellingShingle Rei Sato
Kentaro Maruyama
Eiji Nemoto
Yukihiko Sakisaka
Shigeki Suzuki
Jiajun Li
Kento Numazaki
Hiroyuki Tada
Satoru Yamada
Extracellular Vesicles Derived From Murine Cementoblasts Possess the Potential to Increase Receptor Activator of Nuclear Factor-κB Ligand-Induced Osteoclastogenesis
Frontiers in Physiology
extracellular vesicles
cementoblasts
RANKL
osteoclastogenesis
cementum resorption
title Extracellular Vesicles Derived From Murine Cementoblasts Possess the Potential to Increase Receptor Activator of Nuclear Factor-κB Ligand-Induced Osteoclastogenesis
title_full Extracellular Vesicles Derived From Murine Cementoblasts Possess the Potential to Increase Receptor Activator of Nuclear Factor-κB Ligand-Induced Osteoclastogenesis
title_fullStr Extracellular Vesicles Derived From Murine Cementoblasts Possess the Potential to Increase Receptor Activator of Nuclear Factor-κB Ligand-Induced Osteoclastogenesis
title_full_unstemmed Extracellular Vesicles Derived From Murine Cementoblasts Possess the Potential to Increase Receptor Activator of Nuclear Factor-κB Ligand-Induced Osteoclastogenesis
title_short Extracellular Vesicles Derived From Murine Cementoblasts Possess the Potential to Increase Receptor Activator of Nuclear Factor-κB Ligand-Induced Osteoclastogenesis
title_sort extracellular vesicles derived from murine cementoblasts possess the potential to increase receptor activator of nuclear factor κb ligand induced osteoclastogenesis
topic extracellular vesicles
cementoblasts
RANKL
osteoclastogenesis
cementum resorption
url https://www.frontiersin.org/articles/10.3389/fphys.2022.825596/full
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