Bone, dentin and cementum differentially influence the differentiation of osteoclast-like cells

Abstract Our aim was to investigate how different oral hard tissues determine the differentiation of osteoclast-like cells. Murine macrophage cells were stimulated for 12 day with RANKL and M-CSF on dentin slices. Morphological changes of cells and hard tissues were examined by electron microscopy a...

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Main Authors: Annika Both, Ghosn Ibrahim, Jana Marciniak, Birgit Rath-Deschner, Erika Calvano Küchler, Christian Kirschneck, Lina Gölz, Andreas Jäger, Svenja Beisel-Memmert
Format: Article
Language:English
Published: Nature Portfolio 2025-06-01
Series:Scientific Reports
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Online Access:https://doi.org/10.1038/s41598-025-04874-9
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author Annika Both
Ghosn Ibrahim
Jana Marciniak
Birgit Rath-Deschner
Erika Calvano Küchler
Christian Kirschneck
Lina Gölz
Andreas Jäger
Svenja Beisel-Memmert
author_facet Annika Both
Ghosn Ibrahim
Jana Marciniak
Birgit Rath-Deschner
Erika Calvano Küchler
Christian Kirschneck
Lina Gölz
Andreas Jäger
Svenja Beisel-Memmert
author_sort Annika Both
collection DOAJ
description Abstract Our aim was to investigate how different oral hard tissues determine the differentiation of osteoclast-like cells. Murine macrophage cells were stimulated for 12 day with RANKL and M-CSF on dentin slices. Morphological changes of cells and hard tissues were examined by electron microscopy and toluidine blue staining. Cells were stimulated with RANKL and M-CSF on pulverized bone, dentin, cementum or polystyrene—with and without stimulation. TRAP staining was performed. To elucidate total gene expression, RNA sequencing was carried out. Four target genes (CXCL2, IGF-1, GDF15, HSPA1b) were selected and their expression was analyzed by RT-PCR and ELISA. Statistics comprised One-way ANOVA and Tukey’s test (P < 0.05). Stimulation induced differentiation of mouse macrophages into TRAP-positive osteoclast-like cells forming resorption pits on dentin. Gene expression analysis revealed that 1930, 446 and 87 genes were differentially regulated by cultivation on cementum, bone or dentin respectively compared to polystyrene. Culture on bone or dentin caused CXCL2 upregulation. In all stimulated groups IGF-1 was downregulated while GDF15 expression was elevated in cultures on dentin. Cultivation of cells on cementum resulted in an upregulated HSPA1b expression. Our results indicate that extracellular matrix of different oral hard tissues plays an important role in differentiation processes of osteoclast-like cells.
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spelling doaj-art-37fb9cf30c3442f0a2cbb98172a9cc922025-08-20T03:10:36ZengNature PortfolioScientific Reports2045-23222025-06-0115111110.1038/s41598-025-04874-9Bone, dentin and cementum differentially influence the differentiation of osteoclast-like cellsAnnika Both0Ghosn Ibrahim1Jana Marciniak2Birgit Rath-Deschner3Erika Calvano Küchler4Christian Kirschneck5Lina Gölz6Andreas Jäger7Svenja Beisel-Memmert8Department of Orthodontics, Medical Faculty, University Hospital BonnDepartment of Orthodontics, Medical Faculty, University Hospital BonnDepartment of Orthodontics, Medical Faculty, University Hospital BonnDepartment of Orthodontics, Medical Faculty, University Hospital BonnDepartment of Orthodontics, Medical Faculty, University Hospital BonnDepartment of Orthodontics, Medical Faculty, University Hospital BonnDepartment of Orthodontics and Orofacial Orthopedics, Friedrich-Alexander-University Erlangen-NürnbergDepartment of Orthodontics, Medical Faculty, University Hospital BonnDepartment of Orthodontics, Medical Faculty, University Hospital BonnAbstract Our aim was to investigate how different oral hard tissues determine the differentiation of osteoclast-like cells. Murine macrophage cells were stimulated for 12 day with RANKL and M-CSF on dentin slices. Morphological changes of cells and hard tissues were examined by electron microscopy and toluidine blue staining. Cells were stimulated with RANKL and M-CSF on pulverized bone, dentin, cementum or polystyrene—with and without stimulation. TRAP staining was performed. To elucidate total gene expression, RNA sequencing was carried out. Four target genes (CXCL2, IGF-1, GDF15, HSPA1b) were selected and their expression was analyzed by RT-PCR and ELISA. Statistics comprised One-way ANOVA and Tukey’s test (P < 0.05). Stimulation induced differentiation of mouse macrophages into TRAP-positive osteoclast-like cells forming resorption pits on dentin. Gene expression analysis revealed that 1930, 446 and 87 genes were differentially regulated by cultivation on cementum, bone or dentin respectively compared to polystyrene. Culture on bone or dentin caused CXCL2 upregulation. In all stimulated groups IGF-1 was downregulated while GDF15 expression was elevated in cultures on dentin. Cultivation of cells on cementum resulted in an upregulated HSPA1b expression. Our results indicate that extracellular matrix of different oral hard tissues plays an important role in differentiation processes of osteoclast-like cells.https://doi.org/10.1038/s41598-025-04874-9Orthodontic root resorptionOsteoclast differentiationCXCL2IGF-1GDF15HSPA1b
spellingShingle Annika Both
Ghosn Ibrahim
Jana Marciniak
Birgit Rath-Deschner
Erika Calvano Küchler
Christian Kirschneck
Lina Gölz
Andreas Jäger
Svenja Beisel-Memmert
Bone, dentin and cementum differentially influence the differentiation of osteoclast-like cells
Scientific Reports
Orthodontic root resorption
Osteoclast differentiation
CXCL2
IGF-1
GDF15
HSPA1b
title Bone, dentin and cementum differentially influence the differentiation of osteoclast-like cells
title_full Bone, dentin and cementum differentially influence the differentiation of osteoclast-like cells
title_fullStr Bone, dentin and cementum differentially influence the differentiation of osteoclast-like cells
title_full_unstemmed Bone, dentin and cementum differentially influence the differentiation of osteoclast-like cells
title_short Bone, dentin and cementum differentially influence the differentiation of osteoclast-like cells
title_sort bone dentin and cementum differentially influence the differentiation of osteoclast like cells
topic Orthodontic root resorption
Osteoclast differentiation
CXCL2
IGF-1
GDF15
HSPA1b
url https://doi.org/10.1038/s41598-025-04874-9
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