Uniaxially fixed mechanical boundary condition elicits cellular alignment in collagen matrix with induction of osteogenesis

Abstract Osteocytes differentiated from osteoblasts play significant roles as mechanosensors in modulating the bone remodeling process. While the well-aligned osteocyte network along the trabeculae with slender cell processes perpendicular to the trabeculae surface is known to facilitate the sensing...

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Main Authors: Jeonghyun Kim, Keiichi Ishikawa, Junko Sunaga, Taiji Adachi
Format: Article
Language:English
Published: Nature Publishing Group 2021-04-01
Series:Scientific Reports
Online Access:https://doi.org/10.1038/s41598-021-88505-z
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spelling doaj-7d8c2b39cd454626b0f95c11b79de3de2021-05-02T11:36:05ZengNature Publishing GroupScientific Reports2045-23222021-04-011111910.1038/s41598-021-88505-zUniaxially fixed mechanical boundary condition elicits cellular alignment in collagen matrix with induction of osteogenesisJeonghyun Kim0Keiichi Ishikawa1Junko Sunaga2Taiji Adachi3Institute for Frontier Life and Medical Sciences, Kyoto UniversityDepartment of Micro Engineering, Graduate School of Engineering, Kyoto UniversityInstitute for Frontier Life and Medical Sciences, Kyoto UniversityInstitute for Frontier Life and Medical Sciences, Kyoto UniversityAbstract Osteocytes differentiated from osteoblasts play significant roles as mechanosensors in modulating the bone remodeling process. While the well-aligned osteocyte network along the trabeculae with slender cell processes perpendicular to the trabeculae surface is known to facilitate the sensing of mechanical stimuli by cells and the intracellular communication in the bone matrix, the mechanisms underlying osteocyte network formation remains unclear. Here, we developed a novel in vitro collagen matrix system exerting a uniaxially-fixed mechanical boundary condition on which mouse osteoblast-like MC3T3-E1 cells were subcultured, evoking cellular alignment along the uniaxial boundary condition. Using a myosin II inhibitor, blebbistatin, we showed that the intracellular tension via contraction of actin fibers contributed to the cellular alignment under the influence of isometric matrix condition along the uniaxially-fixed mechanical boundary condition. Furthermore, the cells actively migrated inside the collagen matrix and promoted the expression of osteoblast and osteocyte genes with their orientations aligned along the uniaxially-fixed boundary condition. Collectively, our results suggest that the intracellular tension of osteoblasts under a uniaxially-fixed mechanical boundary condition is one of the factors that determines the osteocyte alignment inside the bone matrix.https://doi.org/10.1038/s41598-021-88505-z
collection DOAJ
language English
format Article
sources DOAJ
author Jeonghyun Kim
Keiichi Ishikawa
Junko Sunaga
Taiji Adachi
spellingShingle Jeonghyun Kim
Keiichi Ishikawa
Junko Sunaga
Taiji Adachi
Uniaxially fixed mechanical boundary condition elicits cellular alignment in collagen matrix with induction of osteogenesis
Scientific Reports
author_facet Jeonghyun Kim
Keiichi Ishikawa
Junko Sunaga
Taiji Adachi
author_sort Jeonghyun Kim
title Uniaxially fixed mechanical boundary condition elicits cellular alignment in collagen matrix with induction of osteogenesis
title_short Uniaxially fixed mechanical boundary condition elicits cellular alignment in collagen matrix with induction of osteogenesis
title_full Uniaxially fixed mechanical boundary condition elicits cellular alignment in collagen matrix with induction of osteogenesis
title_fullStr Uniaxially fixed mechanical boundary condition elicits cellular alignment in collagen matrix with induction of osteogenesis
title_full_unstemmed Uniaxially fixed mechanical boundary condition elicits cellular alignment in collagen matrix with induction of osteogenesis
title_sort uniaxially fixed mechanical boundary condition elicits cellular alignment in collagen matrix with induction of osteogenesis
publisher Nature Publishing Group
series Scientific Reports
issn 2045-2322
publishDate 2021-04-01
description Abstract Osteocytes differentiated from osteoblasts play significant roles as mechanosensors in modulating the bone remodeling process. While the well-aligned osteocyte network along the trabeculae with slender cell processes perpendicular to the trabeculae surface is known to facilitate the sensing of mechanical stimuli by cells and the intracellular communication in the bone matrix, the mechanisms underlying osteocyte network formation remains unclear. Here, we developed a novel in vitro collagen matrix system exerting a uniaxially-fixed mechanical boundary condition on which mouse osteoblast-like MC3T3-E1 cells were subcultured, evoking cellular alignment along the uniaxial boundary condition. Using a myosin II inhibitor, blebbistatin, we showed that the intracellular tension via contraction of actin fibers contributed to the cellular alignment under the influence of isometric matrix condition along the uniaxially-fixed mechanical boundary condition. Furthermore, the cells actively migrated inside the collagen matrix and promoted the expression of osteoblast and osteocyte genes with their orientations aligned along the uniaxially-fixed boundary condition. Collectively, our results suggest that the intracellular tension of osteoblasts under a uniaxially-fixed mechanical boundary condition is one of the factors that determines the osteocyte alignment inside the bone matrix.
url https://doi.org/10.1038/s41598-021-88505-z
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