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Unique osteogenic profile of bone marrow stem cells stimulated in perfusion bioreactor is Rho‐ROCK‐mediated contractility dependent

The fate determination of bone marrow mesenchymal stem/stromal cells (BMSC) is tightly regulated by mechanical cues, including fluid shear stress. Knowledge of mechanobiology in 2D culture has allowed researchers in bone tissue engineering to develop 3D dynamic culture systems with the potential for...

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Autores principales: Yamada, Shuntaro, Yassin, Mohammed A., Torelli, Francesco, Hansmann, Jan, Green, Jeremy B. A., Schwarz, Thomas, Mustafa, Kamal
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley & Sons, Inc. 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10189446/
https://www.ncbi.nlm.nih.gov/pubmed/37206242
http://dx.doi.org/10.1002/btm2.10509
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author Yamada, Shuntaro
Yassin, Mohammed A.
Torelli, Francesco
Hansmann, Jan
Green, Jeremy B. A.
Schwarz, Thomas
Mustafa, Kamal
author_facet Yamada, Shuntaro
Yassin, Mohammed A.
Torelli, Francesco
Hansmann, Jan
Green, Jeremy B. A.
Schwarz, Thomas
Mustafa, Kamal
author_sort Yamada, Shuntaro
collection PubMed
description The fate determination of bone marrow mesenchymal stem/stromal cells (BMSC) is tightly regulated by mechanical cues, including fluid shear stress. Knowledge of mechanobiology in 2D culture has allowed researchers in bone tissue engineering to develop 3D dynamic culture systems with the potential for clinical translation in which the fate and growth of BMSC are mechanically controlled. However, due to the complexity of 3D dynamic cell culture compared to the 2D counterpart, the mechanisms of cell regulation in the dynamic environment remain relatively undescribed. In the present study, we analyzed the cytoskeletal modulation and osteogenic profiles of BMSC under fluid stimuli in a 3D culture condition using a perfusion bioreactor. BMSC subjected to fluid shear stress (mean 1.56 mPa) showed increased actomyosin contractility, accompanied by the upregulation of mechanoreceptors, focal adhesions, and Rho GTPase‐mediated signaling molecules. Osteogenic gene expression profiling revealed that fluid shear stress promoted the expression of osteogenic markers differently from chemically induced osteogenesis. Osteogenic marker mRNA expression, type 1 collagen formation, ALP activity, and mineralization were promoted in the dynamic condition, even in the absence of chemical supplementation. The inhibition of cell contractility under flow by Rhosin chloride, Y27632, MLCK inhibitor peptide‐18, or Blebbistatin revealed that actomyosin contractility was required for maintaining the proliferative status and mechanically induced osteogenic differentiation in the dynamic culture. The study highlights the cytoskeletal response and unique osteogenic profile of BMSC in this type of dynamic cell culture, stepping toward the clinical translation of mechanically stimulated BMCS for bone regeneration.
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spelling pubmed-101894462023-05-18 Unique osteogenic profile of bone marrow stem cells stimulated in perfusion bioreactor is Rho‐ROCK‐mediated contractility dependent Yamada, Shuntaro Yassin, Mohammed A. Torelli, Francesco Hansmann, Jan Green, Jeremy B. A. Schwarz, Thomas Mustafa, Kamal Bioeng Transl Med Research Articles The fate determination of bone marrow mesenchymal stem/stromal cells (BMSC) is tightly regulated by mechanical cues, including fluid shear stress. Knowledge of mechanobiology in 2D culture has allowed researchers in bone tissue engineering to develop 3D dynamic culture systems with the potential for clinical translation in which the fate and growth of BMSC are mechanically controlled. However, due to the complexity of 3D dynamic cell culture compared to the 2D counterpart, the mechanisms of cell regulation in the dynamic environment remain relatively undescribed. In the present study, we analyzed the cytoskeletal modulation and osteogenic profiles of BMSC under fluid stimuli in a 3D culture condition using a perfusion bioreactor. BMSC subjected to fluid shear stress (mean 1.56 mPa) showed increased actomyosin contractility, accompanied by the upregulation of mechanoreceptors, focal adhesions, and Rho GTPase‐mediated signaling molecules. Osteogenic gene expression profiling revealed that fluid shear stress promoted the expression of osteogenic markers differently from chemically induced osteogenesis. Osteogenic marker mRNA expression, type 1 collagen formation, ALP activity, and mineralization were promoted in the dynamic condition, even in the absence of chemical supplementation. The inhibition of cell contractility under flow by Rhosin chloride, Y27632, MLCK inhibitor peptide‐18, or Blebbistatin revealed that actomyosin contractility was required for maintaining the proliferative status and mechanically induced osteogenic differentiation in the dynamic culture. The study highlights the cytoskeletal response and unique osteogenic profile of BMSC in this type of dynamic cell culture, stepping toward the clinical translation of mechanically stimulated BMCS for bone regeneration. John Wiley & Sons, Inc. 2023-03-17 /pmc/articles/PMC10189446/ /pubmed/37206242 http://dx.doi.org/10.1002/btm2.10509 Text en © 2023 The Authors. Bioengineering & Translational Medicine published by Wiley Periodicals LLC on behalf of American Institute of Chemical Engineers. https://creativecommons.org/licenses/by/4.0/This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Articles
Yamada, Shuntaro
Yassin, Mohammed A.
Torelli, Francesco
Hansmann, Jan
Green, Jeremy B. A.
Schwarz, Thomas
Mustafa, Kamal
Unique osteogenic profile of bone marrow stem cells stimulated in perfusion bioreactor is Rho‐ROCK‐mediated contractility dependent
title Unique osteogenic profile of bone marrow stem cells stimulated in perfusion bioreactor is Rho‐ROCK‐mediated contractility dependent
title_full Unique osteogenic profile of bone marrow stem cells stimulated in perfusion bioreactor is Rho‐ROCK‐mediated contractility dependent
title_fullStr Unique osteogenic profile of bone marrow stem cells stimulated in perfusion bioreactor is Rho‐ROCK‐mediated contractility dependent
title_full_unstemmed Unique osteogenic profile of bone marrow stem cells stimulated in perfusion bioreactor is Rho‐ROCK‐mediated contractility dependent
title_short Unique osteogenic profile of bone marrow stem cells stimulated in perfusion bioreactor is Rho‐ROCK‐mediated contractility dependent
title_sort unique osteogenic profile of bone marrow stem cells stimulated in perfusion bioreactor is rho‐rock‐mediated contractility dependent
topic Research Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10189446/
https://www.ncbi.nlm.nih.gov/pubmed/37206242
http://dx.doi.org/10.1002/btm2.10509
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