Unique osteogenic profile of bone marrow stem cells stimulated in perfusion bioreactor is Rho-ROCK-mediated contractility dependent.
Rho GTPase signaling
actomyosin contraction
bioreactor
bone tissue engineering
fluid shear stress
osteogenic differentiation
Journal
Bioengineering & translational medicine
ISSN: 2380-6761
Titre abrégé: Bioeng Transl Med
Pays: United States
ID NLM: 101689146
Informations de publication
Date de publication:
May 2023
May 2023
Historique:
received:
21
12
2022
revised:
28
02
2023
accepted:
04
03
2023
medline:
19
5
2023
pubmed:
19
5
2023
entrez:
19
5
2023
Statut:
epublish
Résumé
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.
Identifiants
pubmed: 37206242
doi: 10.1002/btm2.10509
pii: BTM210509
pmc: PMC10189446
doi:
Types de publication
Journal Article
Langues
eng
Pagination
e10509Informations de copyright
© 2023 The Authors. Bioengineering & Translational Medicine published by Wiley Periodicals LLC on behalf of American Institute of Chemical Engineers.
Déclaration de conflit d'intérêts
The authors declare no potential conflicts of interest with respect to the authorship and/or publication of this article.
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