title: Modulus-driven differentiation of marrow stromal cells in 3D scaffolds that is independent of myosin-based cytoskeletal tension.
abstract: Proliferation and differentiation of cells are known to be influenced by the physical properties of the extracellular environment. Previous studies examining biophysics underlying cell response to matrix stiffness utilized a two-dimensional (2D) culture format, which is not representative of the three-dimensional (3D) tissue environment in vivo. We report on the effect of 3D matrix modulus on human bone marrow stromal cell (hBMSC) differentiation. hBMSCs underwent osteogenic differentiation in poly(ethylene glycol) hydrogels of all modulus (300-fold modulus range, from 0.2 kPa to 59 kPa) in the absence of osteogenic differentiation supplements. This osteogenic differentiation was modulus-dependent and was enhanced in stiffer gels. Osteogenesis in these matrices required integrin-protein ligation since osteogenesis was inhibited by soluble Arginine-Glycine-Aspartate-Serine peptide, which blocks integrin receptors. Immunostained images revealed lack of well-defined actin filaments and microtubules in the encapsulated cells. Disruption of mechanosensing elements downstream of integrin binding that have been identified from 2D culture such as actin filaments, myosin II contraction, and RhoA kinase did not abrogate hBMSC material-driven osteogenic differentiation in 3D. These data show that increased hydrogel modulus enhanced osteogenic differentiation of hBMSCs in 3D scaffolds but that hBMSCs did not use the same mechanosensing pathways that have been identified in 2D culture.
authors.affiliation: Polymers Division, National Institute of Standards and Technology, 100 Bureau Drive, Gaithersburg, MD 20899-8543, USA.
authors.forename: Sapun H
authors.initials: SH
authors.lastname: Parekh
authors.forename: Kaushik
authors.initials: K
authors.lastname: Chatterjee
authors.forename: Sheng
authors.initials: S
authors.lastname: Lin-Gibson
authors.forename: Nicole M
authors.initials: NM
authors.lastname: Moore
authors.forename: Marcus T
authors.initials: MT
authors.lastname: Cicerone
authors.forename: Marian F
authors.initials: MF
authors.lastname: Young
authors.forename: Carl G
authors.initials: CG
authors.lastname: Simon
mesh_terms: D000328:Adult; D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects; D003599:Cytoskeleton / Q000187:drug effects / Q000378:metabolism*; D055119:Elastic Modulus / Q000187:drug effects; D005260:Female; D005353:Fibronectins / Q000494:pharmacology; D006801:Humans; D008689:Methacrylates / Q000737:chemistry; D051379:Mice; D009218:Myosins / Q000378:metabolism*; D009842:Oligopeptides / Q000494:pharmacology; D010012:Osteogenesis / Q000187:drug effects; D011092:Polyethylene Glycols / Q000737:chemistry; D017154:Stromal Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D054457:Tissue Scaffolds / Q000737:chemistry*
chemical_list: D005353:Fibronectins; D008689:Methacrylates; D009842:Oligopeptides; D011092:Polyethylene Glycols; C046617:arginyl-glycyl-aspartyl-serine; D009218:Myosins