The effect of stiffness variation of electrospun fiber meshes of multiblock copolymers on the osteogenic differentiation of human mesenchymal stem cells

Abstract

Electrospinning has attracted significant attention as a method to produce cell culture substrates whose fibrous structure mimics the native extracellular matrix (ECM). In this study, the influence of E-modulus of fibrous substrates on the lineage commitment of human adipose-derived stem cells (hADSCs) was studied using fiber meshes prepared via the electrospinning of a polyetheresterurethane (PEEU) consisting of poly(ρ-dioxanone) (PPDO) and poly(ɛ-caprolactone) (PCL) segments. The PPDO: PCL weight ratio was varied from 40:60 to 70:30 to adjust the physiochemical properties of the PEEU fibers. The cells attached on stiffer PEEU70 (PPDO:PCL,= 70:30) fiber meshes displayed an elongated morphology compared to those cultured on softer fibers. The nuclear aspect ratio (width vs. length of a nucleus) of hADSCs cultured on softer PEEU40 (PPDO:PCL = 40:60) fibers was lower than on stiffer fibers. The osteogenic differentiation of hADSCs was enhanced by culturing on stiffer fibers. Compared to PEEU40, a 73% increase of osteocalcin expression and a 34% enhancement of alkaline phosphatase (ALP) activity was observed in cells on PEEU70. These results demonstrated that the differentiation commitment of stem cells could be regulated via tailoring the mechanical properties of electrospun fibers.

Keywords

Electrospun fibers polyetheresterurethane E-modulus mesenchymal stem cells osteogenic differentiation

Get full access to this article

View all access options for this article.

References

Jin

Y-Z

, Lee

. Mesenchymal Stem Cell Therapy for Bone Regeneration. Clinics in Orthopedic Surgery. 2018;10(3):271–8.

Zomorodian

, Baghaban Eslaminejad

. Mesenchymal Stem Cells as a Potent Cell Source for Bone Regeneration. Stem Cells International. 2012;2012:9.

Grayson

, et al.Stromal cells and stem cells in clinical bone regeneration. Nature Reviews Endocrinology. 2015;11:140.

James

. Review of Signaling Pathways Governing MSC Osteogenic and Adipogenic Differentiation. Scientifica (Cairo). 2013;2013:684736.

James

, et al.Perivascular stem cells: A prospectively purified mesenchymal stem cell population for bone tissue engineering. Stem Cells Transl Med. 2012;1(6):510–9.

Mizuno

, Tobita

, Uysal

. Concise review: Adipose-derived stem cells as a novel tool for future regenerative medicine. Stem Cells. 2012;30(5):804–10.

Levi

, Longaker

. Concise review: Adipose-derived stromal cells for skeletal regenerative medicine. Stem Cells. 2011;29(4):576–82.

Chamberlain

, et al.Concise review: Mesenchymal stem cells: Their phenotype, differentiation capacity, immunological features, and potential for homing. Stem Cells. 2007;25(11):2739–49.

Sun

, et al.Extracellular matrix stiffness controls osteogenic differentiation of mesenchymal stem cells mediated by integrin α5. Stem Cell Research & Therapy. 2018;9(1):52–2.

10.

Levi

, Longaker

. Concise Review: Adipose-Derived Stromal Cells for Skeletal Regenerative Medicine. STEM CELLS. 2011;29(4):576–82.

11.

Brett

, et al.Human Adipose-Derived Stromal Cell Isolation Methods and Use in Osteogenic and Adipogenic In Vivo Applications. Current Protocols in Stem Cell Biology. 2017;43:2H.1.1-2H.1.15.

12.

Grottkau

, Lin

. Osteogenesis of Adipose-Derived Stem Cells. Bone Research. 2013;1:133.

13.

O’Brien

. Biomaterials & scaffolds for tissue engineering. Materials Today. 2011;14(3):88–95.

14.

Bružauskaitė

, et al.Scaffolds and cells for tissue regeneration: Different scaffold pore sizes-different cell effects. Cytotechnology. 2016;68(3):355–69.

15.

Deitzel

, et al.Controlled deposition of electrospun poly(ethylene oxide) fibers. Polymer. 2001;42(19):8163–70.

16.

, Xia

. Electrospinning of Nanofibers: Reinventing the Wheel?Advanced Materials. 2004;16(14):1151–70.

17.

Min

, et al.Electrospinning of silk fibroin nanofibers and its effect on the adhesion and spreading of normal human keratinocytes and fibroblasts in vitro. Biomaterials. 2004;25(7-8):1289–97.

18.

Yoshimoto

, et al.A biodegradable nanofiber scaffold by electrospinning and its potential for bone tissue engineering. Biomaterials. 2003;24(12):2077–82.

19.

Porter

, Henson

, Popat

. Biodegradable poly(epsilon-caprolactone) nanowires for bone tissue engineering applications. Biomaterials. 2009;30(5):780–8.

20.

Wang

, et al.Synthesis of PDON-b-PEG-b-PDON block copolymers and drug delivery system thereof. Journal of Applied Polymer Science. 1998;68(13):2121–8.

21.

Kratz

, et al.Shape-Memory Properties and Degradation Behavior of Multifunctional Electro-Spun Scaffolds. The International Journal of Artificial Organs. 2011;34(2):225–30.

22.

, et al.Controlling major cellular processes of human mesenchymal stem cells using microwell structures. Adv Healthc Mater. 2014;3(12):1991–2003.

23.

Olivares-Navarrete

, et al.Substrate Stiffness Controls Osteoblastic and Chondrocytic Differentiation of Mesenchymal Stem Cells without Exogenous Stimuli. PLoS One. 2017;12(1):e0170312.

24.

Chancellor

, et al.Actomyosin tension exerted on the nucleus through nesprin-1 connections influences endothelial cell adhesion, migration, and cyclic strain-induced reorientation. Biophys J. 2010;99(1):115–23.

25.

Lombardi

, et al.The interaction between nesprins and sun proteins at the nuclear envelope is critical for force transmission between the nucleus and cytoskeleton. J Biol Chem. 2011;286(30):26743–53.

26.

Maniotis

, Chen

, Ingber

. Demonstration of mechanical connections between integrins, cytoskeletal filaments, and nucleoplasm that stabilize nuclear structure. Proc Natl Acad Sci U S A. 1997;94(3):849–54.

27.

Lovett

, et al.Modulation of Nuclear Shape by Substrate Rigidity. Cell Mol Bioeng. 2013;6(2):230–8.

28.

Dahl

, Ribeiro

, Lammerding

. Nuclear shape, mechanics, and mechanotransduction. Circ Res. 2008;102(11):1307–18.

29.

Bitar

, et al.Effect of cell density on osteoblastic differentiation and matrix degradation of biomimetic dense collagen scaffolds. Biomacromolecules. 2008;9(1):129–35.

30.

, et al.Effect of cell density on adipogenic differentiation of mesenchymal stem cells. Biochem Biophys Res Commun. 2009;381(3):322–7.

31.

McBeath

, et al.Cell shape, cytoskeletal tension, and RhoA regulate stem cell lineage commitment. Dev Cell. 2004;6(4):483–95.

32.

Jensen

, Gopalakrishnan

, Westendorf

. Regulation of gene expression in osteoblasts. Biofactors. 2010;36(1):25–32.