Electrically conducting polymers are prospective candidates as active substrates for the development of neuroprosthetic devices. The utility of these substrates for promoting differentiation of embryonic stem cells paves viable routes for regenerative medicine. Here, we have tuned the electrical and mechanical cues provided to the embryonic stem cells during differentiation by precisely straining the conducting polymer (CP) coated, elastomeric-substrate. Upon straining the substrates, the neural differentiation pattern occurs in form of aggregates, accompanied by a gradient where substrate interface reveals a higher degree of differentiation. The CP domains align under linear stress along with the formation of local defect patterns leading to disruption of actin cytoskeleton of cells, and can provide a mechano-transductive basis for the observed changes in the differentiation. Our results demonstrate that along with biochemical and mechanical cues, conductivity of the polymer plays a major role in cellular differentiation thereby providing another control feature to modulate the differentiation and proliferation of stem cells.
Get full access to this article
View all access options for this article.
References
1.
MeredithJ.E., FazeliB., SchwartzM.A.The extracellular-matrix as a cell-survival factor. Mol Biol Cell, 4:953. 1993.
GumbinerB.M.Cell adhesion: the molecular basis of tissue architecture and morphogenesis. Cell, 84:345. 1996.
4.
BurridgeK., FathK., KellyT., NuckollsG., TurnerC.Focal adhesions: transmembrane junctions between the extracellular matrix and the cytoskeleton. Annu Rev Cell Biol, 4:487. 1988.
5.
DemboM., WangY.L.Stresses at the cell-to-substrate interface during locomotion of fibroblasts. Biophys J, 76:2307. 1999.
6.
HallidayN.L., TomasekJ.J.Mechanical properties of the extracellular matrix influence fibronectin fibril assembly in vitro. Exp Cell Res, 217:109. 1995.
7.
YimE.K.F., DarlingE.M., KulangaraK., GuilakF., LeongK.W.Nanotopography-induced changes in focal adhesions, cytoskeletal organization, and mechanical properties of human mesenchymal stem cells. Biomaterials, 31:1299. 2010.
8.
HuebschN., AranyP.R., MaoA.S., ShvartsmanD., AliO.A., BencherifS.A.et al.Harnessing traction-mediated manipulation of the cell/matrix interface to control stem-cell fate. Nat Mater, 9:518. 2010.
9.
ParekhS.H., ChatterjeeK., Lin-GibsonS., MooreN.M., CiceroneM.T., YoungM.F.et al.Modulus-driven differentiation of marrow stromal cells in 3D scaffolds that is independent of myosin-based cytoskeletal tension. Biomaterials, 32:2256. 2011.
10.
BalabanN.Q.Force and focal adhesion assembly: a close relationship studied using elastic micropatterned substrates. Nat Cell Biol, 3:466. 2001.
11.
BeningoK.A., DemboM., KaverinaI., SmallJ.V., WangY.L.Nascent focal adhesions are responsible for the generation of strong propulsive forces in migrating fibroblasts. J Cell Biol, 153:881. 2001.
CurtisA., WilkinsonC.New depths in cell behaviour: reactions of cells to nanotopography. Biochem Soc Symp, 65:15. 1999.
14.
RowlandsA.S., GeorgeP.A., Cooper-WhiteJ.J.Directing osteogenic and myogenic differentiation of MSCs: interplay of stiffness and adhesive ligand presentation. Am J Physiol Cell Physiol, 295:C1037. 2008.
15.
WingateK., BonaniW., TanY., BryantS.J., TanW.Compressive elasticity of three-dimensional nanofiber matrix directs mesenchymal stem cell differentiation to vascular cells with endothelial or smooth muscle cell markers. Acta Biomater, 8:1440. 2012.
PekY.S., WanA.C.A., YingJ.Y.The effect of matrix stiffness on mesenchymal stem cell differentiation in a 3D thixotropic gel. Biomaterials, 31:385. 2010.
18.
SchmidtC.E., ShastriV.R., VacantiJ.P., LangerR.Stimulation of neurite outgrowth using an electrically conducting polymer. Proc Natl Acad Sci USA, 94:8948. 1997.
19.
BolinM.H., SvennerstenK., WangX.J., ChronakisI.S., Richter-DahlforsA., JagerE.W.H.et al.Nano-fiber scaffold electrodes based on PEDOT for cell stimulation. Sens Actuators B Chem, 142:451. 2009.
20.
Del ValleL.J., AradillaD., OliverR., SepulcreF., GamezA., ArmelinE.et al.Cellular adhesion and proliferation on poly(3,4-ethylenedioxythiophene): benefits in the electroactivity of the conducting polymer. Eur Polym J, 43:2342. 2007.
21.
GomezN., LeeJ.Y., NickelsJ.D., SchmidtC.E.Micropatterned polypyrrole: a combination of electrical and topographical characteristics for the stimulation of cells. Adv Funct Mater, 17:1645. 2007.
AbidianM.R., LudwigK.A., MarzulloT.C., MartinD.C., KipkeD.R.Interfacing conducting polymer nanotubes with the central nervous system: chronic neural recording using poly(3,4-ethylenedioxythiophene) nanotubes. Adv Mater, 2:3764. 2009.
24.
XiaoY.H., MartinD.C., CuiX.Y., ShenaiM.Surface modification of neural probes with conducting polymer poly(hydroxymethylated-3,4-ethylenedioxythiophene) and its biocompatibility. Appl Biochem Biotechnol, 128:117. 2006.
25.
KimD.H., LuN.S., GhaffariR., KimY.S., LeeS.P., XuL.Z.et al.Materials for multifunctional balloon catheters with capabilities in cardiac electrophysiological mapping and ablation therapy. Nat Mater, 10:316. 2011.
26.
GeorgeP.M., LyckmanA.W., LaVanD.A., HegdeA., LeungY., AvasareR.et al.Fabrication and biocompatibility of polypyrrole implants suitable for neural prosthetics. Biomaterials, 26:3511. 2005.
27.
KimD.H., Richardson-BurnsS.M., HendricksJ.L., SequeraC., MartinD.C.Effect of immobilized nerve growth factor on conductive polymers: electrical properties and cellular response. Adv Funct Mater, 17:79. 2007.
28.
ChristophersonG.T., SongH., MaoH-Q.The influence of fiber diameter of electrospun substrates on neural stem cell differentiation and proliferation. Biomaterials, 30:556. 2009.
29.
RecknorJ.B., SakaguchiD.S., MallapragadaS.K.Directed growth and selective differentiation of neural progenitor cells on micropatterned polymer substrates. Biomaterials, 27:4098. 2006.
30.
GeorgesP.C., MillerW.J., MeaneyD.F., SawyerE.S., JanmeyP.A.Matrices with compliance comparable to that of brain tissue select neuronal over glial growth in mixed cortical cultures. Biophys J, 90:3012. 2006.
TeixeiraA.I., IlkhanizadehS., WigeniusJ.A., DuckworthJ.K., InganasO., HermansonO.The promotion of neuronal maturation on soft substrates. Biomaterials, 30:4567. 2009.
33.
AyalaR., ZhangC., YangD., HwangY., AungA., ShroffS.S.et al.Engineering the cell–material interface for controlling stem cell adhesion, migration, and differentiation. Biomaterials, 32:3700. 2011.
34.
KotwalA., SchmidtC.E.Electrical stimulation alters protein adsorption and nerve cell interactions with electrically conducting biomaterials. Biomaterials, 22:1055. 2001.
35.
WongJ.Y., LangerR., IngberD.E.Electrically conducting polymers can noninvasively control the shape and growth of mammalian cells. Proc Natl Acad Sci USA, 91:3201. 1994.
36.
LordM.S., FossM., BesenbacherF.Influence of nanoscale surface topography on protein adsorption and cellular response. Nano Today, 5:66. 2010.
37.
VijayV., RaoA.D., NarayanK.S.In situ studies of strain dependent transport properties of conducting polymers on elastomeric substrates. J Appl Phys, 109:084525. 2011.
38.
BainG., KitchensD., YaoM., HuettnerJ.E., GottliebD.I.Embryonic stem cells express neuronal properties in vitro. Dev Biol, 168:342. 1995.
39.
JagathaB., DivyaM.S., SanalkumarR., IndulekhaC.L., VidyanandS., DivyaT.S.et al.In vitro differentiation of retinal ganglion-like cells from embryonic stem cell derived neural progenitors. Biochem Biophys Res Commun, 380:230. 2009.
40.
KaufmanD.L., HouserC.R., TobinA.J.Two forms of the gamma-aminobutyric acid synthetic enzyme glutamate decarboxylase have distinct intraneuronal distributions and cofactor interactions. J Neurochem, 56:720. 1991.
41.
GeppertM., GodaY., HammerR.E., LiC., RosahlT.W., StevensC.F.et al.Synaptotagmin I: a major Ca2+ sensor for transmitter release at a central synapse. Cell, 79:717. 1994.
42.
LongairM.H., BakerD.A., ArmstrongJ.D.Simple Neurite Tracer: open source software for reconstruction, visualization and analysis of neuronal processes. Bioinformatics, 27:2453. 2011.
43.
AbramoffM.D., MagelhaesP.J., RamS.J.Image processing with ImageJ. Biophotonics Int, 11:36. 2004.
44.
SchneiderC.A., RasbandW.S., EliceiriK.W.NIH Image to ImageJ: 25 years of image analysis. Nat Methods, 9:671. 2012.
45.
ZhuX., MillsK.L., PetersP.R., BahngJ.H, LiuE.H., ShimJ.et al.Fabrication of reconfigurable protein matrices by cracking. Nat Mater, 4:403. 2005.
46.
LipomiD.J., LeeJ.A., VosgueritchianM., TeeB.C.K., BolanderJ.A., BaoZ.A.Electronic properties of transparent conductive films of PEDOT:PSS on stretchable substrates. Chem Mater, 24:373. 2012.
47.
McBeathR., PironeD.M., NelsonC.M., BhadrirajuK., ChenC.S.Cell shape, cytoskeletal tension, and rhoa regulate stem cell lineage commitment. Dev Cell, 6:483. 2004.
48.
DischerD.E., JanmeyP., WangY.L.Tissue cells feel and respond to the stiffness of their substrate. Science, 310:1139. 2005.
49.
EyckmansJ., BoudouT., YuX., ChenC.S.A Hitchhiker's guide to mechanobiology. Dev Cell, 21:35. 2011.
50.
PoonB.C., DiasP., AnsemsP., ChumS.P., HiltnerA., BaerE.Structure and deformation of an elastomeric propylene–ethylene copolymer. J Appl Polym Sci, 10:489. 2007.
Supplementary Material
Please find the following supplemental material available below.
For Open Access articles published under a Creative Commons License, all supplemental material carries the same license as the article it is associated with.
For non-Open Access articles published, all supplemental material carries a non-exclusive license, and permission requests for re-use of supplemental material or any part of supplemental material shall be sent directly to the copyright owner as specified in the copyright notice associated with the article.
