The basement membrane (BM) of the corneal epithelium presents biophysical cues in the form of topography and compliance that can impact the phenotype and behaviors of cells and their nuclei through modulation of cytoskeletal dynamics. In addition, it is also well known that the intrinsic biochemical attributes of BMs can modulate cell behaviors. In this study, the influence of the combination of exogenous coating of extracellular matrix proteins (ECM) (fibronectin-collagen [FNC]) with substratum topography was investigated on cytoskeletal architecture as well as alignment and migration of immortalized corneal epithelial cells. In the absence of FNC coating, a significantly greater percentage of cells aligned parallel with the long axis of the underlying anisotropically ordered topographic features; however, their ability to migrate was impaired. Additionally, changes in the surface area, elongation, and orientation of cytoskeletal elements were differentially influenced by the presence or absence of FNC. These results suggest that the effects of topographic cues on cells are modulated by the presence of surface-associated ECM proteins. These findings have relevance to experiments using cell cultureware with biomimetic biophysical attributes as well as the integration of biophysical cues in tissue-engineering strategies and the development of improved prosthetics.
Get full access to this article
View all access options for this article.
References
1.
AbramsG.A., GoodmanS.L., NealeyP.F., FrancoM., MurphyC.J.Nanoscale topography of the basement membrane underlying the corneal epithelium of the rhesus macaque. Cell Tissue Res, 299:39. 2000.
2.
AbramsG.A., SchausS.S., GoodmanS.L., NealeyP.F., MurphyC.J.Nanoscale topography of the corneal epithelial basement Membrane and Descemet's membrane of the human. Cornea, 19:57. 2000.
3.
KaruriN.W., PorriT.J., AlbrechtR.M., MurphyC.J., NealeyP.F.Nano- and microscale holes modulate cell-substrate adhesion, cytoskeletal organization, and -beta1 integrin localization in SV40 human corneal epithelial cells. IEEE Trans Nanobioscience, 5:273. 2006.
KaruriN.W., NealeyP.F., MurphyC.J., AlbrechtR.M.Structural organization of the cytoskeleton in SV40 human corneal epithelial cells cultured on nano- and microscale grooves. Scanning, 30:405. 2008.
6.
LiliensiekS.J., WoodJ.A., YongJ., AuerbachR., NealeyP.F., MurphyC.J.Modulation of human vascular endothelial cell behaviors by nanotopographic cues. Biomaterials, 31:5418. 2010.
7.
PotS.A., LiliensiekS.J., MyrnaK.E., BentleyE., JesterJ.V., NealeyP.F., MurphyC.J.Nanoscale topography–induced modulation of fundamental cell behaviors of rabbit corneal keratocytes, fibroblasts, and myofibroblasts. Invest Ophthalmol Vis Sci, 51:1373. 2010.
8.
TocceE.J., SmirnovV.K., KibalovD.S., LiliensiekS.J., MurphyC.J., NealeyP.F.The ability of corneal epithelial cells to recognize high aspect ratio nanostructures. Biomaterials, 31:4064. 2010.
9.
DalbyM.J., RiehleM.O., YarwoodS.J., WilkinsonC.D.W., CurtisA.S.G.Nucleus alignment and cell signaling in fibroblasts: response to a micro-grooved topography. Exp Cell Res, 284:272. 2003.
10.
McKee ClaytonT., Raghunathan VijayK., Nealey PaulF., RussellP., Murphy ChristopherJ.Topographic modulation of the orientation and shape of cell nuclei and their influence on the measured elastic modulus of epithelial cells. Biophys J, 101:2139. 2011.
11.
MorganJ.T., WoodJ.A., ShahN.M., HughbanksM.L., RussellP., BarakatA.I., MurphyC.J.Integration of basal topographic cues and apical shear stress in vascular endothelial cells. Biomaterials, 33:4126. 2012.
12.
FoleyJ.D., GrunwaldE.W., NealeyP.F., MurphyC.J.Cooperative modulation of neuritogenesis by PC12 cells by topography and nerve growth factor. Biomaterials, 26:3639. 2005.
13.
ThomasyS.M., WoodJ.A., KassP.H., MurphyC.J., RussellP.Substratum stiffness and latrunculin B regulate matrix gene and protein expression in human trabecular meshwork cells. Invest Ophthalmol Vis Sci, 53:952. 2012.
14.
WoodJ.A., McKeeC.T., ThomasyS.M., FischerM.E., ShahN.M., MurphyC.J., RussellP.Substratum compliance regulates human trabecular meshwork cell behaviors and response to latrunculin B. Invest Ophthalmol Vis Sci, 52:9298. 2011.
15.
TeixeiraA.I., McKieG.A., FoleyJ.D., BerticsP.J., NealeyP.F., MurphyC.J.The effect of environmental factors on the response of human corneal epithelial cells to nanoscale substrate topography. Biomaterials, 27:3945. 2006.
16.
KooS., AhnS., ZhangH., WangJ., YimE.Human corneal keratocyte response to micro- and nano-gratings on chitosan and PDMS. Cell Mol Bioeng, 4:399. 2011.
17.
WangG., LiuX., ZreiqatH., DingC.Enhanced effects of nano-scale topography on the bioactivity and osteoblast behaviors of micron rough ZrO2 coatings. Colloids and Surfaces B: Biointerfaces, 86:267. 2011.
18.
SweeneyD.F., XieR.Z., EvansM.D.M., VannasA., ToutS.D., GriesserH.J., JohnsonG., SteeleJ.G.A comparison of biological coatings for the promotion of corneal epithelialization of synthetic surface in vivo. Invest Ophthalmol Vis Sci, 44:3301. 2003.
19.
TocceE.J., BroderickA.H., MurphyK.C., LiliensiekS.J., MurphyC.J., LynnD.M., NealeyP.F.Functionalization of reactive polymer multilayers with RGD and an antifouling motif: RGD density provides control over human corneal epithelial cell–substrate interactions. J Biomed Mater Res Part A, 100A:84. 2012.
20.
GrohmannS., RotheH., FrantM., LiefeithK.Colloidal force spectroscopy and cell biological investigations on biomimetic polyelectrolyte multilayer coatings composed of chondroitin sulfate and heparin. Biomacromolecules, 12:1987. 2011.
21.
van HorssenR., GaljartN., RensJ.A.P., EggermontA.M.M., ten HagenT.L.M.Differential effects of matrix and growth factors on endothelial and fibroblast motility: application of a modified cell migration assay. J Cell Biochem, 99:1536. 2006.
22.
AucoinL., GriffithC.M., PleizierG., DeslandesY., SheardownH.Interactions of corneal epithelial cells and surfaces modified with cell adhesion peptide combinations. J Biomater Sci Polym Ed, 13:447. 2002.
23.
FongE., TzlilS., TirrellD.A.Boundary crossing in epithelial wound healing. Proc Natl Acad Sci U S A, 107:19302. 2010.
24.
HuangZ., SargeantT.D., HulvatJ.F., MataA., BringasP.Jr., KohC.Y., StuppS.I., SneadM.L.Bioactive nanofibers instruct cells to proliferate and differentiate during enamel regeneration. J Bone Miner Res, 23:1995. 2008.
25.
LeeK.Y., AlsbergE., HsiongS., ComisarW., LindermanJ., ZiffR., MooneyD.Nanoscale adhesion ligand organization regulates osteoblast proliferation and differentiation. Nano Lett, 4:1501. 2004.
26.
RowleyJ.A., MooneyD.J.Alginate type and RGD density control myoblast phenotype. J Biomed Mater Res, 60:217. 2002.
27.
WangH., MaL., YangS., ShaoZ., MengC., DuanD., LiY.Effect of RGD-modified silk material on the adhesion and proliferation of bone marrow-derived mesenchymal stem cells. J Huazhong Univ Sci Technolog Med Sci, 29:80. 2009.
28.
SchwartzM.A.Integrins and extracellular matrix in mechanotransduction. Cold Spring Harbor Perspect Biol, 2:a005066. 2010.
29.
SchwartzM.A., AssoianR.K.Integrins and cell proliferation. J Cell Sci, 114:2553. 2001.
30.
MyrnaK.E., PotS.A., MurphyC.J.Meet the corneal myofibroblast: the role of myofibroblast transformation in corneal wound healing and pathology. Vet Ophthalmol, 12:25. 2009.
31.
TeixeiraA.I., AbramsG.A., BerticsP.J., MurphyC.J., NealeyP.F.Epithelial contact guidance on well-defined micro- and nanostructured substrates. J Cell Sci, 116:1881. 2003.
32.
OdomT.W., LoveJ.C., WolfeD.B., PaulK.E., WhitesidesG.M.Improved pattern transfer in soft lithography using composite stamps. Langmuir, 18:5314. 2002.
33.
RobertsonD.M., LiL., FisherS., PearceV.P., ShayJ.W., WrightW.E., CavanaghH.D., JesterJ.V.Characterization of growth and differentiation in a telomerase-immortalized human corneal epithelial cell line. Invest Ophthalmol Vis Sci, 46:470. 2005.
34.
KabosovaA., AzarD.T., BannikovG.A., CampbellK.P., DurbeejM., GhohestaniR.F., JonesJ.C.R., KenneyM.C., KochM., NinomiyaY.Compositional differences between infant and adult human corneal basement membranes. Invest Ophthalmol Vis Sci, 48:4989. 2007.
35.
LjubimovA.V., BurgesonR.E., ButkowskiR.J., MichaelA.F., SunT.T., KenneyM.C.Human corneal basement membrane heterogeneity: topographical differences in the expression of type IV collagen and laminin isoforms. Lab Invest, 72:461. 1995.
36.
MadriJ.A., MarxM.Matrix composition, organization and soluble factors: modulators of microvascular cell differentiation in vitro. Kidney Int, 41:560. 1992.
37.
MadriJ.A., MarxM., MerwinJ.R., BassonC., PrinzC., BellL.Effects of soluble factors and extracellular matrix components on vascular cell behavior in vitro and in vivo: models of de-endothelialization and repair. J Cell Biochem, 45:123. 1991.
38.
JulianoR.Cooperation between soluble factors and integrin-mediated cell anchorage in the control of cell growth and differentiation. Bioessays, 18:911. 1996.
39.
FraserS.A., TingY.-H., MallonK.S., WendtA.E., MurphyC.J., NealeyP.F.Sub-micron and nanoscale feature depth modulates alignment of stromal fibroblasts and corneal epithelial cells in serum-rich and serum-free media. J Biomed Mater Res Part A, 86A:725. 2008.
40.
HealyK.E., ThomasC.H., RezaniaA., KimJ.E., McKeownP.J., LomB., HockbergerP.E.Kinetics of bone cell organization and mineralization on materials with patterned surface chemistry. Biomaterials, 17:195. 1996.
41.
AnselmeK., LinezP., BigerelleM., Le MaguerD., Le MaguerA., HardouinP., HildebrandH.F., IostA., LeroyJ.M.The relative influence of the topography and chemistry of TiAl6V4 surfaces on osteoblastic cell behaviour. Biomaterials, 21:1567. 2000.
StevensM.M., GeorgeJ.H.Exploring and engineering the cell surface interface. Science, 310:1135. 2005.
44.
MaroisY., Sugot-LuizardM.F., GuidoinR.Endothelial cell behavior on vascular prosthetic grafts: effect of polymer chemistry, surface structure, and surface treatment. ASAIO J, 45:272. 1999.
45.
PonsonnetL., ReybierK., JaffrezicN., ComteV., LagneauC., LissacM., MarteletC.Relationship between surface properties (roughness, wettability) of titanium and titanium alloys and cell behaviour. Mater Sci Eng C, 23:551. 2003.
46.
DeligianniD.D., KatsalaN.D., KoutsoukosP.G., MissirlisY.F.Effect of surface roughness of hydroxyapatite on human bone marrow cell adhesion, proliferation, differentiation and detachment strength. Biomaterials, 22:87. 2000.
47.
HallabN.J., BundyK.J., O'ConnorK., MosesR.L., JacobsJ.J.Evaluation of metallic and polymeric biomaterial surface energy and surface roughness characteristics for directed cell adhesion. Tissue Eng, 7:55. 2001.
48.
XuC., YangF., WangS., RamakrishnaS.In vitro study of human vascular endothelial cell function on materials with various surface roughness. J Biomed Mater Res Part A, 71A:154. 2004.
49.
HoutchensG.R., FosterM.D., DesaiT.A., MorganE.F., WongJ.Y.Combined effects of microtopography and cyclic strain on vascular smooth muscle cell orientation. J Biomech, 41:762. 2008.
50.
WilsonM.J., LiliensiekS.J., MurphyC.J., MurphyW.L., NealeyP.F.Hydrogels with well-defined peptide-hydrogel spacing and concentration: impact on epithelial cell behavior. Soft Matter, 8:390. 2012.
NishidaT., NakagawaS., AwataT., OhashiY., WatanabeK., ManabeR.Fibronectin promotes epithelial migration of cultured rabbit cornea in situ. J Cell Biol, 97:1653. 1983.
53.
WatanabeK., NakagawaS., NishidaT.Stimulatory effects of fibronectin and EGF on migration of corneal epithelial cells. Invest Ophthalmol Vis Sci, 28:205. 1987.
54.
DiehlK.A., FoleyJ.D., NealeyP.F., MurphyC.J.Nanoscale topography modulates corneal epithelial cell migration. J Biomed Mater Res Part A, 75A:603. 2005.
55.
RidleyA.J., SchwartzM.A., BurridgeK., FirtelR.A., GinsbergM.H., BorisyG., ParsonsJ.T., HorwitzA.R.Cell migration: integrating signals from front to back. Science, 302:1704. 2003.
56.
GasiorowskiJ.Z., LiliensiekS.J., RussellP., StephanD.A., NealeyP.F., MurphyC.J.Alterations in gene expression of human vascular endothelial cells associated with nanotopographic cues. Biomaterials, 31:8882. 2010.
57.
YimE.K.F., LeongK.W.Significance of synthetic nanostructures in dictating cellular response. Nanomedicine, 1:10. 2005.
58.
SchoenwaelderS.M., BurridgeK.Bidirectional signaling between the cytoskeleton and integrins. Curr Opin Cell Biol, 11:274. 1999.
HoshibaT., YamadaT., LuH., KawazoeN., TateishiT., ChenG.Nuclear deformation and expression change of cartilaginous genes during in vitro expansion of chondrocytes. Biochem Biophys Res Commun, 374:688. 2008.
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.
