{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,15]],"date-time":"2026-09-15T21:20:27Z","timestamp":1789507227339,"version":"build-2803163510"},"reference-count":48,"publisher":"MDPI AG","issue":"24","license":[{"start":{"date-parts":[[2025,12,11]],"date-time":"2025-12-11T00:00:00Z","timestamp":1765411200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"crossref","award":["P30 EY030413"],"award-info":[{"award-number":["P30 EY030413"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"crossref","award":["R01 EY030190"],"award-info":[{"award-number":["R01 EY030190"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/100005931","name":"International Foundation for Ethical Research","doi-asserted-by":"crossref","id":[{"id":"10.13039\/100005931","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/100001818","name":"Research to Prevent Blindness","doi-asserted-by":"crossref","id":[{"id":"10.13039\/100001818","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Corneal fibroblasts adhere to the extracellular matrix via integrin-containing focal adhesions (FAs). Although topographical cues are known to influence FA patterning in corneal fibroblasts, it is unclear how ECM composition, biophysical cues, and specific integrins modulate FA patterning in corneal fibroblasts. In this study, we cultured a human corneal fibroblast cell line (HTKs) on different ECM proteins and micropatterns of aligned collagen fibrils to determine the effects of ECM topography and composition on focal adhesion subcellular patterning. Using confocal imaging, we observed and quantified changes in FA and integrin patterning based on the underlying ECM type. More specifically, the presence of fibrillar topography as compared to monomeric collagen resulted in diminished FA number, area, and length. Using specific integrin blocking antibodies, we also demonstrate that HTKs use different integrin subunits to adhere to specific ECM coatings. For example, \u03b21 integrins are important in adhesion formation when corneal fibroblasts adhere to collagen, while \u03b15 integrin is important for the HTKs to adhere to fibronectin. Blocking of \u03b15 integrin did not completely inhibit cell spreading and FA patterning when cells adhered to fibronectin. These results suggest that there might be other fibronectin receptors that HTKs use in the absence of \u03b15 integrin. These results lay the foundation to understand the role of different integrin subunits in FA patterning. Through further experimentation using our developed platform, we envision that a better understanding of the integrins and their associated signaling could have implications for advanced in vitro and in vivo applications in cornea biology.<\/jats:p>","DOI":"10.3390\/ijms262411935","type":"journal-article","created":{"date-parts":[[2025,12,11]],"date-time":"2025-12-11T11:17:27Z","timestamp":1765451847000},"page":"11935","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Effects of Topography and Extracellular Matrix Composition on Focal Adhesion Patterning in Human Corneal Fibroblasts"],"prefix":"10.3390","volume":"26","author":[{"given":"Divya","family":"Subramanian","sequence":"first","affiliation":[{"name":"Department of Bioengineering, University of Texas at Dallas, Richardson, TX 75080, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2485-6498","authenticated-orcid":false,"given":"Nathaniel S.","family":"Tjahjono","sequence":"additional","affiliation":[{"name":"Department of Bioengineering, University of Texas at Dallas, Richardson, TX 75080, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tarik Z.","family":"Shihabeddin","sequence":"additional","affiliation":[{"name":"Department of Bioengineering, University of Texas at Dallas, Richardson, TX 75080, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Satweka","family":"Nammi","sequence":"additional","affiliation":[{"name":"Department of Bioengineering, University of Texas at Dallas, Richardson, TX 75080, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Miguel","family":"Miron-Mendoza","sequence":"additional","affiliation":[{"name":"Department of Ophthalmology, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Victor D.","family":"Varner","sequence":"additional","affiliation":[{"name":"Department of Bioengineering, University of Texas at Dallas, Richardson, TX 75080, USA"},{"name":"Department of Biomedical Engineering, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0795-4539","authenticated-orcid":false,"given":"W. Matthew","family":"Petroll","sequence":"additional","affiliation":[{"name":"Department of Ophthalmology, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA"},{"name":"Department of Biomedical Engineering, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6404-318X","authenticated-orcid":false,"given":"David W.","family":"Schmidtke","sequence":"additional","affiliation":[{"name":"Department of Bioengineering, University of Texas at Dallas, Richardson, TX 75080, USA"},{"name":"Department of Biomedical Engineering, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,12,11]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"11","DOI":"10.1016\/0092-8674(92)90115-S","article-title":"Integrins: Versatility, Modulation, and Signaling in Cell Adhesion","volume":"69","author":"Hynes","year":"1992","journal-title":"Cell"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"457","DOI":"10.1038\/s41580-019-0134-2","article-title":"Integrins as Biomechanical Sensors of the Microenvironment","volume":"20","author":"Kechagia","year":"2019","journal-title":"Nat. Rev. Mol. Cell Biol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"14923","DOI":"10.1038\/ncomms14923","article-title":"Anisotropic Forces from Spatially Constrained Focal Adhesions Mediate Contact Guidance Directed Cell Migration","volume":"8","author":"Ray","year":"2017","journal-title":"Nat. Commun."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1607","DOI":"10.1016\/j.cub.2013.06.053","article-title":"Matrix Microarchitecture and Myosin II Determine Adhesion in 3D Matrices","volume":"23","author":"Kubow","year":"2013","journal-title":"Curr. Biol."},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Tang, S.W., Tong, W.Y., Shen, W., Yeung, K.W.K., and Lam, Y.W. (2014). Stringent Requirement for Spatial Arrangement of Extracellular Matrix in Supporting Cell Morphogenesis and Differentiation. BMC Cell Biol., 15.","DOI":"10.1186\/1471-2121-15-10"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"273","DOI":"10.1016\/S0006-3495(93)81364-X","article-title":"Ultrastructure of the Corneal Stroma: A Comparative Study","volume":"64","author":"Meek","year":"1993","journal-title":"Biophys. J."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"83","DOI":"10.1007\/s12551-009-0011-x","article-title":"Corneal Collagen-Its Role in Maintaining Corneal Shape and Transparency","volume":"1","author":"Meek","year":"2009","journal-title":"Biophys. Rev."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"2546","DOI":"10.1016\/j.bpj.2014.10.035","article-title":"3D Collagen Alignment Limits Protrusions to Enhance Breast Cancer Cell Persistence","volume":"107","author":"Riching","year":"2015","journal-title":"Biophys. J."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"33","DOI":"10.1006\/exer.2000.0850","article-title":"The Influence of Corneal Stromal Matrix Proteins on the Migration of Human Corneal Fibroblasts","volume":"71","author":"Andresen","year":"2000","journal-title":"Exp. Eye Res."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"354","DOI":"10.1016\/S0886-3350(02)01609-7","article-title":"Effect of Tenascin and Fibronectin on the Migration of Human Corneal Fibroblasts","volume":"29","author":"Schmidinger","year":"2003","journal-title":"J. Cataract. Refract. Surg."},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Tjahjono, N.S., Subramanian, D., Shihabeddin, T.Z., Hicks, H.D., Varner, V.D., Petroll, W.M., and Schmidtke, D.W. (2025). Effect of Decorin and Aligned Collagen Fibril Topography on TGF-\u03921 Activation of Corneal Keratocytes. Bioengineering, 12.","DOI":"10.3390\/bioengineering12030259"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"664","DOI":"10.1002\/mabi.200900452","article-title":"Response of Human Corneal Fibroblasts on Silk Film Surface Patterns","volume":"10","author":"Gil","year":"2010","journal-title":"Macromol. Biosci."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"748","DOI":"10.1016\/j.jtos.2020.07.020","article-title":"Decellularized Porcine Cornea-Derived Hydrogels for the Regeneration of Epithelium and Stroma in Focal Corneal Defects","volume":"18","author":"Wang","year":"2020","journal-title":"Ocul. Surf."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"369","DOI":"10.1002\/jbm.a.30089","article-title":"Responses of Human Keratocytes to Micro- and Nanostructured Substrates","volume":"71","author":"Teixeira","year":"2004","journal-title":"J. Biomed. Mater. Res. A"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"1373","DOI":"10.1167\/iovs.09-4074","article-title":"Nanoscale Topography-Induced Modulation of Fundamental Cell Behaviors of Rabbit Corneal Keratocytes, Fibroblasts, and Myofibroblasts","volume":"51","author":"Pot","year":"2010","journal-title":"Investig. Ophthalmol. Vis. Sci."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"811","DOI":"10.1167\/iovs.11-7982","article-title":"Substratum Topography Modulates Corneal Fibroblast to Myofibroblast Transformation","volume":"53","author":"Myrna","year":"2012","journal-title":"Investig. Ophthalmol. Vis. Sci."},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Lam, K.H., Shihabeddin, T.Z., Awkal, J.A., Najjar, A.M., Miron-Mendoza, M., Maruri, D.P., Varner, V.D., Petroll, W.M., and Schmidtke, D.W. (2023). Effects of Topography and PDGF on the Response of Corneal Keratocytes to Fibronectin-Coated Surfaces. J. Funct. Biomater., 14.","DOI":"10.3390\/jfb14040217"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"725","DOI":"10.1002\/jbm.a.31519","article-title":"Sub-Micron and Nanoscale Feature Depth Modulates Alignment of Stromal Fibroblasts and Corneal Epithelial Cells in Serum-Rich and Serum-Free Media","volume":"86","author":"Fraser","year":"2008","journal-title":"J. Biomed. Mater. Res. A"},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Zhurenkov, K.E., Lobov, A.A., Bildyug, N.B., Alexander-Sinclair, E.I., Darvish, D.M., Lomert, E.V., Kriger, D.V., Zainullina, B.R., Chabina, A.S., and Khorolskaya, J.I. (2024). Focal Adhesion Maturation Responsible for Behavioral Changes in Human Corneal Stromal Fibroblasts on Fibrillar Substrates. Int. J. Mol. Sci., 25.","DOI":"10.3390\/ijms25168601"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"6679","DOI":"10.1038\/s41598-020-62640-5","article-title":"Influence of Micropatterned Substrates on Keratocyte Phenotype","volume":"10","author":"Bhattacharjee","year":"2020","journal-title":"Sci. Rep."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"110971","DOI":"10.1016\/j.colsurfb.2020.110971","article-title":"Effect of Substrate Topography on the Regulation of Human Corneal Stromal Cells","volume":"190","author":"Bhattacharjee","year":"2020","journal-title":"Colloids Surf. B Biointerfaces"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"4285","DOI":"10.1016\/j.actbio.2012.07.004","article-title":"Focal Adhesion Kinase Knockdown Modulates the Response of Human Corneal Epithelial Cells to Topographic Cues","volume":"8","author":"Dreier","year":"2012","journal-title":"Acta Biomater."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"2551","DOI":"10.1021\/acs.langmuir.3c02676","article-title":"Fabrication of Micropatterns of Aligned Collagen Fibrils","volume":"40","author":"Subramanian","year":"2024","journal-title":"Langmuir"},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Subramanian, D., Tjahjono, N.S., Nammi, S., Miron-Mendoza, M., Varner, V.D., Petroll, W.M., and Schmidtke, D.W. (2025). Effects of Cell Seeding Density, Extracellular Matrix Composition, and Geometry on Yes-Associated Protein Translocation in Corneal Fibroblasts. Int. J. Mol. Sci., 26.","DOI":"10.3390\/ijms26031183"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"293","DOI":"10.4161\/cam.24804","article-title":"Predicting How Cells Spread and Migrate: Focal Adhesion Size Does Matter","volume":"7","author":"Kim","year":"2013","journal-title":"Cell Adhes. Migr."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"1351","DOI":"10.1096\/fj.12-220160","article-title":"Focal Adhesion Size Uniquely Predicts Cell Migration","volume":"27","author":"Kim","year":"2013","journal-title":"FASEB J."},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"Dede Eren, A., Lucassen, A.W.A., Tuvshindorj, U., Truckenm\u00fcller, R., Giselbrecht, S., Eren, E.D., Tas, M.O., Sudarsanam, P., and de Boer, J. (2022). Cells Dynamically Adapt to Surface Geometry by Remodeling Their Focal Adhesions and Actin Cytoskeleton. Front. Cell Dev. Biol., 10.","DOI":"10.3389\/fcell.2022.863721"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"4329","DOI":"10.1091\/mbc.e05-02-0170","article-title":"Cell Adhesion Strengthening: Contributions of Adhesive Area, Integrin Binding, and Focal Adhesion Assembly","volume":"16","author":"Gallant","year":"2005","journal-title":"Mol. Biol. Cell"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"2903","DOI":"10.1016\/j.bpj.2011.11.013","article-title":"Regulation of Cell Adhesion Strength by Peripheral Focal Adhesion Distribution","volume":"101","author":"Elineni","year":"2011","journal-title":"Biophys. J."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"15321","DOI":"10.1038\/ncomms15321","article-title":"YAP Regulates Cell Mechanics by Controlling Focal Adhesion Assembly","volume":"8","author":"Nardone","year":"2017","journal-title":"Nat. Commun."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"3448","DOI":"10.1096\/fj.11-183459","article-title":"Focal Adhesions and Ras Are Functionally and Spatially Integrated to Mediate IL-1 Activation of ERK","volume":"25","author":"Wang","year":"2011","journal-title":"FASEB J."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"565","DOI":"10.1002\/jcp.21237","article-title":"Cell-Matrix Adhesion","volume":"213","author":"Berrier","year":"2006","journal-title":"J. Cell. Physiol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"195","DOI":"10.1002\/jbm.a.32196","article-title":"Interactions with Nanoscale Topography: Adhesion Quantification and Signal Transduction in Cells of Osteogenic and Multipotent Lineage","volume":"91","author":"Biggs","year":"2009","journal-title":"J. Biomed. Mater. Res. A"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"651","DOI":"10.1016\/j.actbio.2013.11.008","article-title":"Osteogenic Lineage Restriction by Osteoprogenitors Cultured on Nanometric Grooved Surfaces: The Role of Focal Adhesion Maturation","volume":"10","author":"Cassidy","year":"2014","journal-title":"Acta Biomater."},{"key":"ref_35","first-page":"2690","article-title":"Identification of Integrins in Cultured Corneal Fibroblasts and in Isolated Keratocytes","volume":"34","author":"Masur","year":"1993","journal-title":"Investig. Ophthalmol. Vis. Sci."},{"key":"ref_36","first-page":"730","article-title":"Corneal Keratocytes: In Situ and in Vitro Organization of Cytoskeletal Contractile Proteins","volume":"35","author":"Jester","year":"1994","journal-title":"Investig. Ophthalmol. Vis. Sci."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"3","DOI":"10.1016\/j.exer.2006.01.010","article-title":"Corneal Integrins and Their Functions","volume":"83","author":"Stepp","year":"2006","journal-title":"Exp. Eye Res."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"035005","DOI":"10.1088\/1748-605X\/ab673c","article-title":"Effects of Aligned Electrospun Fibers with Different Diameters on Hemocompatibility, Cell Behaviors and Inflammation in Vitro","volume":"15","author":"Yu","year":"2020","journal-title":"Biomed. Mater."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"461","DOI":"10.1016\/j.colsurfb.2018.07.045","article-title":"Effects of Aligned and Random Fibers with Different Diameter on Cell Behaviors","volume":"171","author":"Li","year":"2018","journal-title":"Colloids Surf. B Biointerfaces"},{"key":"ref_40","doi-asserted-by":"crossref","unstructured":"Lichtenberg, J.Y., Ramamurthy, E., Young, A.D., Redman, T.P., Leonard, C.E., Das, S.K., Fisher, P.B., Lemmon, C.A., and Hwang, P.Y. (2024). Leader Cells Mechanically Respond to Aligned Collagen Architecture to Direct Collective Migration. PLoS ONE, 19.","DOI":"10.1371\/journal.pone.0296153"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"189","DOI":"10.1016\/j.colsurfb.2011.02.028","article-title":"Nanoscale Topography Reduces Fibroblast Growth, Focal Adhesion Size and Migration-Related Gene Expression on Platinum Surfaces","volume":"85","author":"Pennisi","year":"2011","journal-title":"Colloids Surf. B Biointerfaces"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"9568","DOI":"10.1016\/j.biomaterials.2011.08.077","article-title":"The Effect of Substrate Microtopography on Focal Adhesion Maturation and Actin Organization via the RhoA\/ROCK Pathway","volume":"32","author":"Seo","year":"2011","journal-title":"Biomaterials"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"1881","DOI":"10.1242\/jcs.00383","article-title":"Epithelial Contact Guidance on Well-Defined Micro- and Nanostructured Substrates","volume":"116","author":"Teixeira","year":"2003","journal-title":"J. Cell Sci."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"1791","DOI":"10.1021\/acsomega.8b03608","article-title":"Cell Type-Dependent Integrin Distribution in Adhesion and Migration Responses on Protein-Coated Microgrooved Substrates","volume":"4","author":"Sales","year":"2019","journal-title":"ACS Omega"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"99","DOI":"10.1007\/s10544-019-0436-3","article-title":"A High-Throughput Microfluidic Method for Fabricating Aligned Collagen Fibrils to Study Keratocyte Behavior","volume":"21","author":"Lam","year":"2019","journal-title":"Biomed. Microdevices"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"1850","DOI":"10.1167\/iovs.02-0973","article-title":"Myofibroblast Differentiation of Normal Human Keratocytes and HTERT, Extended-Life","volume":"44","author":"Jester","year":"2003","journal-title":"Investig. Ophthalmol. Vis. Sci."},{"key":"ref_47","doi-asserted-by":"crossref","unstructured":"Maruri, D.P., Iyer, K.S., Schmidtke, D.W., Petroll, W.M., and Varner, V.D. (2022). Signaling Downstream of Focal Adhesions Regulates Stiffness-Dependent Differences in the TGF-\u03921-Mediated Myofibroblast Differentiation of Corneal Keratocytes. Front. Cell Dev. Biol., 10.","DOI":"10.3389\/fcell.2022.886759"},{"key":"ref_48","unstructured":"Subramanian, D. (2024). Investigating the Role of Extracellular Matrix Topography, Geometry and Composition of Corneal Fibroblast Behavior. [Ph.D. Thesis, University of Texas at Dallas]."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/26\/24\/11935\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,12,13]],"date-time":"2025-12-13T05:20:15Z","timestamp":1765603215000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/26\/24\/11935"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,12,11]]},"references-count":48,"journal-issue":{"issue":"24","published-online":{"date-parts":[[2025,12]]}},"alternative-id":["ijms262411935"],"URL":"https:\/\/doi.org\/10.3390\/ijms262411935","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,12,11]]}}}