{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,9]],"date-time":"2026-06-09T23:33:59Z","timestamp":1781048039033,"version":"3.54.1"},"reference-count":43,"publisher":"MDPI AG","issue":"9","license":[{"start":{"date-parts":[[2025,9,12]],"date-time":"2025-09-12T00:00:00Z","timestamp":1757635200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Marie Sk\u0142odowska Curie International Training Network (ITN)","award":["#955735"],"award-info":[{"award-number":["#955735"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Gels"],"abstract":"<jats:p>Little is known about cartilaginous endplate (CEP) mechanobiology or how it changes in a catabolic microenvironment, partly due to difficulties in conducting mechanotransduction in vitro. Recent studies have found blended collagen\u2013agarose hydrogels to offer improved mechanotransduction in chondrocytes compared to agarose alone. It was hypothesized that blended collagen\u2013agarose hydrogels would be sufficient to improve the mechanobiological response in CEP cells relative to that in agarose alone, while maintaining the chondrocyte phenotype and ability to respond to pro-inflammatory stimulation. Thus, human CEP cells were seeded into blended 2% agarose and 2 mg\/mL type I collagen hydrogels, followed by culture with dynamic compression up to 7% and stimulation with TNF. Results confirmed CEP cells retained a rounded phenotype and high cell viability during culture in blended collagen\u2013agarose hydrogels. Additionally, TNF induced a catabolic response through downregulation of pericellular marker COL6A1 and anabolic markers ACAN and COL2A1. No significant changes were seen due to dynamic compression, suggesting addition of collagen to agarose was not sufficient to induce mechanotransduction in human CEP cells in this study. However, blended collagen\u2013agarose hydrogels increased stiffness by 4\u00d7 and gene expression of key cartilage marker SOX9 and physioosmotic mechanosensor TRPV4, offering an improvement on agarose alone.<\/jats:p>","DOI":"10.3390\/gels11090736","type":"journal-article","created":{"date-parts":[[2025,9,15]],"date-time":"2025-09-15T11:51:43Z","timestamp":1757937103000},"page":"736","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Exploring Mechanotransduction and Inflammation in Human Cartilaginous Endplate Cells in Blended Collagen\u2013Agarose Hydrogels Under Cyclic Compression"],"prefix":"10.3390","volume":"11","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-5328-667X","authenticated-orcid":false,"given":"Katherine B.","family":"Crump","sequence":"first","affiliation":[{"name":"Tissue Engineering for Orthopaedics & Mechanobiology, Bone & Joint Program, Department for BioMedical Research, Faculty of Medicine, University of Bern, 3008 Bern, Switzerland"},{"name":"Graduate School for Cellular and Biomedical Sciences, University of Bern, 3012 Bern, Switzerland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0003-8409-4613","authenticated-orcid":false,"given":"Chlo\u00e9","family":"Chapallaz","sequence":"additional","affiliation":[{"name":"Tissue Engineering for Orthopaedics & Mechanobiology, Bone & Joint Program, Department for BioMedical Research, Faculty of Medicine, University of Bern, 3008 Bern, Switzerland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9998-3275","authenticated-orcid":false,"given":"Ahmad","family":"Alminnawi","sequence":"additional","affiliation":[{"name":"GIGA In Silico Medicine, University of Li\u00e8ge, 4000 Li\u00e8ge, Belgium"},{"name":"Skeletal Biology and Engineering Research Center, KU Leuven, 3000 Leuven, Belgium"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6858-8213","authenticated-orcid":false,"given":"Paola","family":"Bermudez-Lekerika","sequence":"additional","affiliation":[{"name":"Tissue Engineering for Orthopaedics & Mechanobiology, Bone & Joint Program, Department for BioMedical Research, Faculty of Medicine, University of Bern, 3008 Bern, Switzerland"},{"name":"Graduate School for Cellular and Biomedical Sciences, University of Bern, 3012 Bern, Switzerland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8180-1445","authenticated-orcid":false,"given":"Liesbet","family":"Geris","sequence":"additional","affiliation":[{"name":"GIGA In Silico Medicine, University of Li\u00e8ge, 4000 Li\u00e8ge, Belgium"},{"name":"Skeletal Biology and Engineering Research Center, KU Leuven, 3000 Leuven, Belgium"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3446-7621","authenticated-orcid":false,"given":"J\u00e9r\u00f4me","family":"Noailly","sequence":"additional","affiliation":[{"name":"BCN MedTech, Department of Engineering, Universitat Pompeu Fabra, 08002 Barcelona, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9005-0655","authenticated-orcid":false,"given":"Benjamin","family":"Gantenbein","sequence":"additional","affiliation":[{"name":"Tissue Engineering for Orthopaedics & Mechanobiology, Bone & Joint Program, Department for BioMedical Research, Faculty of Medicine, University of Bern, 3008 Bern, Switzerland"},{"name":"Department of Orthopedic Surgery & Traumatology, Faculty of Medicine, Inselspital, Bern University Hospital, University of Bern, 3010 Bern, Switzerland"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,9,12]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Bermudez-Lekerika, P., Crump, K.B., Tseranidou, S., N\u00fcesch, A., Kanelis, E., Alminnawi, A., Baumgartner, L., Mu\u00f1oz-Moya, E., Compte, R., and Gualdi, F. (2022). Immuno-Modulatory Effects of Intervertebral Disc Cells. Front. Cell Dev. Biol., 10.","DOI":"10.3389\/fcell.2022.924692"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1796","DOI":"10.1007\/s00586-011-1827-1","article-title":"The Effects of Dynamic Loading on the Intervertebral Disc","volume":"20","author":"Chan","year":"2011","journal-title":"Eur. Spine J."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"E78","DOI":"10.1097\/BRS.0000000000001699","article-title":"Influence of Complex Loading Conditions on Intervertebral Disc Failure","volume":"42","author":"Casaroli","year":"2017","journal-title":"Spine"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1363","DOI":"10.1007\/s00586-016-4416-5","article-title":"A New Dynamic Six Degrees of Freedom Disc-Loading Simulator Allows to Provoke Disc Damage and Herniation","volume":"25","author":"Wilke","year":"2016","journal-title":"Eur. Spine J."},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Baumgartner, L., Wuertz-Kozak, K., Le Maitre, C.L., Wignall, F., Richardson, S.M., Hoyland, J., Ruiz Wills, C., Gonz\u00e1lez Ballester, M.A., Neidlin, M., and Alexopoulos, L.G. (2021). Multiscale Regulation of the Intervertebral Disc: Achievements in Experimental, In Silico, and Regenerative Research. Int. J. Mol. Sci., 22.","DOI":"10.3390\/ijms22020703"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"2351","DOI":"10.1111\/jcmm.12377","article-title":"Mechanotransduction in Intervertebral Discs","volume":"18","author":"Tsai","year":"2014","journal-title":"J. Cell Mol. Med."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"460","DOI":"10.1002\/art.24248","article-title":"Altered Integrin Mechanotransduction in Human Nucleus Pulposus Cells Derived from Degenerated Discs","volume":"60","author":"Frain","year":"2009","journal-title":"Arthritis Rheum."},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Gilbert, H.T.J., Nagra, N.S., Freemont, A.J., Millward-Sadler, S.J., and Hoyland, J.A. (2013). Integrin\u2013Dependent Mechanotransduction in Mechanically Stimulated Human Annulus Fibrosus Cells: Evidence for an Alternative Mechanotransduction Pathway Operating with Degeneration. PLoS ONE, 8.","DOI":"10.1371\/journal.pone.0072994"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"eaba2368","DOI":"10.1126\/sciadv.aba2368","article-title":"Actomyosin Contractility Confers Mechanoprotection Against TNF\u03b1-Induced Disruption of the Intervertebral Disc","volume":"6","author":"Hernandez","year":"2020","journal-title":"Sci. Adv."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"R8","DOI":"10.1186\/ar3229","article-title":"The Involvement of Interleukin-1 and Interleukin-4 in the Response of Human Annulus Fibrosus Cells to Cyclic Tensile Strain: An Altered Mechanotransduction Pathway with Degeneration","volume":"13","author":"Gilbert","year":"2011","journal-title":"Arthritis Res. Ther."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"550","DOI":"10.1016\/j.jbiomech.2016.01.007","article-title":"Human Cartilage Endplate Permeability Varies with Degeneration and Intervertebral Disc Site","volume":"49","author":"DeLucca","year":"2016","journal-title":"J. Biomech."},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Wu, Y., Cisewski, S.E., Sachs, B.L., Pellegrini, V.D., Kern, M.J., Slate, E.H., and Yao, H. (2015). The Region-Dependent Biomechanical and Biochemical Properties of Bovine Cartilaginous Endplate. J. Biomech., 48.","DOI":"10.1016\/j.jbiomech.2015.07.005"},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Wu, Y., Glaser, J., and Yao, H. (2008, January 25\u201329). Effects of Endplate and Mechanical Loading on Solute Transport in Intervertebral Disc. Proceedings of the ASME Summer Bioengineering Conference, SBC2008, Marco Island, FL, USA.","DOI":"10.1115\/SBC2008-193111"},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Ruiz Wills, C., Foata, B., Gonz\u00e1lez Ballester, M.\u00c1., Karppinen, J., and Noailly, J. (2018). Theoretical Explorations Generate New Hypotheses About the Role of the Cartilage Endplate in Early Intervertebral Disk Degeneration. Front. Physiol., 9.","DOI":"10.3389\/fphys.2018.01210"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"1362","DOI":"10.1007\/s00586-017-4955-4","article-title":"ISSLS Prize in Basic Science 2017: Intervertebral Disc\/Bone Marrow Cross-Talk with Modic Changes","volume":"26","author":"Dudli","year":"2017","journal-title":"Eur. Spine J."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"92","DOI":"10.1007\/s005860050217","article-title":"The Vertebral End-Plate: What Do We Know?","volume":"9","author":"Moore","year":"2000","journal-title":"Eur. Spine J."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"166","DOI":"10.1097\/00007632-198902000-00005","article-title":"Biochemical and Structural Properties of the Cartilage End-Plate and Its Relation to the Intervertebral Disc","volume":"14","author":"Roberts","year":"1989","journal-title":"Spine"},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Crump, K.B., Kanelis, E., Segarra-Queralt, M., Pascuet-Fontanet, A., Bermudez-Lekerika, P., Alminnawi, A., Geris, L., Alexopoulos, L.G., Noailly, J., and Gantenbein, B. (2025). TNF Induces Catabolism in Human Cartilaginous Endplate Cells in 3D Agarose Culture Under Dynamic Compression. Sci. Rep., 15.","DOI":"10.1038\/s41598-025-00538-w"},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Cambria, E., Brunner, S., Heusser, S., Fisch, P., Hitzl, W., Ferguson, S.J., and Wuertz-Kozak, K. (2020). Cell-Laden Agarose-Collagen Composite Hydrogels for Mechanotransduction Studies. Front. Bioeng. Biotechnol., 8.","DOI":"10.3389\/fbioe.2020.00346"},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Bougault, C., Paumier, A., Aubert-Foucher, E., and Mallein-Gerin, F. (2008). Molecular Analysis of Chondrocytes Cultured in Agarose in Response to Dynamic Compression. BMC Biotechnol., 8.","DOI":"10.1186\/1472-6750-8-71"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"252","DOI":"10.1115\/1.429656","article-title":"Functional Tissue Engineering of Articular Cartilage Through Dynamic Loading of Chondrocyte-Seeded Agarose Gels","volume":"122","author":"Mauck","year":"2000","journal-title":"J. Biomech. Eng."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"928","DOI":"10.1038\/nprot.2009.63","article-title":"Investigating Conversion of Mechanical Force into Biochemical Signaling in Three-Dimensional Chondrocyte Cultures","volume":"4","author":"Bougault","year":"2009","journal-title":"Nat. Protoc."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"1875","DOI":"10.1016\/j.biomaterials.2009.10.047","article-title":"Probing Cellular Mechanobiology in Three-Dimensional Culture with Collagen\u2013Agarose Matrices","volume":"31","author":"Ulrich","year":"2010","journal-title":"Biomaterials"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"4118","DOI":"10.1016\/j.biomaterials.2012.02.044","article-title":"Laminin Active Peptide\/Agarose Matrices as Multifunctional Biomaterials for Tissue Engineering","volume":"33","author":"Yamada","year":"2012","journal-title":"Biomaterials"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"1152","DOI":"10.1016\/j.ijbiomac.2018.09.065","article-title":"Hydrogels of Agarose, and Methacrylated Gelatin and Hyaluronic Acid Are More Supportive for In Vitro Meniscus Regeneration than Three Dimensional Printed Polycaprolactone Scaffolds","volume":"122","author":"Bahcecioglu","year":"2019","journal-title":"Int. J. Biol. Macromol."},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Torabi Rahvar, P., Abdekhodaie, M.J., Jooybar, E., and Gantenbein, B. (2024). An Enzymatically Crosslinked Collagen Type II\/Hyaluronic Acid Hybrid Hydrogel: A Biomimetic Cell Delivery System for Cartilage Tissue Engineering. Int. J. Biol. Macromol., 279.","DOI":"10.1016\/j.ijbiomac.2024.134614"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"e2001847","DOI":"10.1002\/adhm.202001847","article-title":"Development of a Biomimetic Hydrogel Based on Predifferentiated Mesenchymal Stem-Cell-Derived ECM for Cartilage Tissue Engineering","volume":"10","author":"Antich","year":"2021","journal-title":"Adv. Healthc. Mater."},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Jacobs, C.A.M., Kamali, S.A., Abdelgawad, A.M., Meij, B.P., Ghazanfari, S., Tryfonidou, M.A., Jockenhoevel, S., and Ito, K. (2023). Mechanical Characterization of a Novel Biomimetic Artificial Disc for the Cervical Spine. J. Mech. Behav. Biomed. Mater., 142.","DOI":"10.1016\/j.jmbbm.2023.105808"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"E260","DOI":"10.1097\/BRS.0000000000001791","article-title":"Regenerative Intervertebral Disc Endplate Based on Biomimetic Three-Dimensional Scaffolds","volume":"42","author":"Yuan","year":"2017","journal-title":"Spine"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"1132","DOI":"10.1007\/s10439-014-1183-5","article-title":"Encapsulation of Chondrocytes in High-Stiffness Agarose Microenvironments for In Vitro Modeling of Osteoarthritis Mechanotransduction","volume":"43","author":"Jutila","year":"2015","journal-title":"Ann. Biomed. Eng."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"670","DOI":"10.1159\/000514985","article-title":"A High-Throughput Mechanical Activator for Cartilage Engineering Enables Rapid Screening of in Vitro Response of Tissue Models to Physiological and Supra-Physiological Loads","volume":"211","author":"Capuana","year":"2022","journal-title":"Cells Tissues Organs"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"451","DOI":"10.1007\/s11926-014-0451-6","article-title":"The Mechanobiology of Articular Cartilage: Bearing the Burden of Osteoarthritis","volume":"16","author":"Leddy","year":"2014","journal-title":"Curr. Rheumatol. Rep."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"1316","DOI":"10.1073\/pnas.1319569111","article-title":"TRPV4-Mediated Mechanotransduction Regulates the Metabolic Response of Chondrocytes to Dynamic Loading","volume":"111","author":"Leddy","year":"2014","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"32158","DOI":"10.1074\/jbc.M706158200","article-title":"Functional Gene Screening System Identified TRPV4 as a Regulator of Chondrogenic Differentiation","volume":"282","author":"Muramatsu","year":"2007","journal-title":"J. Biol. Chem."},{"key":"ref_35","doi-asserted-by":"crossref","unstructured":"Anderson, D.E., and Johnstone, B. (2017). Dynamic Mechanical Compression of Chondrocytes for Tissue Engineering: A Critical Review. Front. Bioeng. Biotechnol., 5.","DOI":"10.3389\/fbioe.2017.00076"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"1207","DOI":"10.1016\/j.joca.2007.03.023","article-title":"Structure of Pericellular Matrix Around Agarose-Embedded Chondrocytes","volume":"15","author":"DiMicco","year":"2007","journal-title":"Osteoarthr. Cartil."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"179","DOI":"10.1016\/j.joca.2005.09.002","article-title":"Chondrogenic Differentiation and Functional Maturation of Bovine Mesenchymal Stem Cells in Long-Term Agarose Culture","volume":"14","author":"Mauck","year":"2006","journal-title":"Osteoarthr. Cartil."},{"key":"ref_38","doi-asserted-by":"crossref","unstructured":"Tasci, A., Ettinger, L., Ferguson, S., and B\u00fcchler, P. (2009, January 17\u201321). The Role of Agarose Mechanical Response on the Matrix Synthesis of Nucleus Pulposus Cells: A Pilot Study. Proceedings of the ASME 2009 Summer Bioengineering Conference, Parts A and B, Olympic Valley, CA, USA.","DOI":"10.1115\/SBC2009-206763"},{"key":"ref_39","doi-asserted-by":"crossref","unstructured":"Frauchiger, D., May, R., Bakirci, E., Tekari, A., Chan, S., W\u00f6ltje, M., Benneker, L., and Gantenbein, B. (2018). Genipin-Enhanced Fibrin Hydrogel and Novel Silk for Intervertebral Disc Repair in a Loaded Bovine Organ Culture Model. J. Funct. Biomater., 9.","DOI":"10.3390\/jfb9030040"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"676","DOI":"10.1038\/nmeth.2019","article-title":"Fiji: An Open-Source Platform for Biological-Image Analysis","volume":"9","author":"Schindelin","year":"2012","journal-title":"Nat. Methods"},{"key":"ref_41","doi-asserted-by":"crossref","unstructured":"Croft, A.S., Roth, Y., Oswald, K.A.C., \u0106orluka, S., Bermudez-Lekerika, P., and Gantenbein, B. (2021). In Situ Cell Signalling of the Hippo-YAP\/TAZ Pathway in Reaction to Complex Dynamic Loading in an Intervertebral Disc Organ Culture. Int. J. Mol. Sci., 22.","DOI":"10.3390\/ijms222413641"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"210","DOI":"10.1002\/term.307","article-title":"Numerical Assessment on the Effective Mechanical Stimuli for Matrix-Associated Metabolism in Chondrocyte-Seeded Constructs","volume":"5","author":"Tasci","year":"2011","journal-title":"J. Tissue Eng. Regen. Med."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"593","DOI":"10.1016\/S0021-9290(02)00437-2","article-title":"New Insight into Deformation-Dependent Hydraulic Permeability of Gels and Cartilage, and Dynamic Behavior of Agarose Gels in Confined Compression","volume":"36","author":"Gu","year":"2003","journal-title":"J. Biomech."}],"container-title":["Gels"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2310-2861\/11\/9\/736\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T18:44:58Z","timestamp":1760035498000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2310-2861\/11\/9\/736"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,9,12]]},"references-count":43,"journal-issue":{"issue":"9","published-online":{"date-parts":[[2025,9]]}},"alternative-id":["gels11090736"],"URL":"https:\/\/doi.org\/10.3390\/gels11090736","relation":{},"ISSN":["2310-2861"],"issn-type":[{"value":"2310-2861","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,9,12]]}}}