{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,19]],"date-time":"2026-09-19T10:41:25Z","timestamp":1789814485984,"version":"4.0.1"},"reference-count":75,"publisher":"MDPI AG","issue":"4","license":[{"start":{"date-parts":[[2025,4,2]],"date-time":"2025-04-02T00:00:00Z","timestamp":1743552000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Gels"],"abstract":"<jats:p>Glioblastoma (GBM) is a highly aggressive and malignant brain tumor, characterized by hypoxia in its microenvironment, which drives its growth and resistance to treatments. Hypoxia-inducible factor 1 (HIF-1) plays a central role in GBM progression by regulating cellular adaptation to low oxygen availability, promoting processes such as angiogenesis and cell invasion. However, studying and modeling GBM under hypoxic conditions is complex, especially due to the limitations of animal models. In this study, we developed a glioma spheroid model using an alginate\u2013gelatin hydrogel scaffold, which enabled the simulation of hypoxic conditions within the tumor. The scaffold-based model demonstrated high reproducibility, facilitating the analysis of HIF-1\u03b1 expression, a key protein in the hypoxic response of GBM. Furthermore, cell viability, the microstructural features of the encapsulated spheroids, and the water absorption rate of the hydrogel were assessed. Our findings validate the three-dimensional (3D) glioblastoma spheroids model as a valuable platform for studying hypoxia in GBM and evaluating new therapies. This approach could offer a more accessible and specific alternative for studying the tumor microenvironment and therapeutic resistance in GBM.<\/jats:p>","DOI":"10.3390\/gels11040263","type":"journal-article","created":{"date-parts":[[2025,4,2]],"date-time":"2025-04-02T07:47:44Z","timestamp":1743580064000},"page":"263","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["Alginate\u2013Gelatin Hydrogel Scaffold Model for Hypoxia Induction in Glioblastoma Embedded Spheroids"],"prefix":"10.3390","volume":"11","author":[{"given":"Janette","family":"del Roc\u00edo Aguilera-Marquez","sequence":"first","affiliation":[{"name":"Unidad de Biotecnolog\u00eda M\u00e9dica y Farmac\u00e9utica, Centro de Investigaci\u00f3n y Asistencia en Tecnolog\u00eda y Dise\u00f1o del Estado de Jalisco (CIATEJ), Guadalajara 44270, Mexico"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alejandro","family":"Manzanares-Guzm\u00e1n","sequence":"additional","affiliation":[{"name":"Unidad de Biotecnolog\u00eda M\u00e9dica y Farmac\u00e9utica, Centro de Investigaci\u00f3n y Asistencia en Tecnolog\u00eda y Dise\u00f1o del Estado de Jalisco (CIATEJ), Guadalajara 44270, Mexico"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3018-6013","authenticated-orcid":false,"given":"Lorena","family":"Garc\u00eda-Uriostegui","sequence":"additional","affiliation":[{"name":"SECIHTI-Secretar\u00eda de Ciencia, Humanidades, Tecnolog\u00eda e Innovaci\u00f3n-Centro Universitario de Ciencias Exactas e Ingenier\u00edas, Universidad de Guadalajara, Guadalajara 44430, Mexico"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0918-788X","authenticated-orcid":false,"given":"Alejandro A.","family":"Canales-Aguirre","sequence":"additional","affiliation":[{"name":"Unidad de Biotecnolog\u00eda M\u00e9dica y Farmac\u00e9utica, Centro de Investigaci\u00f3n y Asistencia en Tecnolog\u00eda y Dise\u00f1o del Estado de Jalisco (CIATEJ), Guadalajara 44270, Mexico"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0540-0287","authenticated-orcid":false,"given":"Tanya A.","family":"Camacho-Villegas","sequence":"additional","affiliation":[{"name":"Unidad de Biotecnolog\u00eda M\u00e9dica y Farmac\u00e9utica, Centro de Investigaci\u00f3n y Asistencia en Tecnolog\u00eda y Dise\u00f1o del Estado de Jalisco (CIATEJ), Guadalajara 44270, Mexico"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8872-8335","authenticated-orcid":false,"given":"Pavel H.","family":"Lugo-Fabres","sequence":"additional","affiliation":[{"name":"SECIHTI-Secretar\u00eda de Ciencia, Humanidades, Tecnolog\u00eda e Innovaci\u00f3n-Unidad de Biotecnolog\u00eda M\u00e9dica y Farmac\u00e9utica, Centro de Investigaci\u00f3n y Asistencia en Tecnolog\u00eda y Dise\u00f1o del Estado de Jalisco (CIATEJ), Guadalajara 44270, Mexico"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,4,2]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"8596","DOI":"10.1021\/acs.jmedchem.1c01946","article-title":"Glioblastoma: Current status, emerging targets, and recent advances","volume":"65","author":"Thakur","year":"2022","journal-title":"J. Med. Chem."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.canlet.2015.03.015","article-title":"Mechanisms regulating glioma invasion","volume":"362","author":"Paw","year":"2015","journal-title":"Cancer Lett."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"142","DOI":"10.1186\/s13046-022-02349-7","article-title":"Emerging therapies for glioblastoma: Current state and future directions","volume":"41","author":"Rong","year":"2022","journal-title":"J. Exp. Clin. Cancer Res."},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Dom\u00e8nech, M., Hern\u00e1ndez, A., Plaja, A., Mart\u00ednez-Balibrea, E., and Bala\u00f1\u00e0, C. (2021). Hypoxia: The Cornerstone of Glioblastoma. Int. J. Mol. Sci., 22.","DOI":"10.3390\/ijms222212608"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"134","DOI":"10.1215\/S1152851704001115","article-title":"Hypoxia and the hypoxia-inducible-factor pathway in glioma growth and angiogenesis","volume":"7","author":"Kaur","year":"2005","journal-title":"Neuro Oncol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1562","DOI":"10.1038\/s41416-020-1021-5","article-title":"Evidence for hypoxia increasing the tempo of evolution in glioblastoma","volume":"123","author":"Grimes","year":"2020","journal-title":"Br. J. Cancer"},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Cancer Genome Atlas Research Network (2011). Integrated genomic analyses of ovarian carcinoma. Nature, 474, 609\u2013615.","DOI":"10.1038\/nature10166"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"993","DOI":"10.1016\/S0006-2952(02)01168-1","article-title":"Signal transduction to hypoxia-inducible factor 1","volume":"64","author":"Semenza","year":"2002","journal-title":"Biochem. Pharmacol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1260","DOI":"10.3892\/ijo.2012.1555","article-title":"Differential expression of HIF-1 in glioblastoma multiforme and anaplastic astrocytoma","volume":"41","author":"Mayer","year":"2012","journal-title":"Int. J. Oncol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"176","DOI":"10.1152\/physiol.00001.2004","article-title":"Hydroxylation of HIF-1: Oxygen sensing at the molecular level","volume":"19","author":"Semenza","year":"2004","journal-title":"Physiology"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1038\/emm.2004.1","article-title":"Hypoxia-inducible factor (HIF-1)alpha: Its protein stability and biological functions","volume":"36","author":"Lee","year":"2004","journal-title":"Exp. Mol. Med."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"321","DOI":"10.1093\/jnen\/61.4.321","article-title":"Impact of genotype and morphology on the prognosis of glioblastoma","volume":"61","author":"Schmidt","year":"2002","journal-title":"J. Neuropathol. Exp. Neurol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"210","DOI":"10.1046\/j.1365-2990.2002.00391.x","article-title":"Expression of hypoxia-inducible factor 1alpha in tumours of patients with glioblastoma","volume":"28","author":"Hilton","year":"2002","journal-title":"Neuropathol. Appl. Neurobiol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"398","DOI":"10.1016\/j.radonc.2007.05.003","article-title":"Expression patterns of the hypoxia-related genes osteopontin, CA9, erythropoietin, VEGF and HIF-1alpha in human glioma in vitro and in vivo","volume":"83","author":"Said","year":"2007","journal-title":"Radiother. Oncol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"e412","DOI":"10.1038\/cddis.2012.153","article-title":"BMP2 sensitizes glioblastoma stem-like cells to Temozolomide by affecting HIF-1\u03b1 stability and MGMT expression","volume":"3","author":"Persano","year":"2012","journal-title":"Cell Death Dis."},{"key":"ref_16","first-page":"22073","article-title":"Clinical and prognostic significance of HIF-1\u03b1 in glioma patients: A meta-analysis","volume":"8","author":"Liu","year":"2015","journal-title":"Int. J. Clin. Exp. Med."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"3379","DOI":"10.18632\/oncotarget.6582","article-title":"Monoamine oxidase B levels are highly expressed in human gliomas and are correlated with the expression of HiF-1\u03b1 and with transcription factors Sp1 and Sp3","volume":"7","author":"Sharpe","year":"2016","journal-title":"Oncotarget"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"1728","DOI":"10.1093\/carcin\/bgt125","article-title":"GRIM-19 opposes reprogramming of glioblastoma cell metabolism via HIF1\u03b1 destabilization","volume":"34","author":"Liu","year":"2013","journal-title":"Carcinogenesis"},{"key":"ref_19","first-page":"1759","article-title":"The HIF-1\u03b1\/miR-224-3p\/ATG5 axis affects cell mobility and chemosensitivity by regulating hypoxia-induced protective autophagy in glioblastoma and astrocytoma","volume":"41","author":"Huang","year":"2019","journal-title":"Oncol. Rep."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"992","DOI":"10.1038\/s41419-020-03150-0","article-title":"The HIF1\u03b1\/HIF2\u03b1-miR210-3p network regulates glioblastoma cell proliferation, dedifferentiation and chemoresistance through EGF under hypoxic conditions","volume":"11","author":"Wang","year":"2020","journal-title":"Cell Death Dis."},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Zhang, J., Zhang, Y., Mo, F., Patel, G., Butterworth, K., Shao, C., and Prise, K.M. (2021). The Roles of HIF-1\u03b1 in Radiosensitivity and Radiation-Induced Bystander Effects Under Hypoxia. Front. Cell Dev. Biol., 9.","DOI":"10.3389\/fcell.2021.637454"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"148","DOI":"10.1016\/j.yexcr.2016.04.011","article-title":"Downregulation of HIF-1a sensitizes U251 glioma cells to the temozolomide (TMZ) treatment","volume":"343","author":"Tang","year":"2016","journal-title":"Exp. Cell Res."},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"B\u00e9dard, P., Gauvin, S., Ferland, K., Caneparo, C., Pellerin, \u00c8., Chabaud, S., and Bolduc, S. (2020). Innovative Human Three-Dimensional Tissue-Engineered Models as an Alternative to Animal Testing. Bioengineering, 7.","DOI":"10.3390\/bioengineering7030115"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"103426","DOI":"10.1016\/j.drudis.2022.103426","article-title":"Advances in 3D culture systems for therapeutic discovery and development in brain cancer","volume":"28","author":"Wanigasekara","year":"2023","journal-title":"Drug Discov. Today"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"e2100157","DOI":"10.1002\/ppap.202100157","article-title":"Plasma induced reactive oxygen species-dependent cytotoxicity in glioblastoma 3D tumourspheres","volume":"19","author":"Wanigasekara","year":"2022","journal-title":"Plasma Processes Polym."},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Chhetri, A., Rispoli, J.V., and Leli\u00e8vre, S.A. (2021). 3D Cell Culture for the Study of Microenvironment-Mediated Mechanostimuli to the Cell Nucleus: An Important Step for Cancer Research. Front. Mol. Biosci., 8.","DOI":"10.3389\/fmolb.2021.628386"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1033","DOI":"10.1016\/j.trecan.2021.06.009","article-title":"Tumour microenvironment 3D-modelling: Simplicity to complexity and back again","volume":"7","author":"Carter","year":"2021","journal-title":"Trends Cancer"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"117784","DOI":"10.1016\/j.lfs.2020.117784","article-title":"Methodological aspects and pharmacological applications of three-dimensional cancer cell cultures and organoids","volume":"254","author":"Foglietta","year":"2020","journal-title":"Life Sci."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"1367","DOI":"10.1093\/neuonc\/now045","article-title":"Large-scale assessment of the gliomasphere model system","volume":"18","author":"Laks","year":"2016","journal-title":"Neuro Oncol."},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"Jensen, C., and Teng, Y. (2020). Is It Time to Start Transitioning From 2D to 3D Cell Culture?. Front. Mol. Biosci., 7.","DOI":"10.3389\/fmolb.2020.00033"},{"key":"ref_31","doi-asserted-by":"crossref","unstructured":"Colombo, E., and Cattaneo, M.G. (2021). Multicellular 3D Models to Study Tumour-Stroma Interactions. Int. J. Mol. Sci., 22.","DOI":"10.3390\/ijms22041633"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"5074","DOI":"10.7150\/thno.42441","article-title":"3D approaches to model the tumor microenvironment of pancreatic cancer","volume":"10","author":"Kieler","year":"2020","journal-title":"Theranostics"},{"key":"ref_33","doi-asserted-by":"crossref","unstructured":"Alzeeb, G., Metges, J.P., Corcos, L., and Le Jossic-Corcos, C. (2020). Three-Dimensional Culture Systems in Gastric Cancer Research. Cancers, 12.","DOI":"10.3390\/cancers12102800"},{"key":"ref_34","first-page":"6999","article-title":"Three-dimensional cell culture: A powerful tool in tumor research and drug discovery","volume":"14","author":"Lv","year":"2017","journal-title":"Oncol. Lett."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"2262","DOI":"10.1002\/bit.27356","article-title":"Three-dimensional culture models to study drug resistance in breast cancer","volume":"117","author":"Fisher","year":"2020","journal-title":"Biotechnol. Bioeng."},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"Ruiz-Garcia, H., Alvarado-Estrada, K., Schiapparelli, P., Quinones-Hinojosa, A., and Trifiletti, D.M. (2020). Engineering Three-Dimensional Tumor Models to Study Glioma Cancer Stem Cells and Tumor Microenvironment. Front. Cell Neurosci., 14.","DOI":"10.3389\/fncel.2020.558381"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"1972","DOI":"10.1016\/j.cell.2018.11.021","article-title":"Organoid Modeling of the Tumor Immune Microenvironment","volume":"175","author":"Neal","year":"2018","journal-title":"Cell"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"400","DOI":"10.1039\/C9NH00628A","article-title":"3D cultures for modeling nanomaterial-based photothermal therapy","volume":"5","author":"Darrigues","year":"2020","journal-title":"Nanoscale Horiz."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"100753","DOI":"10.1016\/j.drup.2021.100753","article-title":"In vitro biomimetic models for glioblastoma-a promising tool for drug response studies","volume":"55","author":"Dragoj","year":"2021","journal-title":"Drug Resist. Updat."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"1336","DOI":"10.1038\/ncb3429","article-title":"Tissue mechanics promote IDH1-dependent HIF1\u03b1-tenascin C feedback to regulate glioblastoma aggression","volume":"18","author":"Miroshnikova","year":"2016","journal-title":"Nat. Cell Biol."},{"key":"ref_41","doi-asserted-by":"crossref","unstructured":"Klein, E., Hau, A.C., Oudin, A., Golebiewska, A., and Niclou, S.P. (2020). Glioblastoma Organoids: Pre-Clinical Applications and Challenges in the Context of Immunotherapy. Front. Oncol., 10.","DOI":"10.3389\/fonc.2020.604121"},{"key":"ref_42","doi-asserted-by":"crossref","unstructured":"Paolillo, M., Comincini, S., and Schinelli, S. (2021). In Vitro Glioblastoma Models: A Journey into the Third Dimension. Cancers, 13.","DOI":"10.3390\/cancers13102449"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"3173","DOI":"10.1002\/adhm.201600684","article-title":"Culture on 3D Chitosan-Hyaluronic Acid Scaffolds Enhances Stem Cell Marker Expression and Drug Resistance in Human Glioblastoma Cancer Stem Cells","volume":"5","author":"Wang","year":"2016","journal-title":"Adv. Healthc. Mater."},{"key":"ref_44","doi-asserted-by":"crossref","unstructured":"Bahraminasab, M., Asgharzade, S., Doostmohamadi, A., Satari, A., Hasannejad, F., and Arab, S. (2024). Development of a hydrogel-based three-dimensional (3D) glioblastoma cell lines culture as a model system for CD73 inhibitor response study. Biomed. Eng. Online, 23.","DOI":"10.1186\/s12938-024-01320-1"},{"key":"ref_45","doi-asserted-by":"crossref","unstructured":"\u0141abowska, M.B., Cierluk, K., Jankowska, A.M., Kulbacka, J., Detyna, J., and Michalak, I. (2021). A Review on the Adaption of Alginate-Gelatin Hydrogels for 3D Cultures and Bioprinting. Materials, 14.","DOI":"10.3390\/ma14040858"},{"key":"ref_46","doi-asserted-by":"crossref","unstructured":"Law, A.M.K., Rodriguez de la Fuente, L., Grundy, T.J., Fang, G., Valdes-Mora, F., and Gallego-Ortega, D. (2021). Advancements in 3D Cell Culture Systems for Personalizing AntiCancer Therapies. Front. Oncol., 11.","DOI":"10.3389\/fonc.2021.782766"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"1855","DOI":"10.1039\/C9BM02075F","article-title":"Hydrogel 3D in vitro tumor models for screening cell aggregation mediated drug response","volume":"8","author":"Monteiro","year":"2020","journal-title":"Biomater. Sci."},{"key":"ref_48","doi-asserted-by":"crossref","unstructured":"Abasalizadeh, F., Moghaddam, S.V., Alizadeh, E., Akbari, E., Kashani, E., Fazljou, S.M.B., Torbati, M., and Akbarzadeh, A. (2020). Alginate-based hydrogels as drug delivery vehicles in cancer treatment and their applications in wound dressing and 3D bioprinting. J. Biol. Eng., 14, Erratum in J. Biol. Eng. 2020, 14, 17. https:\/\/doi.org\/10.1186\/s13036-020-00239-0.","DOI":"10.1186\/s13036-020-0227-7"},{"key":"ref_49","doi-asserted-by":"crossref","unstructured":"Dai, X., Ma, C., Lan, Q., and Xu, T. (2016). 3D bioprinted glioma stem cells for brain tumor model and applications of drug susceptibility. Biofabrication, 8.","DOI":"10.1088\/1758-5090\/8\/4\/045005"},{"key":"ref_50","doi-asserted-by":"crossref","unstructured":"Jiang, T., Munguia-Lopez, J.G., Flores-Torres, S., Grant, J., Vijayakumar, S., Leon-Rodriguez, A., and Kinsella, J.M. (2017). Directing the self-assembly of tumour spheroids by bioprinting cellular heterogeneous models within alginate\/gelatin hydrogels. Sci. Rep., 7.","DOI":"10.1038\/s41598-017-04691-9"},{"key":"ref_51","first-page":"57826","article-title":"Bioprintable Alginate\/Gelatin Hydrogel 3D In Vitro Model Systems Induce Cell Spheroid Formation","volume":"137","author":"Jiang","year":"2018","journal-title":"J. Vis. Exp."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"5397","DOI":"10.1021\/acsbiomaterials.1c00920","article-title":"Bone Scaffolds: An Incorporation of Biomaterials, Cells, and Biofactors","volume":"7","author":"Bahraminasab","year":"2021","journal-title":"ACS Biomater. Sci. Eng."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"3567","DOI":"10.2174\/0929867324666170511123101","article-title":"Gelatin as Biomaterial for Tissue Engineering","volume":"23","author":"Echave","year":"2017","journal-title":"Curr. Pharm. Des."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"1063","DOI":"10.1039\/C4BM00070F","article-title":"Gelatin hydrogels formed by orthogonal thiol-norbornene photochemistry for cell encapsulation","volume":"2","author":"Shih","year":"2014","journal-title":"Biomater. Sci."},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"6977","DOI":"10.1039\/D3NJ04951E","article-title":"Design and synthesis of berberine loaded nano-hydroxyapatite\/gelatin scaffold for bone cancer treatment","volume":"48","author":"Khajavi","year":"2024","journal-title":"New J. Chem."},{"key":"ref_56","doi-asserted-by":"crossref","unstructured":"Cadamuro, F., Ardenti, V., Nicotra, F., and Russo, L. (2023). Alginate-Gelatin Self-Healing Hydrogel Produced via Static-Dynamic Crosslinking. Molecules, 28.","DOI":"10.3390\/molecules28062851"},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"4507","DOI":"10.1021\/cr3000169","article-title":"Biomimetic cell culture proteins as extracellular matrices for stem cell differentiation","volume":"112","author":"Higuchi","year":"2012","journal-title":"Chem. Rev."},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"47","DOI":"10.1016\/S0092-8674(00)80622-4","article-title":"Structural basis of collagen recognition by integrin alpha2beta1","volume":"101","author":"Emsley","year":"2000","journal-title":"Cell"},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"4590","DOI":"10.1021\/bm501386n","article-title":"Influence of amino acids, buffers, and ph on the \u03b3-irradiation-induced degradation of alginates","volume":"15","author":"Ulset","year":"2014","journal-title":"Biomacromolecules"},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"1302","DOI":"10.1021\/acsbiomaterials.1c00116","article-title":"Recent Development of Alginate-Based Materials and Their Versatile Functions in Biomedicine, Flexible Electronics, and Environmental Uses","volume":"7","author":"Teng","year":"2021","journal-title":"ACS Biomater. Sci. Eng."},{"key":"ref_61","doi-asserted-by":"crossref","first-page":"463","DOI":"10.1016\/j.jbiosc.2024.01.015","article-title":"Interpenetrating gelatin\/alginate mixed hydrogel: The simplest method to prepare an autoclavable scaffold","volume":"137","author":"Mori","year":"2024","journal-title":"J. Biosci. Bioeng."},{"key":"ref_62","first-page":"1377","article-title":"Effect of sunitinib derivatives on glioblastoma single-cell migration and 3D cell cultures","volume":"13","author":"Dabkeviciute","year":"2023","journal-title":"Am. J. Cancer Res."},{"key":"ref_63","doi-asserted-by":"crossref","unstructured":"Jo, H., Lee, S., Kim, M.H., Park, S., and Lee, S.Y. (2024). Recapitulating Glioma Stem Cell Niches Using 3D Spheroid Models for Glioblastoma Research. Biosensors, 14.","DOI":"10.3390\/bios14110539"},{"key":"ref_64","doi-asserted-by":"crossref","first-page":"10143","DOI":"10.1016\/j.biomaterials.2013.09.034","article-title":"Porous chitosan-hyaluronic acid scaffolds as a mimic of glioblastoma microenvironment ECM","volume":"34","author":"Florczyk","year":"2013","journal-title":"Biomaterials"},{"key":"ref_65","doi-asserted-by":"crossref","first-page":"833","DOI":"10.1038\/s41422-020-0338-1","article-title":"Three-dimensional bioprinted glioblastoma microenvironments model cellular dependencies and immune interactions","volume":"30","author":"Tang","year":"2020","journal-title":"Cell Res."},{"key":"ref_66","doi-asserted-by":"crossref","first-page":"132","DOI":"10.1016\/j.msec.2009.09.009","article-title":"Fabrication and characterization of bioactive silk fibroin\/wollastonite composite scaffolds","volume":"30","author":"Zhu","year":"2010","journal-title":"Mater. Sci. Eng."},{"key":"ref_67","doi-asserted-by":"crossref","unstructured":"Kapp, T.G., Rechenmacher, F., Neubauer, S., Maltsev, O.V., Cavalcanti-Adam, E.A., Zarka, R., Reuning, U., Notni, J., Wester, H.J., and Mas-Moruno, C. (2017). A Comprehensive Evaluation of the Activity and Selectivity Profile of Ligands for RGD-binding Integrins. Sci. Rep., 7.","DOI":"10.1038\/srep39805"},{"key":"ref_68","doi-asserted-by":"crossref","first-page":"148","DOI":"10.1007\/s10856-016-5763-9","article-title":"Evaluation of cell binding to collagen and gelatin: A study of the effect of 2D and 3D architecture and surface chemistry","volume":"27","author":"Davidenko","year":"2016","journal-title":"J. Mater. Sci. Mater. Med."},{"key":"ref_69","doi-asserted-by":"crossref","unstructured":"Chang, M.W., Lo, J.M., Juan, H.F., Chang, H.Y., and Chuang, C.Y. (2012). Combination of RGD compound and low-dose paclitaxel induces apoptosis in human glioblastoma cells. PLoS ONE, 7.","DOI":"10.1371\/journal.pone.0037935"},{"key":"ref_70","doi-asserted-by":"crossref","unstructured":"Isogai, R., Morio, H., Okamoto, A., Kitamura, K., and Furihata, T. (2022). Generation of a Human Conditionally Immortalized Cell-based Multicellular Spheroidal Blood-Brain Barrier Model for Permeability Evaluation of Macromolecules. Bio-Protocol, 12.","DOI":"10.21769\/BioProtoc.4465"},{"key":"ref_71","doi-asserted-by":"crossref","first-page":"120795","DOI":"10.1016\/j.ijpharm.2021.120795","article-title":"Nanomedicines functionalized with anti-EGFR ligands for active targeting in cancer therapy: Biological strategy, design and quality control","volume":"605","author":"Nguyen","year":"2021","journal-title":"Int. J. Pharm."},{"key":"ref_72","doi-asserted-by":"crossref","first-page":"100","DOI":"10.3389\/fonc.2017.00100","article-title":"Highly Specific and Effective Targeting of EGFRvIII-Positive Tumors with TandAb Antibodies","volume":"7","author":"Ellwanger","year":"2017","journal-title":"Front. Oncol."},{"key":"ref_73","doi-asserted-by":"crossref","unstructured":"Manzanares-Guzm\u00e1n, A., Alfonseca-Ladr\u00f3n de Guevara, A.C., Reza-Escobar, E., Burciaga-Flores, M., Canales-Aguirre, A., Esquivel-Sol\u00eds, H., Lugo-Fabres, P.H., and Camacho-Villegas, T.A. (2025). Isolation and Characterization of the First Antigen-Specific EGFRvIII vNAR from Freshwater Stingray (Potamotrygon spp.) as a Drug Carrier in Glioblastoma Cancer Cells. Int. J. Mol. Sci., 26.","DOI":"10.3390\/ijms26030876"},{"key":"ref_74","doi-asserted-by":"crossref","unstructured":"Malektaj, H., Drozdov, A.D., and deClaville Christiansen, J. (2023). Mechanical Properties of Alginate Hydrogels Cross-Linked with Multivalent Cations. Polymers, 15.","DOI":"10.3390\/polym15143012"},{"key":"ref_75","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"}],"container-title":["Gels"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2310-2861\/11\/4\/263\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T17:08:38Z","timestamp":1760029718000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2310-2861\/11\/4\/263"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,4,2]]},"references-count":75,"journal-issue":{"issue":"4","published-online":{"date-parts":[[2025,4]]}},"alternative-id":["gels11040263"],"URL":"https:\/\/doi.org\/10.3390\/gels11040263","relation":{},"ISSN":["2310-2861"],"issn-type":[{"value":"2310-2861","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,4,2]]}}}