{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T13:19:10Z","timestamp":1785417550447,"version":"3.56.0"},"reference-count":56,"publisher":"MDPI AG","issue":"8","license":[{"start":{"date-parts":[[2026,7,23]],"date-time":"2026-07-23T00:00:00Z","timestamp":1784764800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100004410","name":"Scientific and Technological Research Council of Turkey","doi-asserted-by":"publisher","award":["122Z739"],"award-info":[{"award-number":["122Z739"]}],"id":[{"id":"10.13039\/501100004410","id-type":"DOI","asserted-by":"publisher"}]},{"award":["122Z739"],"award-info":[{"award-number":["122Z739"]}],"id":[{"id":"https:\/\/ror.org\/04w9kkr77","id-type":"ROR","asserted-by":"publisher"}]},{"name":"Recep Tayyip Erdo\u011fan University Development Foundation","award":["020260070080462"],"award-info":[{"award-number":["020260070080462"]}]},{"award":["020260070080462"],"award-info":[{"award-number":["020260070080462"]}],"id":[{"id":"https:\/\/ror.org\/0468j1635","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biomolecules"],"abstract":"<jats:p>Glioblastoma (GBM) is the most aggressive primary brain tumor and remains associated with poor clinical outcomes despite advances in surgical and adjuvant therapies. The tumor microenvironment, particularly extracellular matrix (ECM) interactions, plays a crucial role in regulating glioblastoma progression, cellular plasticity, and therapeutic resistance. Therefore, physiologically relevant three-dimensional (3D) models are needed to better recapitulate GBM biology. In this study, we investigated the effects of ECM-functionalized polyacrylonitrile\/coumarin-500 (PAN\/C500) nanofiber scaffolds on the phenotype of LN-18 and U-87 MG glioblastoma cells cultured under 3D conditions. Cytoskeletal organization was assessed by phalloidin staining and live-cell vimentin imaging, while epithelial\u2013mesenchymal transition (EMT)-associated proteins and stemness-related markers were analyzed by Western blotting. ECM-functionalized 3D PAN\/C500 scaffolds promoted significant cytoskeletal remodeling, altered EMT-associated protein expression, and increased the expression of stemness-associated proteins, particularly SOX2, NANOG, and Nestin, compared with conventional 2D cultures. These responses were accompanied by cell line-dependent phenotypic adaptations, indicating that the engineered microenvironment influences glioblastoma cell behavior. This platform may serve as a valuable model for investigating glioblastoma biology and microenvironment-associated molecular adaptations in vitro.<\/jats:p>","DOI":"10.3390\/biom16081077","type":"journal-article","created":{"date-parts":[[2026,7,23]],"date-time":"2026-07-23T09:11:26Z","timestamp":1784797886000},"page":"1077","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Three-Dimensional ECM-Functionalized PAN\/C500 Nanofiber Scaffolds Induce Cytoskeletal Remodeling and Stemness-Associated Molecular Changes in Glioblastoma Cells"],"prefix":"10.3390","volume":"16","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-7246-2592","authenticated-orcid":false,"given":"Ihsan","family":"Nalkiran","sequence":"first","affiliation":[{"name":"Department of Medical Biology, Faculty of Medicine, Recep Tayyip Erdogan University, Rize 53100, T\u00fcrkiye"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1115-2005","authenticated-orcid":false,"given":"Hatice","family":"Sevim Nalkiran","sequence":"additional","affiliation":[{"name":"Department of Medical Biology, Faculty of Medicine, Recep Tayyip Erdogan University, Rize 53100, T\u00fcrkiye"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6544-6271","authenticated-orcid":false,"given":"Derya Bal","family":"Altuntas","sequence":"additional","affiliation":[{"name":"Department of Bioengineering, Faculty of Engineering and Architecture, Recep Tayyip Erdogan University, Rize 53100, T\u00fcrkiye"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1524-3342","authenticated-orcid":false,"given":"Atilla Eren","family":"Mamuk","sequence":"additional","affiliation":[{"name":"Department of Physics, Faculty of Science, Mugla Sitki Kocman University, Mu\u011fla 48100, T\u00fcrkiye"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Cagdas","family":"Kocak","sequence":"additional","affiliation":[{"name":"Department of Physics, Faculty of Science, Mugla Sitki Kocman University, Mu\u011fla 48100, T\u00fcrkiye"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ebiha","family":"Can","sequence":"additional","affiliation":[{"name":"Department of Medical Biology, Faculty of Medicine, Recep Tayyip Erdogan University, Rize 53100, T\u00fcrkiye"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sema","family":"Aslan","sequence":"additional","affiliation":[{"name":"Department of Chemistry, Faculty of Science, Mugla Sitki Kocman University, Mu\u011fla 48100, T\u00fcrkiye"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,7,23]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"iv1","DOI":"10.1093\/neuonc\/noaa200","article-title":"CBTRUS Statistical Report: Primary Brain and Other Central Nervous System Tumors Diagnosed in the United States in 2013\u20132017","volume":"22","author":"Ostrom","year":"2020","journal-title":"Neuro-Oncology"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"987","DOI":"10.1056\/NEJMoa043330","article-title":"Radiotherapy plus concomitant and adjuvant temozolomide for glioblastoma","volume":"352","author":"Stupp","year":"2005","journal-title":"N. Engl. J. Med."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"3029","DOI":"10.1002\/cncr.33587","article-title":"The ever-increasing importance of cancer as a leading cause of premature death worldwide","volume":"127","author":"Bray","year":"2021","journal-title":"Cancer"},{"key":"ref_4","first-page":"299","article-title":"Management of glioblastoma: State of the art and future directions","volume":"70","author":"Tan","year":"2020","journal-title":"CA Cancer J. Clin."},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Manini, I., Caponnetto, F., Bartolini, A., Ius, T., Mariuzzi, L., Di Loreto, C., Beltrami, A.P., and Cesselli, D. (2018). Role of Microenvironment in Glioma Invasion: What We Learned from In Vitro Models. Int. J. Mol. Sci., 19.","DOI":"10.3390\/ijms19010147"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"761","DOI":"10.7150\/jca.17648","article-title":"Role of tumor microenvironment in tumorigenesis","volume":"8","author":"Wang","year":"2017","journal-title":"J. Cancer"},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Huijbers, I.J., Iravani, M., Popov, S., Robertson, D., Al-Sarraj, S., Jones, C., and Isacke, C.M. (2010). A role for fibrillar collagen deposition and the collagen internalization receptor endo180 in glioma invasion. PLoS ONE, 5.","DOI":"10.1371\/journal.pone.0009808"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"265","DOI":"10.1146\/annurev-cellbio-092910-154013","article-title":"Transmembrane collagen receptors","volume":"27","author":"Leitinger","year":"2011","journal-title":"Annu. Rev. Cell Dev. Biol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"687","DOI":"10.1038\/444687a","article-title":"Cancer: Stem cells and brain tumours","volume":"444","author":"Dirks","year":"2006","journal-title":"Nature"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"40","DOI":"10.1634\/stemcells.2008-0493","article-title":"SOX2 silencing in glioblastoma tumor-initiating cells causes stop of proliferation and loss of tumorigenicity","volume":"27","author":"Gangemi","year":"2009","journal-title":"Stem Cells"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"37","DOI":"10.1038\/s41392-021-00857-0","article-title":"Sox2 induces glioblastoma cell stemness and tumor propagation by repressing TET2 and deregulating 5hmC and 5mC DNA modifications","volume":"7","author":"Johnson","year":"2022","journal-title":"Signal Transduct. Target. Ther."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"157","DOI":"10.1016\/j.bbrc.2009.02.156","article-title":"Embryonic stem cell markers expression in cancers","volume":"383","author":"Schoenhals","year":"2009","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"643","DOI":"10.1016\/S0092-8674(03)00392-1","article-title":"Functional expression cloning of Nanog, a pluripotency sustaining factor in embryonic stem cells","volume":"113","author":"Chambers","year":"2003","journal-title":"Cell"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"663","DOI":"10.1016\/j.cell.2006.07.024","article-title":"Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors","volume":"126","author":"Takahashi","year":"2006","journal-title":"Cell"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"1917","DOI":"10.1126\/science.1151526","article-title":"Induced pluripotent stem cell lines derived from human somatic cells","volume":"318","author":"Yu","year":"2007","journal-title":"Science"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"2659","DOI":"10.1038\/emboj.2010.137","article-title":"NANOG regulates glioma stem cells and is essential in vivo acting in a cross-functional network with GLI1 and p53","volume":"29","author":"Zbinden","year":"2010","journal-title":"EMBO J."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"2281","DOI":"10.1016\/j.celrep.2016.02.034","article-title":"Plasticity between Epithelial and Mesenchymal States Unlinks EMT from Metastasis-Enhancing Stem Cell Capacity","volume":"14","author":"Beerling","year":"2016","journal-title":"Cell Rep."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"151523","DOI":"10.1016\/j.ejcb.2025.151523","article-title":"Vimentin in the tumor microenvironment: Orchestrating invasion, immunity, and metabolism","volume":"104","author":"Parvanian","year":"2025","journal-title":"Eur. J. Cell Biol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1038\/s12276-025-01608-9","article-title":"Roles of cytoskeleton in metastasis: From its mechanism to therapeutic strategies","volume":"58","author":"Lim","year":"2026","journal-title":"Exp. Mol. Med."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"32","DOI":"10.1186\/s12964-021-00713-2","article-title":"The epithelial-mesenchymal transition and the cytoskeleton in bioengineered systems","volume":"19","author":"Leggett","year":"2021","journal-title":"Cell Commun. Signal."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"3","DOI":"10.1016\/j.jbiotec.2010.01.012","article-title":"Multicellular tumor spheroids: An underestimated tool is catching up again","volume":"148","author":"Hirschhaeuser","year":"2010","journal-title":"J. Biotechnol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"4947","DOI":"10.1039\/C7PY00559H","article-title":"Multicellular tumor spheroids: A relevant 3D model for the in vitro preclinical investigation of polymer nanomedicines","volume":"8","author":"Lazzari","year":"2017","journal-title":"Polym. Chem."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"391","DOI":"10.1016\/j.ccr.2006.03.030","article-title":"Tumor stem cells derived from glioblastomas cultured in bFGF and EGF more closely mirror the phenotype and genotype of primary tumors than do serum-cultured cell lines","volume":"9","author":"Lee","year":"2006","journal-title":"Cancer Cell"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"7408","DOI":"10.1016\/j.biomaterials.2013.06.024","article-title":"Regulation of glioma cell phenotype in 3D matrices by hyaluronic acid","volume":"34","author":"Pedron","year":"2013","journal-title":"Biomaterials"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"56904","DOI":"10.18632\/oncotarget.10885","article-title":"A three-dimensional collagen scaffold cell culture system for screening anti-glioma therapeutics","volume":"7","author":"Lv","year":"2016","journal-title":"Oncotarget"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"62","DOI":"10.1016\/j.biomaterials.2015.11.031","article-title":"Collaboration of 3D context and extracellular matrix in the development of glioma stemness in a 3D model","volume":"78","author":"Ma","year":"2016","journal-title":"Biomaterials"},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"Rosellini, E., and Cascone, M.G. (2025). Scaffolds Mimicking the Tumor Microenvironment for In Vitro Malignancy Models. Biomimetics, 10.","DOI":"10.3390\/biomimetics10100695"},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Anjum, S., Rahman, F., Pandey, P., Arya, D.K., Alam, M., Rajinikanth, P.S., and Ao, Q. (2022). Electrospun Biomimetic Nanofibrous Scaffolds: A Promising Prospect for Bone Tissue Engineering and Regenerative Medicine. Int. J. Mol. Sci., 23.","DOI":"10.3390\/ijms23169206"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"102385","DOI":"10.1016\/j.mtbio.2025.102385","article-title":"Multifunctional nanoplatforms for tumor microenvironment remodeling: Toward precision and intelligent cancer therapy","volume":"35","author":"Deng","year":"2025","journal-title":"Mater. Today Bio"},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"Santillan, V.E., Nagam Hanumantharao, S., Bule, S., Shrestha, R.M., Rodzik, C., Mendoza Estrada, A., Farias, S.L., Tanasova, M., and Rao, S. (2026). Electrospun Nanofiber Scaffolds for In Vitro 3D Tissue Engineering. Fibers, 14.","DOI":"10.3390\/fib14060065"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"12644","DOI":"10.1038\/s41598-026-39640-y","article-title":"Electrochemically optimized multi-component polyacrylonitrile nanofiber scaffolds as a platform for three-dimensional glioblastoma cell culture","volume":"16","author":"Kurt","year":"2026","journal-title":"Sci. Rep."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"139006","DOI":"10.1016\/j.snb.2025.139006","article-title":"Three-dimensional cytosensor for glioblastoma cell analysis using coumarin-integrated nanofiber scaffolds on ITO electrodes","volume":"449","author":"Kurt","year":"2026","journal-title":"Sens. Actuators B Chem."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"571","DOI":"10.1007\/s00280-008-0881-9","article-title":"Cancer and the tumor microenvironment: A review of an essential relationship","volume":"63","author":"Mbeunkui","year":"2009","journal-title":"Cancer Chemother. Pharmacol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"2115","DOI":"10.1021\/mp5000828","article-title":"Bioengineered 3D brain tumor model to elucidate the effects of matrix stiffness on glioblastoma cell behavior using PEG-based hydrogels","volume":"11","author":"Wang","year":"2014","journal-title":"Mol. Pharm."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"471","DOI":"10.1016\/j.ccell.2020.03.007","article-title":"Intratumor Heterogeneity: The Rosetta Stone of Therapy Resistance","volume":"37","author":"Marusyk","year":"2020","journal-title":"Cancer Cell"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"575","DOI":"10.1158\/2159-8290.CD-20-1474","article-title":"Diffuse Glioma Heterogeneity and Its Therapeutic Implications","volume":"11","author":"Nicholson","year":"2021","journal-title":"Cancer Discov."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"2020","DOI":"10.3389\/fmolb.2020.00033","article-title":"Is It Time to Start Transitioning From 2D to 3D Cell Culture?","volume":"7","author":"Jensen","year":"2020","journal-title":"Front. Mol. Biosci."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"524","DOI":"10.3390\/macromol3030031","article-title":"Electrospun Scaffolds for Tissue Engineering: A Review","volume":"3","author":"Bucio","year":"2023","journal-title":"Macromol"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"1953861","DOI":"10.1155\/2022\/1953861","article-title":"Recent Applications of Electrospun Nanofibrous Scaffold in Tissue Engineering","volume":"2022","author":"Owida","year":"2022","journal-title":"Appl. Bionics Biomech."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"463","DOI":"10.4103\/jomfp.JOMFP_132_16","article-title":"Cancer stem cells: An insight","volume":"21","author":"Moharil","year":"2017","journal-title":"J. Oral Maxillofac. Pathol."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"25","DOI":"10.1095\/biolreprod.114.121368","article-title":"Dynamic reciprocity between cells and their microenvironment in reproduction","volume":"92","author":"Thorne","year":"2015","journal-title":"Biol. Reprod."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"4529","DOI":"10.1038\/s41467-019-12420-1","article-title":"3D extracellular matrix microenvironment in bioengineered tissue models of primary pediatric and adult brain tumors","volume":"10","author":"Sood","year":"2019","journal-title":"Nat. Commun."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"7011","DOI":"10.1158\/0008-5472.CAN-04-1364","article-title":"Isolation and characterization of tumorigenic, stem-like neural precursors from human glioblastoma","volume":"64","author":"Galli","year":"2004","journal-title":"Cancer Res."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"947","DOI":"10.1016\/j.cell.2005.08.020","article-title":"Core transcriptional regulatory circuitry in human embryonic stem cells","volume":"122","author":"Boyer","year":"2005","journal-title":"Cell"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"631","DOI":"10.1016\/S0092-8674(03)00393-3","article-title":"The homeoprotein Nanog is required for maintenance of pluripotency in mouse epiblast and ES cells","volume":"113","author":"Mitsui","year":"2003","journal-title":"Cell"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"722","DOI":"10.1016\/j.cell.2009.07.039","article-title":"Nanog is the gateway to the pluripotent ground state","volume":"138","author":"Silva","year":"2009","journal-title":"Cell"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"2177","DOI":"10.1007\/s00018-018-2794-z","article-title":"Nestin-expressing progenitor cells: Function, identity and therapeutic implications","volume":"75","author":"Bernal","year":"2018","journal-title":"Cell. Mol. Life Sci."},{"key":"ref_48","first-page":"91","article-title":"Neuroepithelial stem cell marker nestin regulates the migration, invasion and growth of human gliomas","volume":"26","author":"Ishiwata","year":"2011","journal-title":"Oncol. Rep."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"6281","DOI":"10.1158\/0008-5472.CAN-08-0094","article-title":"Oct-3\/4 expression reflects tumor progression and regulates motility of bladder cancer cells","volume":"68","author":"Chang","year":"2008","journal-title":"Cancer Res."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"165","DOI":"10.1016\/j.cub.2006.11.033","article-title":"HEDGEHOG-GLI1 signaling regulates human glioma growth, cancer stem cell self-renewal, and tumorigenicity","volume":"17","author":"Clement","year":"2007","journal-title":"Curr. Biol."},{"key":"ref_51","first-page":"2275","article-title":"PARP-1 inhibition sensitizes temozolomide-treated glioblastoma cell lines and decreases drug resistance independent of MGMT activity and PTEN proficiency","volume":"44","author":"Montaldi","year":"2020","journal-title":"Oncol. Rep."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"425","DOI":"10.1038\/s41392-021-00828-5","article-title":"Targeting PI3K\/Akt signal transduction for cancer therapy","volume":"6","author":"He","year":"2021","journal-title":"Signal Transduct. Target. Ther."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"666","DOI":"10.1038\/cdd.2010.139","article-title":"PTEN status switches cell fate between premature senescence and apoptosis in glioma exposed to ionizing radiation","volume":"18","author":"Lee","year":"2011","journal-title":"Cell Death Differ."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"43452","DOI":"10.18632\/oncotarget.6171","article-title":"Elucidating the cancer-specific genetic alteration spectrum of glioblastoma derived cell lines from whole exome and RNA sequencing","volume":"6","author":"Patil","year":"2015","journal-title":"Oncotarget"},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"3149","DOI":"10.1021\/pr200210w","article-title":"Secretome Signature of Invasive Glioblastoma Multiforme","volume":"10","author":"Formolo","year":"2011","journal-title":"J. Proteome Res."},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"14612","DOI":"10.1038\/s41598-019-51108-w","article-title":"A novel 3D nanofibre scaffold conserves the plasticity of glioblastoma stem cell invasion by regulating galectin-3 and integrin-\u03b21 expression","volume":"9","author":"Saleh","year":"2019","journal-title":"Sci. Rep."}],"container-title":["Biomolecules"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2218-273X\/16\/8\/1077\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,7,23]],"date-time":"2026-07-23T09:46:28Z","timestamp":1784799988000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2218-273X\/16\/8\/1077"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,7,23]]},"references-count":56,"journal-issue":{"issue":"8","published-online":{"date-parts":[[2026,8]]}},"alternative-id":["biom16081077"],"URL":"https:\/\/doi.org\/10.3390\/biom16081077","relation":{},"ISSN":["2218-273X"],"issn-type":[{"value":"2218-273X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,7,23]]}}}