{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,3]],"date-time":"2026-10-03T05:39:13Z","timestamp":1791005953789,"version":"4.1.0"},"reference-count":58,"publisher":"MDPI AG","issue":"19","license":[{"start":{"date-parts":[[2024,9,26]],"date-time":"2024-09-26T00:00:00Z","timestamp":1727308800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"JDZ\u2014now JBC","award":["EY005665"],"award-info":[{"award-number":["EY005665"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Communication between the different layers of the cornea (epithelium and stroma) is a complex, yet crucial element in the corneal healing process. Upon corneal injury, it has been reported that the bi-directional cross talk between the epithelium and stroma via the vesicular secretome, namely, extracellular vesicles (EVs), can lead to accelerated wound closure upon injury. However, the distinct protein markers of EVs derived from human corneal epithelial (HCE) cells, keratocytes (HCKs), fibroblasts (HCFs), and myofibroblasts (HCMs) remain poorly understood. All EVs were enriched for CD81 and showed increased expression levels of ITGAV and FN1 in HCM-EVs compared to HCE- and HCF-EVs. All EVs were negative for GM130 and showed minimal differences in biophysical properties (particle concentration, median particle size, and zeta potential). At the proteomic level, we show that HCM-EVs are enriched with proteins associated with fibrosis pathways, such as COL6A1, COL6A2, MMP1, MMP2, TIMP1, and TIMP2, compared to HCE-, HCK-, and HCF-EVs. Interestingly, HCE-EVs express proteins involved with the EIF-2 signaling pathway (stress-induced signals to regulate mRNA translation), such as RPS21, RALB, EIF3H, RALA, and others, compared to HCK-, HCF-, and HCM-EVs. In this study, we isolated EVs from cell-conditioned media from HCE, HCKs, HCFs, and HCMs and characterized their biophysical and protein composition by Western blot, nanoparticle tracking analysis, and proteomics. This study supports the view that EVs from the corneal epithelium and stroma have a distinct molecular composition and may provide novel protein markers to distinguish the difference between HCE-, HCK-, HCF-, and HCM-EVs.<\/jats:p>","DOI":"10.3390\/ijms251910338","type":"journal-article","created":{"date-parts":[[2024,9,26]],"date-time":"2024-09-26T04:05:46Z","timestamp":1727323546000},"page":"10338","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":13,"title":["Proteomic Characterization of Corneal Epithelial and Stromal Cell-Derived Extracellular Vesicles"],"prefix":"10.3390","volume":"25","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-5455-1049","authenticated-orcid":false,"given":"Vincent","family":"Yeung","sequence":"first","affiliation":[{"name":"Department of Ophthalmology, Schepens Eye Research Institute of Mass Eye and Ear, Harvard Medical School, Boston, MA 02114, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2674-0835","authenticated-orcid":false,"given":"Nikolay","family":"Boychev","sequence":"additional","affiliation":[{"name":"Department of Ophthalmology, Schepens Eye Research Institute of Mass Eye and Ear, Harvard Medical School, Boston, MA 02114, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6428-4696","authenticated-orcid":false,"given":"Levi N.","family":"Kanu","sequence":"additional","affiliation":[{"name":"Department of Ophthalmology, Schepens Eye Research Institute of Mass Eye and Ear, Harvard Medical School, Boston, MA 02114, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0003-9686-2608","authenticated-orcid":false,"given":"Veronica","family":"Ng","sequence":"additional","affiliation":[{"name":"Department of Ophthalmology, Schepens Eye Research Institute of Mass Eye and Ear, Harvard Medical School, Boston, MA 02114, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Amy E.","family":"Ross","sequence":"additional","affiliation":[{"name":"Department of Ophthalmology, Schepens Eye Research Institute of Mass Eye and Ear, Harvard Medical School, Boston, MA 02114, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Audrey E. 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Ophthalmol."},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Yeung, V., Zhang, T.C., Yuan, L., Parekh, M., Cortinas, J.A., Delavogia, E., Hutcheon, A.E.K., Guo, X., and Ciolino, J.B. (2022). Extracellular Vesicles Secreted by Corneal Myofibroblasts Promote Corneal Epithelial Cell Migration. Int. J. Mol. Sci., 23.","DOI":"10.3390\/ijms23063136"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"17","DOI":"10.1016\/j.preteyeres.2015.07.002","article-title":"Progress in corneal wound healing","volume":"49","author":"Ljubimov","year":"2015","journal-title":"Prog. Retin. Eye Res."},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Kowtharapu, B.S., Mur\u00edn, R., J\u00fcnemann, A.G.M., and Stachs, O. (2018). Role of Corneal Stromal Cells on Epithelial Cell Function during Wound Healing. Int. J. Mol. Sci., 19.","DOI":"10.3390\/ijms19020464"},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"McKay, T.B., Hutcheon, A.E.K., Zieske, J.D., and Ciolino, J.B. (2020). Extracellular Vesicles Secreted by Corneal Epithelial Cells Promote Myofibroblast Differentiation. Cells, 9.","DOI":"10.3390\/cells9051080"},{"key":"ref_6","first-page":"1465","article-title":"TGF-beta receptor types I and II are differentially expressed during corneal epithelial wound repair","volume":"42","author":"Zieske","year":"2001","journal-title":"Investig. Ophthalmol. Vis. Sci."},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Yeung, V., Sriram, S., Tran, J.A., Guo, X., Hutcheon, A.E.K., Zieske, J.D., Karamichos, D., and Ciolino, J.B. (2021). FAK Inhibition Attenuates Corneal Fibroblast Differentiation In Vitro. Biomolecules, 11.","DOI":"10.3390\/biom11111682"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"591","DOI":"10.1586\/14779072.2016.1147348","article-title":"Myofibroblast secretome and its auto-\/paracrine signaling","volume":"14","author":"Bomb","year":"2016","journal-title":"Expert. Rev. Cardiovasc. Ther."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"e007954","DOI":"10.1161\/JAHA.117.007954","article-title":"Examining the Paracrine Effects of Exosomes in Cardiovascular Disease and Repair","volume":"7","author":"Gartz","year":"2018","journal-title":"J. Am. Heart Assoc."},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Yeung, V., Willis, G.R., Taglauer, E., Mitsialis, S.A., and Kourembanas, S. (2019). Paving the Road for Mesenchymal Stem Cell-Derived Exosome Therapy in Bronchopulmonary Dysplasia and Pulmonary Hypertension. Stem Cell-Based Therapy for Lung Disease, Springer.","DOI":"10.1007\/978-3-030-29403-8_8"},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Habibi, A., Zarei-Behjani, Z., Falamarzi, K., Malekpour, M., Ebrahimi, F., Soleimani, M., Nejabat, M., Khosravi, A., Moayedfard, Z., and Pakbaz, S. (2023). Extracellular vesicles as a new horizon in the diagnosis and treatment of inflammatory eye diseases: A narrative review of the literature. Front. Immunol., 14.","DOI":"10.3389\/fimmu.2023.1097456"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"21","DOI":"10.1016\/j.jtos.2023.12.005","article-title":"Metabolic signatures of tear extracellular vesicles caused by herpes simplex keratitis","volume":"31","author":"Ma","year":"2024","journal-title":"Ocul. Surf."},{"key":"ref_13","first-page":"S73","article-title":"Fibroblast-myofibroblast crosstalk after exposure to mesenchymal stem cells secretome","volume":"11","author":"Angourani","year":"2018","journal-title":"Gastroenterol. Hepatol. Bed Bench"},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Han, K.-Y., Tran, J.A., Chang, J.-H., Azar, D.T., and Zieske, J.D. (2017). Potential role of corneal epithelial cell-derived exosomes in corneal wound healing and neovascularization. Sci. Rep., 7.","DOI":"10.1038\/srep40548"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"e12404","DOI":"10.1002\/jev2.12404","article-title":"Minimal information for studies of extracellular vesicles (MISEV2023): From basic to advanced approaches","volume":"13","author":"Welsh","year":"2024","journal-title":"J. Extracell. Vesicles"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1099","DOI":"10.1016\/S1097-2765(00)00108-8","article-title":"Regulated translation initiation controls stress-induced gene expression in mammalian cells","volume":"6","author":"Harding","year":"2000","journal-title":"Mol. Cell"},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Huang, Y.-H., I, C.-C., Kuo, C.-H., Hsu, Y.-Y., Lee, F.-T., Shi, G.-Y., Tseng, S.-H., and Wu, H.-L. (2015). Thrombomodulin promotes corneal epithelial wound healing. PLoS ONE, 10.","DOI":"10.1371\/journal.pone.0122491"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"108272","DOI":"10.1016\/j.exer.2020.108272","article-title":"Corneal myofibroblasts and fibrosis","volume":"201","author":"Wilson","year":"2020","journal-title":"Exp. Eye Res."},{"key":"ref_19","first-page":"614","article-title":"An SV40-immortalized human corneal epithelial cell line and its characterization","volume":"36","author":"Ohashi","year":"1995","journal-title":"Investig. Ophthalmol. Vis. Sci."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"565","DOI":"10.1007\/s00417-019-04563-0","article-title":"HCE-T cell line lacks cornea-specific differentiation markers compared to primary limbal epithelial cells and differentiated corneal epithelium","volume":"258","author":"Rubelowski","year":"2020","journal-title":"Graefe\u2019s Arch. Clin. Exp. Ophthalmol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"6390","DOI":"10.1167\/iovs.13-12547","article-title":"The corneal epithelial basement membrane: Structure, function, and disease","volume":"54","author":"Torricelli","year":"2013","journal-title":"Investig. Opthalmol. Vis. Sci."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"e9983900","DOI":"10.1155\/2021\/9983900","article-title":"Extracellular Vesicles in the Cornea: Insights from Other Tissues","volume":"2021","author":"McKay","year":"2021","journal-title":"Anal. Cell. Pathol."},{"key":"ref_23","first-page":"2102","article-title":"A comparison of stem cell-related gene expression in the progenitor-rich limbal epithelium and the differentiating central corneal epithelium","volume":"17","author":"Calonge","year":"2011","journal-title":"Mol. Vis."},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Albert, R., Ver\u00e9b, Z., Csom\u00f3s, K., Moe, M.C., Johnsen, E.O., Olstad, O.K., Nicolaissen, B., Rajnav\u00f6lgyi, E., F\u00e9s\u00fcs, L., and Berta, A. (2012). Cultivation and characterization of cornea limbal epithelial stem cells on lens capsule in animal material-free medium. PLoS ONE, 7.","DOI":"10.1371\/journal.pone.0047187"},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Lupasco, T., He, Z., Cassagne, M., Sagnial, T., Brion, L., Fourni\u00e9, P., Gain, P., Thuret, G., Allouche, M., and Malecaze, F. (2022). Corneal epithelium in keratoconus underexpresses active NRF2 and a subset of oxidative stress-related genes. PLoS ONE, 17.","DOI":"10.1371\/journal.pone.0273807"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"108206","DOI":"10.1016\/j.exer.2020.108206","article-title":"Keratin expression by corneal and limbal stem cells during development","volume":"200","author":"Kao","year":"2020","journal-title":"Exp. Eye Res."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"279","DOI":"10.1016\/j.jtos.2021.03.010","article-title":"A single cell atlas of human cornea that defines its development, limbal progenitor cells and their interactions with the immune cells","volume":"21","author":"Collin","year":"2021","journal-title":"Ocul. Surf."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"e2204134120","DOI":"10.1073\/pnas.2204134120","article-title":"Dry eye disease in mice activates adaptive corneal epithelial regeneration distinct from constitutive renewal in homeostasis","volume":"120","author":"Lin","year":"2023","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"434","DOI":"10.1080\/19336918.2016.1245264","article-title":"Orphan G protein-coupled receptor GPRC5A modulates integrin \u03b21-mediated epithelial cell adhesion","volume":"11","author":"Bulanova","year":"2017","journal-title":"Cell Adhes. Migr."},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"Liu, B., Yang, H., Pilarsky, C., and Weber, G.F. (2018). The Effect of GPRC5a on the Proliferation, Migration Ability, Chemotherapy Resistance, and Phosphorylation of GSK-3\u03b2 in Pancreatic Cancer. Int. J. Mol. Sci., 19.","DOI":"10.3390\/ijms19071870"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"1369","DOI":"10.1002\/ijc.32554","article-title":"GPRC5A facilitates cell proliferation through cell cycle regulation and correlates with bone metastasis in prostate cancer","volume":"146","author":"Sawada","year":"2020","journal-title":"Int. J. Cancer"},{"key":"ref_32","first-page":"63","article-title":"Recessively Inherited Deficiency of Secreted WFDC2 (HE4) Causes Nasal Polyposis and Bronchiectasis","volume":"210","author":"Dougherty","year":"2024","journal-title":"Am. J. Respir. Crit. Care Med."},{"key":"ref_33","first-page":"348","article-title":"Collagen VI disorders: Insights on form and function in the extracellular matrix and beyond","volume":"71","author":"Bateman","year":"2018","journal-title":"Matrix Biol."},{"key":"ref_34","doi-asserted-by":"crossref","unstructured":"Zanotti, S., Magri, F., Salani, S., Napoli, L., Ripolone, M., Ronchi, D., Fortunato, F., Ciscato, P., Velardo, D., and D\u2019aNgelo, M.G. (2023). Extracellular Matrix Disorganization and Sarcolemmal Alterations in COL6-Related Myopathy Patients with New Variants of COL6 Genes. Int. J. Mol. Sci., 24.","DOI":"10.3390\/ijms24065551"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"315","DOI":"10.1007\/s004020050417","article-title":"Immunohistochemical localization of collagen VI in arthrofibrosis","volume":"119","author":"Zeichen","year":"1999","journal-title":"Arch. Orthop. Trauma Surg."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"789","DOI":"10.1002\/hep.22099","article-title":"Relationship of serum fibrosis markers with liver fibrosis stage and collagen content in patients with advanced chronic hepatitis C","volume":"47","author":"Fontana","year":"2008","journal-title":"Hepatology"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"20753","DOI":"10.1073\/pnas.2004281117","article-title":"Identifying collagen VI as a target of fibrotic diseases regulated by CREBBP\/EP300","volume":"117","author":"Williams","year":"2020","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_38","doi-asserted-by":"crossref","unstructured":"Jessen, H., Hoyer, N., Prior, T.S., Frederiksen, P., R\u00f8nnow, S.R., Karsdal, M.A., Leeming, D.J., Bendstrup, E., Sand, J.M.B., and Shaker, S.B. (2021). Longitudinal serological assessment of type VI collagen turnover is related to progression in a real-world cohort of idiopathic pulmonary fibrosis. BMC Pulm. Med., 21.","DOI":"10.1186\/s12890-021-01684-3"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"5050","DOI":"10.1111\/jcmm.13787","article-title":"CXCL6-EGFR-induced Kupffer cells secrete TGF-\u03b21 promoting hepatic stellate cell activation via the SMAD2\/BRD4\/C-MYC\/EZH2 pathway in liver fibrosis","volume":"22","author":"Cai","year":"2018","journal-title":"J. Cell. Mol. Med."},{"key":"ref_40","doi-asserted-by":"crossref","unstructured":"Kersey, A.L., Cheng, D.Y., Deo, K.A., Dubell, C.R., Wang, T.-C., Jaiswal, M.K., Kim, M.H., Murali, A., Hargett, S.E., and Mallick, S. (2024). Stiffness assisted cell-matrix remodeling trigger 3D mechanotransduction regulatory programs. Biomaterials, 306.","DOI":"10.1016\/j.biomaterials.2024.122473"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"1375","DOI":"10.1158\/2159-8290.CD-24-0002","article-title":"The Interplay between Extracellular Matrix Remodeling and Cancer Therapeutics","volume":"14","author":"Prakash","year":"2024","journal-title":"Cancer Discov."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"3413","DOI":"10.1021\/pr400270q","article-title":"Mapping proteolytic processing in the secretome of gastric cancer-associated myofibroblasts reveals activation of MMP-1, MMP-2, and MMP-3","volume":"12","author":"Holmberg","year":"2013","journal-title":"J. Proteome Res."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"463","DOI":"10.1016\/j.matbio.2017.12.012","article-title":"Matrix metalloproteinases and liver fibrosis (translational aspects)","volume":"68","author":"Roeb","year":"2018","journal-title":"Matrix Biol."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"308","DOI":"10.1016\/j.addr.2020.03.002","article-title":"An emerging focus on lipids in extracellular vesicles","volume":"159","author":"Skotland","year":"2020","journal-title":"Adv. Drug Deliv. Rev."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"e12260","DOI":"10.1002\/jev2.12260","article-title":"Understanding the extracellular vesicle surface for clinical molecular biology","volume":"11","author":"Hallal","year":"2022","journal-title":"J. Extracell. Vesicles"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"D1694","DOI":"10.1093\/nar\/gkad1007","article-title":"Vesiclepedia 2024: An extracellular vesicles and extracellular particles repository","volume":"52","author":"Chitti","year":"2024","journal-title":"Nucleic Acids Res."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"e12481","DOI":"10.1002\/jev2.12481","article-title":"Extracellular vesicles carry transcriptional \u2018dark matter\u2019 revealing tissue-specific information","volume":"13","author":"Dogra","year":"2024","journal-title":"J. Extracell. Vesicles"},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"1908","DOI":"10.1083\/jcb.201811173","article-title":"Rab5-dependent autophagosome closure by ESCRT","volume":"218","author":"Zhou","year":"2019","journal-title":"J. Cell Biol."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"657","DOI":"10.1038\/s41418-020-00610-0","article-title":"Targeting the ESCRT-III component CHMP2A for noncanonical Caspase-8 activation on autophagosomal membranes","volume":"28","author":"Hattori","year":"2021","journal-title":"Cell Death Differ."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"13918","DOI":"10.1074\/jbc.275.18.13918","article-title":"Fibroblast growth factor-2 promotes keratan sulfate proteoglycan expression by keratocytes in vitro","volume":"275","author":"Long","year":"2000","journal-title":"J. Biol. Chem."},{"key":"ref_51","doi-asserted-by":"crossref","unstructured":"Foster, J.W., Gouveia, R.M., and Connon, C.J. (2015). Low-glucose enhances keratocyte-characteristic phenotype from corneal stromal cells in serum-free conditions. Sci. Rep., 5.","DOI":"10.1038\/srep10839"},{"key":"ref_52","doi-asserted-by":"crossref","unstructured":"Yusoff, N.Z.B.M., Riau, A.K., Yam, G.H.F., Halim, N.S.H.B., and Mehta, J.S. (2022). Isolation and Propagation of Human Corneal Stromal Keratocytes for Tissue Engineering and Cell Therapy. Cells, 11.","DOI":"10.3390\/cells11010178"},{"key":"ref_53","first-page":"1658","article-title":"Proteoglycan synthesis by bovine keratocytes and corneal fibroblasts: Maintenance of the keratocyte phenotype in culture","volume":"40","author":"Beales","year":"1999","journal-title":"Investig. Ophthalmol. Vis. Sci."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"6612","DOI":"10.1167\/iovs.13-12861","article-title":"TGF-\u03b23 Stimulates Stromal Matrix Assembly by Human Corneal Keratocyte-Like Cells","volume":"54","author":"Karamichos","year":"2013","journal-title":"Investig. Opthalmol. Vis. Sci."},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"2705","DOI":"10.1002\/dvdy.21606","article-title":"Human Primary Corneal Fibroblasts Synthesize and Deposit Proteoglycans in Long-Term 3-D Cultures","volume":"237","author":"Ren","year":"2008","journal-title":"Dev. Dyn."},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"8547","DOI":"10.1039\/C8NR02417K","article-title":"Rab35-dependent extracellular nanovesicles are required for induction of tumour supporting stroma","volume":"10","author":"Yeung","year":"2018","journal-title":"Nanoscale"},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"19671","DOI":"10.3402\/jev.v2i0.19671","article-title":"Extracellular vesicle sizing and enumeration by nanoparticle tracking analysis","volume":"2","author":"Gardiner","year":"2013","journal-title":"J. Extracell. Vesicles"},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"3","DOI":"10.1002\/0471143030.cb0322s30","article-title":"Isolation and characterization of exosomes from cell culture supernatants and biological fluids","volume":"30","author":"Thery","year":"2006","journal-title":"Curr. Protoc. Cell Biol."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/25\/19\/10338\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T16:03:25Z","timestamp":1760112205000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/25\/19\/10338"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,9,26]]},"references-count":58,"journal-issue":{"issue":"19","published-online":{"date-parts":[[2024,10]]}},"alternative-id":["ijms251910338"],"URL":"https:\/\/doi.org\/10.3390\/ijms251910338","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,9,26]]}}}