{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,12]],"date-time":"2026-08-12T11:36:58Z","timestamp":1786534618628,"version":"build-2736575974"},"reference-count":34,"publisher":"MDPI AG","issue":"16","license":[{"start":{"date-parts":[[2026,8,12]],"date-time":"2026-08-12T00:00:00Z","timestamp":1786492800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["82272171"],"award-info":[{"award-number":["82272171"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["31971279"],"award-info":[{"award-number":["31971279"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"award":["82272171"],"award-info":[{"award-number":["82272171"]}],"id":[{"id":"https:\/\/ror.org\/01h0zpd94","id-type":"ROR","asserted-by":"publisher"}]},{"award":["31971279"],"award-info":[{"award-number":["31971279"]}],"id":[{"id":"https:\/\/ror.org\/01h0zpd94","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Int. J. Mol. Sci.","IJMS"],"abstract":"<jats:p>N-methyl-D-aspartate (NMDA) excitotoxicity drives mitochondrial dysfunction and retinal ganglion cell (RGC) loss in blinding retinal disorders. Ciliary neurotrophic factor (CNTF) is neuroprotective, but its short half-life limits long-term therapy. Here, we investigated whether mitochondrial homeostasis mediates the sustained protection of a CNTF-loaded chitosan hydrogel. In vitro, free CNTF and the hydrogel equally protected RGCs against NMDA, effects completely abolished by the mitochondrial uncoupler carbonyl cyanide m-chlorophenyl hydrazone (CCCP). In vivo, free CNTF provided only transient rescue, whereas the hydrogel sustained RGC survival, mitochondrial integrity, and visual function for 28 days, with persistent upregulation of mitochondrial biogenesis genes. Adeno-associated virus-mediated DRP1 overexpression-induced mitochondrial fission fully reversed the hydrogel\u2019s benefits, mirroring CCCP inhibition. Collectively, intact mitochondrial homeostasis is essential for the long-term neuroprotection of the CNTF-chitosan hydrogel, which extends CNTF retention without altering its mitochondrial-dependent mechanism. This hydrogel represents a promising long-acting treatment for retinal excitotoxicity.<\/jats:p>","DOI":"10.3390\/ijms27167200","type":"journal-article","created":{"date-parts":[[2026,8,12]],"date-time":"2026-08-12T11:02:11Z","timestamp":1786532531000},"page":"7200","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Mitochondrial Homeostasis Mediates the Sustained Neuroprotection of CNTF-Chitosan Hydrogel Against NMDA-Induced Retinal Ganglion Cell Excitotoxicity"],"prefix":"10.3390","volume":"27","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-0756-834X","authenticated-orcid":false,"given":"Huiting","family":"Jiang","sequence":"first","affiliation":[{"name":"Department of Neurobiology, School of Basic Medical Sciences, Capital Medical University, Beijing 100069, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xunhui","family":"Guo","sequence":"additional","affiliation":[{"name":"Department of Neurobiology, School of Basic Medical Sciences, Capital Medical University, Beijing 100069, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuanyuan","family":"Bai","sequence":"additional","affiliation":[{"name":"Department of Neurobiology, School of Basic Medical Sciences, Capital Medical University, Beijing 100069, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xinyue","family":"Ma","sequence":"additional","affiliation":[{"name":"Department of Neurobiology, School of Basic Medical Sciences, Capital Medical University, Beijing 100069, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hongmei","family":"Duan","sequence":"additional","affiliation":[{"name":"Department of Neurobiology, School of Basic Medical Sciences, Capital Medical University, Beijing 100069, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaoguang","family":"Li","sequence":"additional","affiliation":[{"name":"Department of Neurobiology, School of Basic Medical Sciences, Capital Medical University, Beijing 100069, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhaoyang","family":"Yang","sequence":"additional","affiliation":[{"name":"Department of Neurobiology, School of Basic Medical Sciences, Capital Medical University, Beijing 100069, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,8,12]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Kashihara, T., Morita, Y., Hatta, M., Inoue, S., Suzuki, Y., Morita, A., and Nakahara, T. (2024). YAP activation in M\u00fcller cells protects against NMDA-induced retinal ganglion cell injury by regulating Bcl-xL expression. Front. Pharmacol., 15.","DOI":"10.3389\/fphar.2024.1446521"},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Feng, K.M., Tsung, T.-H., Chen, Y.-H., and Lu, D.-W. (2023). The Role of Retinal Ganglion Cell Structure and Function in Glaucoma. Cells, 12.","DOI":"10.3390\/cells12242797"},{"key":"ref_3","doi-asserted-by":"crossref","unstructured":"Boccuni, I., and Fairless, R. (2022). Retinal Glutamate Neurotransmission: From Physiology to Pathophysiological Mechanisms of Retinal Ganglion Cell Degeneration. Life, 12.","DOI":"10.3390\/life12050638"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"28","DOI":"10.1186\/s40478-025-01942-z","article-title":"Disruption of mitochondrial homeostasis and permeability transition pore opening in OPA1 iPSC-derived retinal ganglion cells","volume":"13","author":"Whitehead","year":"2025","journal-title":"Acta Neuropathol. Commun."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"3834","DOI":"10.1038\/s41598-024-54075-z","article-title":"Ripa-56 protects retinal ganglion cells in glutamate-induced retinal excitotoxic model of glaucoma","volume":"14","author":"Feng","year":"2024","journal-title":"Sci. Rep."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"262","DOI":"10.1186\/s12974-022-02626-4","article-title":"GSK872 and necrostatin-1 protect retinal ganglion cells against necroptosis through inhibition of RIP1\/RIP3\/MLKL pathway in glutamate-induced retinal excitotoxic model of glaucoma","volume":"19","author":"Liu","year":"2022","journal-title":"J. Neuroinflamm."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"125","DOI":"10.3341\/kjo.2024.0148","article-title":"Neuroprotective Mechanisms of Ciliary Neurotrophic Factor in Retinal Ganglion Cells: Insights from Microarray Analysis","volume":"39","author":"Lee","year":"2025","journal-title":"Korean J. Ophthalmol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"14334","DOI":"10.1523\/JNEUROSCI.2770-09.2009","article-title":"Neuroprotective and axon growth-promoting effects following inflammatory stimulation on mature retinal ganglion cells in mice depend on ciliary neurotrophic factor and leukemia inhibitory factor","volume":"29","author":"Leibinger","year":"2009","journal-title":"J. Neurosci."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"815","DOI":"10.2174\/0929866529666220905105800","article-title":"The Roles of Ciliary Neurotrophic Factor\u2014From Neuronutrition to Energy Metabolism","volume":"29","author":"Guo","year":"2022","journal-title":"Protein Pept. Lett."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"e2017282118","DOI":"10.1073\/pnas.2017282118","article-title":"Chemokine CCL5 promotes robust optic nerve regeneration and mediates many of the effects of CNTF gene therapy","volume":"118","author":"Xie","year":"2021","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1656","DOI":"10.1093\/brain\/awu037","article-title":"Combined suppression of CASP2 and CASP6 protects retinal ganglion cells from apoptosis and promotes axon regeneration through CNTF-mediated JAK\/STAT signalling","volume":"137","author":"Vigneswara","year":"2014","journal-title":"Brain"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"3896","DOI":"10.1073\/pnas.0600236103","article-title":"Ciliary neurotrophic factor (CNTF) for human retinal degeneration: Phase I trial of CNTF delivered by encapsulated cell intraocular implants","volume":"103","author":"Sieving","year":"2006","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Tsung, T.-H., Tsai, Y.-C., Lee, H.-P., Chen, Y.-H., and Lu, D.-W. (2023). Biodegradable Polymer-Based Drug-Delivery Systems for Ocular Diseases. Int. J. Mol. Sci., 24.","DOI":"10.3390\/ijms241612976"},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Burhan, A.M., Klahan, B., Cummins, W., Andr\u00e9s-Guerrero, V., Byrne, M.E., O\u2019Reilly, N.J., Chauhan, A., Fitzhenry, L., and Hughes, H. (2021). Posterior Segment Ophthalmic Drug Delivery: Role of Muco-Adhesion with a Special Focus on Chitosan. Pharmaceutics, 13.","DOI":"10.3390\/pharmaceutics13101685"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"498","DOI":"10.1177\/08853282241277345","article-title":"Injectable nanogels to improve triamcinolone acetonide delivery and toxicity on the treatment of eye diseases","volume":"39","author":"Erdal","year":"2025","journal-title":"J. Biomater. Appl."},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Padavannil, A., Ayala-Hernandez, M.G., Castellanos-Silva, E.A., and Letts, J.A. (2022). The Mysterious Multitude: Structural Perspective on the Accessory Subunits of Respiratory Complex I. Front. Mol. Biosci., 8.","DOI":"10.3389\/fmolb.2021.798353"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"1252","DOI":"10.1016\/j.freeradbiomed.2012.07.021","article-title":"Cytochrome c oxidase dysfunction in oxidative stress","volume":"53","author":"Srinivasan","year":"2012","journal-title":"Free Radic. Biol. Med."},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Kozhukhar, N., and Alexeyev, M.F. (2023). 35 Years of TFAM Research: Old Protein, New Puzzles. Biology, 12.","DOI":"10.3390\/biology12060823"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"e1839","DOI":"10.1038\/cddis.2015.180","article-title":"DRP1 inhibition rescues retinal ganglion cells and their axons by preserving mitochondrial integrity in a mouse model of glaucoma","volume":"6","author":"Kim","year":"2015","journal-title":"Cell Death Dis."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"4","DOI":"10.1167\/iovs.63.9.4","article-title":"Ciliary Neurotrophic Factor Derived from Astrocytes Protects Retinal Ganglion Cells Through PI3K\/AKT, JAK\/STAT, and MAPK\/ERK Pathways","volume":"63","author":"Lee","year":"2022","journal-title":"Investig. Ophthalmol. Vis. Sci."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"485","DOI":"10.1080\/24701394.2017.1307976","article-title":"Genetic diversities of MT-ND1 and MT-ND2 genes are associated with high-altitude adaptation in yak","volume":"29","author":"Shi","year":"2017","journal-title":"Mitochondrial DNA Part A DNA Mapp. Seq. Anal."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"S213","DOI":"10.33549\/physiolres.930000.53.S213","article-title":"Genetic defects of cytochrome c oxidase assembly","volume":"53","author":"Pecina","year":"2004","journal-title":"Physiol. Res."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"4381","DOI":"10.1007\/s12035-023-03841-7","article-title":"The Protective Mechanism of TFAM on Mitochondrial DNA and its Role in Neurodegenerative Diseases","volume":"61","author":"Song","year":"2023","journal-title":"Mol. Neurobiol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"151","DOI":"10.1002\/ana.410350206","article-title":"Ciliary neurotrophic factor: Pharmacokinetics and acute-phase response in rat","volume":"35","author":"Dittrich","year":"1994","journal-title":"Ann. Neurol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"122486","DOI":"10.1016\/j.carbpol.2024.122486","article-title":"Absorption enhancement strategies in chitosan-based nanosystems and hydrogels intended for ocular delivery: Latest advances for optimization of drug permeation","volume":"343","author":"Dmour","year":"2024","journal-title":"Carbohydr. Polym."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"106575","DOI":"10.1016\/j.reactfunctpolym.2025.106575","article-title":"Chitosan-based delivery systems for ocular therapeutics: Advances, challenges, and future perspectives","volume":"219","author":"Sau","year":"2026","journal-title":"React. Funct. Polym."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"419","DOI":"10.1016\/j.jconrel.2020.07.031","article-title":"Intravitreal hydrogels for sustained release of therapeutic proteins","volume":"326","author":"Ilochonwu","year":"2020","journal-title":"J. Control. Release"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"207","DOI":"10.1007\/s10633-018-9638-x","article-title":"ISCEV extended protocol for the photopic negative response (PhNR) of the full-field electroretinogram","volume":"136","author":"Frishman","year":"2018","journal-title":"Doc. Ophthalmol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"1879","DOI":"10.1167\/iovs.18-25272","article-title":"The Relationship Between the Photopic Negative Response and Retinal Ganglion Cell Topography","volume":"60","author":"Morny","year":"2019","journal-title":"Investig. Ophthalmol. Vis. Sci."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"233","DOI":"10.1080\/1086508X.2008.11079688","article-title":"Visual Evoked Potentials","volume":"48","author":"Nehamkin","year":"2008","journal-title":"Am. J. Electroneurodiagnostic Technol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"10806","DOI":"10.1038\/s41467-024-54982-9","article-title":"DLK-dependent axonal mitochondrial fission drives degeneration after axotomy","volume":"15","author":"Nebiyou","year":"2024","journal-title":"Nat. Commun."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"4","DOI":"10.32607\/actanaturae.11610","article-title":"Fifty Years of Research on Protonophores: Mitochondrial Uncoupling as a Basis for Therapeutic Action","volume":"14","author":"Kotova","year":"2022","journal-title":"Acta Naturae"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"81","DOI":"10.1038\/s41392-022-01289-0","article-title":"Regeneration and functional recovery of the completely transected optic nerve in adult rats by CNTF-chitosan","volume":"8","author":"Liu","year":"2023","journal-title":"Signal Transduct. Target. Ther."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"201","DOI":"10.1006\/ncmn.1993.1025","article-title":"Purification of Rat Retinal Ganglion Cells by Immunopanning","volume":"2","author":"Barres","year":"1993","journal-title":"Neuroprotocols"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/27\/16\/7200\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,8,12]],"date-time":"2026-08-12T11:04:03Z","timestamp":1786532643000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/27\/16\/7200"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,8,12]]},"references-count":34,"journal-issue":{"issue":"16","published-online":{"date-parts":[[2026,8]]}},"alternative-id":["ijms27167200"],"URL":"https:\/\/doi.org\/10.3390\/ijms27167200","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,8,12]]}}}