{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,14]],"date-time":"2026-07-14T18:26:03Z","timestamp":1784053563369,"version":"3.55.0"},"reference-count":71,"publisher":"Springer Science and Business Media LLC","issue":"6","license":[{"start":{"date-parts":[[2025,4,11]],"date-time":"2025-04-11T00:00:00Z","timestamp":1744329600000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2025,4,11]],"date-time":"2025-04-11T00:00:00Z","timestamp":1744329600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"funder":[{"DOI":"10.13039\/501100015068","name":"Universidade de Santiago de Compostela","doi-asserted-by":"crossref","id":[{"id":"10.13039\/501100015068","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Drug Deliv. and Transl. Res."],"published-print":{"date-parts":[[2026,6]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Uveal melanoma is the most common primary intraocular tumor in adults that appears mainly in the choroid, ciliary body, or iris. Standard non-surgical treatment consists of radiotherapy and chemotherapy by intravitreal injection of drugs, which is hampered by unpleasant side effects and high recurrence rate. As an alternative strategy, gene therapy has the capacity of targeting specific pathways in cancer cells by introducing tumor suppressor sequences that are rendered therapeutically effective by the use of suitable delivery vectors. In this paper, we describe the development of protamine nanocapsules as potential gene delivery carriers to the eye by its topical administration. These nanocapsules, composed of spherical oily nano-droplets surrounded by a protamine shell with small particle size (\u2264\u2009250\u00a0nm) and positive surface charge (+\u200933 mV), can efficiently associate different nucleic acids such as pDNA and miRNA, and release them in biorelevant media. In vitro studies evidenced their low cytotoxicity and efficient internalization with 36% of TAMRA-positive cells, and transfection capacity from doses of 0.5\u00a0\u00b5g of pDNA in uveal melanoma cells. Finally, the results also confirmed their efficient interaction and permeation in primary patient-derived 3D corneal models with a reduction of up to 60% in TEER values, without altering the structure of this ocular barrier.<\/jats:p>\n                  <jats:p>\n                    <jats:bold>Graphical abstract<\/jats:bold>\n                  <\/jats:p>","DOI":"10.1007\/s13346-025-01849-1","type":"journal-article","created":{"date-parts":[[2025,4,11]],"date-time":"2025-04-11T07:59:43Z","timestamp":1744358383000},"page":"1754-1772","update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Protamine nanocapsules as gene delivery carriers for the treatment of intraocular tumors"],"prefix":"10.1007","volume":"16","author":[{"given":"Sheila","family":"Barrios-Esteban","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ignacio","family":"Alcalde","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Manuel","family":"Chac\u00f3n","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jes\u00fas","family":"Merayo-Lloves","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mar\u00eda","family":"de la Fuente","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6187-7717","authenticated-orcid":false,"given":"Noemi","family":"Csaba","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2025,4,11]]},"reference":[{"key":"1849_CR1","doi-asserted-by":"publisher","first-page":"677","DOI":"10.1007\/s10555-018-9762-9","volume":"37","author":"A Maheshwari","year":"2018","unstructured":"Maheshwari A, Finger PT. Cancers of the eye. Cancer Metastasis Rev. 2018;37:677\u201390. https:\/\/doi.org\/10.1007\/s10555-018-9762-9.","journal-title":"Cancer Metastasis Rev"},{"key":"1849_CR2","doi-asserted-by":"publisher","first-page":"531","DOI":"10.1016\/j.mcna.2021.02.005","volume":"105","author":"BK Williams","year":"2021","unstructured":"Williams BK, di Nicola M. Ocular oncology-primary and metastatic malignances. Med Clin. 2021;105:531\u201350. https:\/\/doi.org\/10.1016\/j.mcna.2021.02.005.","journal-title":"Med Clin"},{"key":"1849_CR3","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1038\/s41572-020-0158-0","volume":"24","author":"MJ Jager","year":"2020","unstructured":"Jager MJ, Shields CL, Cebulla CM, Abdel-Rahman MH, Grossniklaus HE, Stern MH, Carvajal RD, Belfort RN, Jia R, Shields JA, Damato BE. Uveal melanoma. Nat Rev Dis Primers. 2020;24:1\u201325. https:\/\/doi.org\/10.1038\/s41572-020-0158-0.","journal-title":"Nat Rev Dis Primers"},{"key":"1849_CR4","doi-asserted-by":"publisher","first-page":"1","DOI":"10.3390\/cancers13081933","volume":"13","author":"G Giannaccare","year":"2021","unstructured":"Giannaccare G, Bernabei F, Angi M, Pellegrini M, Maestri A, Romano V, Scorcia V, Rothschild PR. Iatrogenic ocular surface diseases occurring during and\/or after different treatments for ocular tumours. Cancers. 2021;13:1\u201317. https:\/\/doi.org\/10.3390\/cancers13081933.","journal-title":"Cancers"},{"key":"1849_CR5","doi-asserted-by":"publisher","first-page":"1","DOI":"10.31083\/j.fbl2702072","volume":"27","author":"S Ka\u0161telan","year":"2020","unstructured":"Ka\u0161telan S, Mrazovac DZ, Ivankovi\u0107 M, Markovi\u0107 I, Gverovi\u0107 AA. Liver metastasis in uveal melanoma-treatment options and clinical outcome. Front Biosci. 2020;27:1\u201316. https:\/\/doi.org\/10.31083\/j.fbl2702072.","journal-title":"Front Biosci"},{"key":"1849_CR6","doi-asserted-by":"publisher","first-page":"615","DOI":"10.17219\/acem\/146581","volume":"31","author":"MS Nowak","year":"2022","unstructured":"Nowak MS, Romanowska-Dixon B, Grabska-Liberek I, \u017burek M. Incidence, and survival of ocular melanoma in National cancer registry of Poland in 2010\u20132017. Adv Clin Exp Med. 2022;31:615\u201321. https:\/\/doi.org\/10.17219\/acem\/146581.","journal-title":"Adv Clin Exp Med"},{"key":"1849_CR7","doi-asserted-by":"publisher","first-page":"1","DOI":"10.3892\/ijo.2021.5190","volume":"58","author":"M Fallico","year":"2021","unstructured":"Fallico M, Raciti G, Longo A, Reibaldi M, Bonfiglio V, Russo A, Caltabiano R, Gattuso G, Falzone L, Avitabile T. Current molecular and clinical insights into uveal melanoma. Int J Oncol. 2021;58:1\u201322. https:\/\/doi.org\/10.3892\/ijo.2021.5190.","journal-title":"Int J Oncol"},{"key":"1849_CR8","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1002\/advs.202003699","volume":"9","author":"Z Tang","year":"2022","unstructured":"Tang Z, Fan X, Chen Y, Gu P. Ocular nanomedicine. Adv Sci. 2022;9:1\u201336. https:\/\/doi.org\/10.1002\/advs.202003699.","journal-title":"Adv Sci"},{"key":"1849_CR9","doi-asserted-by":"publisher","first-page":"4171","DOI":"10.2147\/IJGM.S299144","volume":"14","author":"S Valasapalli","year":"2021","unstructured":"Valasapalli S, Guddati AK. Nation-wide trends in incidence-based mortality of patients with ocular melanoma in USA: 200 to 2018. Int J Gen Med. 2021;14:4171\u20136. https:\/\/doi.org\/10.2147\/IJGM.S299144.","journal-title":"Int J Gen Med"},{"key":"1849_CR10","doi-asserted-by":"publisher","first-page":"3805","DOI":"10.2147\/IJN.S48950","volume":"8","author":"Y Wang","year":"2013","unstructured":"Wang Y, Mo L, Wei W, Shi X. Efficacy, and safety of dendrimer nanoparticles with coexpression of tumor necrosis factor-\u03b1 and herpes simplex virus thymidine kinase in gene radiotherapy of the human uveal melanoma OCM-1 cell line. Int J Nanomed. 2013;8:3805\u201316. https:\/\/doi.org\/10.2147\/IJN.S48950.","journal-title":"Int J Nanomed"},{"key":"1849_CR11","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1016\/j.ijpharm.2019.118894","volume":"573","author":"L Xie","year":"2020","unstructured":"Xie L, Yang Y, Shen J. Efficient Inhibition of uveal melanoma via ternary SiRNA complexes. Int J Pharm. 2020;573:1\u201311. https:\/\/doi.org\/10.1016\/j.ijpharm.2019.118894.","journal-title":"Int J Pharm"},{"key":"1849_CR12","doi-asserted-by":"publisher","first-page":"1","DOI":"10.3390\/cancers13205192","volume":"13","author":"C Yang","year":"2021","unstructured":"Yang C, Wang R, Hardy P. Potential of miRNA-based nanotherapeutics for uveal melanoma. Cancers. 2021;13:1\u201317. https:\/\/doi.org\/10.3390\/cancers13205192.","journal-title":"Cancers"},{"key":"1849_CR13","doi-asserted-by":"publisher","first-page":"243","DOI":"10.56933\/Pharmaspire.2023.15137","volume":"15","author":"A Verma","year":"2023","unstructured":"Verma A, Awasthi A. Developing non-viral or viral vectors for efficient and targeted delivery of genetic material, such as DNA or RNA, for gene therapy applications. Pharmaspire. 2023;15:243\u201356. https:\/\/doi.org\/10.56933\/Pharmaspire.2023.15137.","journal-title":"Pharmaspire"},{"key":"1849_CR14","unstructured":"Diebold Y, Enr\u00edquez de Salamanca A, Calonge M, Vila A, Carvalho ELS, de la Fuente M, Seijo B, Alonso MJ. Ocular surface in vivo tolerance to new nanoparticle polymer systems designed for drug delivery. Invest Ophthalmol Vis Sci 2005; 46."},{"key":"1849_CR15","doi-asserted-by":"crossref","unstructured":"Diebold Y, Enr\u00edquez de Salamanca A, Orea M, S\u00e1ez V, Garc\u00eda-V\u00e1zquez C, de la Fuenta M, Seijo B, Alonso MJ, Calonge M. In vivo uptake of nanoparticulate drug delivery systems by ocular surface structures. Invest Ophthalmol Vis Sci 2006; 47.","DOI":"10.1167\/iovs.05-0495"},{"key":"1849_CR16","doi-asserted-by":"crossref","unstructured":"Contreras L, L\u00f3pez-Garc\u00eda A, de la Fuente M, Seijo B, Alonso MJ, Diebold Y. Hyaluronic receptor-mediated uptake of hyaluronic acid-chitosan nanoparticles in ocular epithelial cells. Invest Ophthalmol Vis Sci 2008; 49.","DOI":"10.1167\/iovs.07-1077"},{"key":"1849_CR17","doi-asserted-by":"publisher","first-page":"550","DOI":"10.1097\/ICO.0b013e3181bd9eee","volume":"29","author":"L Contreras-Ruiz","year":"2010","unstructured":"Contreras-Ruiz L, de la Fuente M, Garc\u00eda-V\u00e1zquez C, S\u00e1ez V, Seijo B, Alonso MJ, Calonge M, Diebold Y. Ocular tolerance to a topical formulation of hyaluronic acid and chitosan-based nanoparticles. Cornea. 2010;29:550\u20138. https:\/\/doi.org\/10.1097\/ICO.0b013e3181bd9eee.","journal-title":"Cornea"},{"key":"1849_CR18","doi-asserted-by":"publisher","first-page":"708","DOI":"10.1007\/s13346-016-0312-0","volume":"6","author":"S Reimondez-Troiti\u00f1o","year":"2016","unstructured":"Reimondez-Troiti\u00f1o S, Alcalde I, Csaba N, \u00cd\u00f1igo-Portugu\u00e9s A, de la Fuente M, Bech F, Riestra AC, Merayo-Lloves J, Alonso MJ. Polymeric nanocapsules: a potential new therapy for corneal wound healing. Drug Deliv Transl Res. 2016;6:708\u201321. https:\/\/doi.org\/10.1007\/s13346-016-0312-0.","journal-title":"Drug Deliv Transl Res"},{"key":"1849_CR19","doi-asserted-by":"publisher","first-page":"668","DOI":"10.1038\/gt.2008.16","volume":"15","author":"M de la Fuente","year":"2008","unstructured":"de la Fuente M, Seijo B, Alonso MJ. Bioadhesive hyaluronic-chitosan nanoparticles can transport genes across the ocular mucosa and transfect ocular tissue. Gene Ther. 2008;15:668\u201376. https:\/\/doi.org\/10.1038\/gt.2008.16.","journal-title":"Gene Ther"},{"key":"1849_CR20","doi-asserted-by":"crossref","unstructured":"Diebold Y, Contreras-Ruiz L, L\u00f3pez-Garc\u00eda A, Calonge M, de la Fuente M, Seijo B, Alonso MJ. Intracellular trafficking of hyaluronic acid-chitosan nanoparticles in ocular epithelial cells. Invest Ophthalmol Vis Sci 2008; 49.","DOI":"10.1167\/iovs.07-1077"},{"key":"1849_CR21","doi-asserted-by":"publisher","first-page":"2016","DOI":"10.1167\/iovs.07-1077","volume":"49","author":"M de la Fuente","year":"2008","unstructured":"de la Fuente M, Seijo B, Alonso MJ. Novel hyaluronic acid-chitosan nanoparticles for ocular gene therapy. Physiol Pharmacol. 2008;49:2016\u201324. https:\/\/doi.org\/10.1167\/iovs.07-1077.","journal-title":"Physiol Pharmacol"},{"key":"1849_CR22","doi-asserted-by":"publisher","first-page":"70","DOI":"10.1002\/jgm.1125","volume":"10","author":"M Hornof","year":"2007","unstructured":"Hornof M, de la Fuente M, Hallikainen M, Tammi RH, Urtti A. Low molecular weight hyaluronan shielding of DNA\/PEI polyplexes facilitates CD44 receptor mediated uptake in human corneal epithelial cells. J Gene Med. 2007;10:70\u201380. https:\/\/doi.org\/10.1002\/jgm.1125.","journal-title":"J Gene Med"},{"key":"1849_CR23","first-page":"279","volume":"17","author":"L Contreras-Ruiz","year":"2011","unstructured":"Contreras-Ruiz L, de la Fuente M, P\u00e1rraga JE, L\u00f3pez-Garc\u00eda A, Fern\u00e1ndez I, Seijo B, S\u00e1nchez A, Calonge M, Diebold Y. Intracellular trafficking of hyaluronic acid-chitosan oligomer-based nanoparticles in cultured human ocular surface cells. Mol Vis. 2011;17:279\u201390.","journal-title":"Mol Vis"},{"key":"1849_CR24","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1016\/j.ajo.2018.11.007","volume":"199","author":"N Venkateswaran","year":"2018","unstructured":"Venkateswaran N, Mercado C, Galor A, Karp CL. Comparison of topical 5-fluorouracil and interferon alfa-2b as primary treatment modalities for ocular surface squamous neoplasia. Am J Ophtalmol. 2018;199:1\u201310. https:\/\/doi.org\/10.1016\/j.ajo.2018.11.007.","journal-title":"Am J Ophtalmol"},{"key":"1849_CR25","doi-asserted-by":"publisher","first-page":"1","DOI":"10.3390\/ijms241612976","volume":"24","author":"TH Tsung","year":"2023","unstructured":"Tsung TH, Tsai YC, Lee HP, Chen YH, Lu DW. Biodegradable polymer-based drug-delivery systems for ocular diseases. Int J Mol Sci. 2023;24:1\u201332. https:\/\/doi.org\/10.3390\/ijms241612976.","journal-title":"Int J Mol Sci"},{"key":"1849_CR26","doi-asserted-by":"publisher","first-page":"1","DOI":"10.3390\/pharmaceutics14040799","volume":"14","author":"R Varela-Fern\u00e1ndez","year":"2022","unstructured":"Varela-Fern\u00e1ndez R, Garc\u00eda-Otero Xurxo, D\u00edaz-Tom\u00e9 V, Regueiro U, L\u00f3pez-L\u00f3pez M, Gonz\u00e1lez-Barcia M, Lema MI, Otero-Espinar FJ. Mucoadhesive PLGA nanospheres and nanocapsules for lactoferrin controlled ocular delivery. Pharm. 2022;14:1\u201325. https:\/\/doi.org\/10.3390\/pharmaceutics14040799.","journal-title":"Pharm"},{"key":"1849_CR27","doi-asserted-by":"publisher","first-page":"278","DOI":"10.18433\/J3DP43","volume":"17","author":"H Almeida","year":"2014","unstructured":"Almeida H, Amaral MH, Lob\u00e3o P, Silva AC, Loboa JM. Applications of polymeric and lipid nanoparticles in ophthalmic pharmaceutical formulations: present and future considerations. J Pharm Pharm Sci. 2014;17:278\u201393. https:\/\/doi.org\/10.18433\/J3DP43.","journal-title":"J Pharm Pharm Sci"},{"key":"1849_CR28","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1016\/j.jddst.2019.101389","volume":"55","author":"R Suri","year":"2020","unstructured":"Suri R, Beg S, Kohli K. Target strategies for drug delivery bypassing ocular barriers. J Drug Deliv Sci Technol. 2020;55:1\u201362. https:\/\/doi.org\/10.1016\/j.jddst.2019.101389.","journal-title":"J Drug Deliv Sci Technol"},{"key":"1849_CR29","doi-asserted-by":"publisher","first-page":"71","DOI":"10.1016\/j.jconrel.2021.06.011","volume":"336","author":"RS Dave","year":"2021","unstructured":"Dave RS, Goostrey TC, Ziolkowska M, Czerny-Holownia S, Hoare T, Sheardown H. Ocular drug delivery to the anterior segment using nanocarriers: a mucoadhesive\/mucopenetrative perspective. JCR. 2021;336:71\u201388. https:\/\/doi.org\/10.1016\/j.jconrel.2021.06.011.","journal-title":"JCR"},{"key":"1849_CR30","doi-asserted-by":"publisher","unstructured":"Silva B, Braz BS, Delgado E, Goncalves L. Colloidal nanosystems with mucoadhesive properties designed for ocular topical delivery. 2021. Inter. J. Pharm. 2021;606:1\u201326. https:\/\/doi.org\/10.1016\/j.ijpharm.2021.120873","DOI":"10.1016\/j.ijpharm.2021.120873"},{"key":"1849_CR31","doi-asserted-by":"publisher","first-page":"1","DOI":"10.3390\/ijms222212368","volume":"22","author":"A Vaneev","year":"2021","unstructured":"Vaneev A, Tikhomirova V, Chesnokova N, Popoya E, Beznos O, Kost O, Klyacho N. Nanotechnology for topical drug delivery to the anterior segment of the eye. Int J Mol Sci. 2021;22:1\u201330. https:\/\/doi.org\/10.3390\/ijms222212368.","journal-title":"Int J Mol Sci"},{"key":"1849_CR32","doi-asserted-by":"publisher","first-page":"449","DOI":"10.1016\/j.jddst.2018.04.007","volume":"45","author":"S J\u00f3hannsd\u00f3ttir","year":"2018","unstructured":"J\u00f3hannsd\u00f3ttir S, Jansook P, Stef\u00e1nsson E, Kristinsd\u00f3ttir \u00cdM, F\u00fcl\u00f6p Z, Asgr\u00edmsd\u00f3ttir GM, Thorsteindsottir M, Eir\u00edksson FF, Loftsson T. Topical drug delivery to the posterior segment of the eye: dexamethasone concentrations in various eye tissues after topical administration for up 15 days to rabbits. J Drug Deliv Sci Technol. 2018;45:449\u201354. https:\/\/doi.org\/10.1016\/j.jddst.2018.04.007.","journal-title":"J Drug Deliv Sci Technol"},{"key":"1849_CR33","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1016\/j.jconrel.2020.01.057","volume":"321","author":"C Jumelle","year":"2020","unstructured":"Jumelle C, Gholizadeh S, Annabi N, Dana R. Advances and limitations of drug delivery systems formulated as eye drops. J Control Release. 2020;321:1\u201322. https:\/\/doi.org\/10.1016\/j.jconrel.2020.01.057.","journal-title":"J Control Release"},{"key":"1849_CR34","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1016\/j.jddst.2019.101389","volume":"55","author":"R Suri","year":"2020","unstructured":"Suri R, Beg S, Kohli K. Target strategies for drug delivery by passing ocular barriers. J Drug Deliv Sci Technol. 2020;55:1\u201317. https:\/\/doi.org\/10.1016\/j.jddst.2019.101389.","journal-title":"J Drug Deliv Sci Technol"},{"key":"1849_CR35","doi-asserted-by":"publisher","first-page":"1","DOI":"10.3390\/ijms19092830","volume":"19","author":"CH Tsai","year":"2018","unstructured":"Tsai CH, Wang PY, Lin IC, Huang H, Liu GS, Tseng CL. Ocular drug delivery: role of degradable polymeric nanocarriers for ophthalmic application. Int J Mol Sci. 2018;19:1\u201320. https:\/\/doi.org\/10.3390\/ijms19092830.","journal-title":"Int J Mol Sci"},{"key":"1849_CR36","doi-asserted-by":"publisher","first-page":"449","DOI":"10.1016\/j.ejpb.2019.07.016","volume":"142","author":"S Reimondez-Troiti\u00f1o","year":"2019","unstructured":"Reimondez-Troiti\u00f1o S, Gonz\u00e1lez-Aramundiz JV, Ruiz-Ba\u00f1obre J, L\u00f3pez-L\u00f3pez R, Alonso MJ, Csaba N, de la Fuente M. Versatile Protamine nanocapsules to restore miR-145 levels and interfere tumor growth in colorectal cancer cells. Eur J Pharm Biopharm. 2019;142:449\u201359. https:\/\/doi.org\/10.1016\/j.ejpb.2019.07.016.","journal-title":"Eur J Pharm Biopharm"},{"key":"1849_CR37","doi-asserted-by":"publisher","first-page":"157","DOI":"10.1016\/j.jconrel.2018.10.022","volume":"291","author":"LN Thwala","year":"2018","unstructured":"Thwala LN, Delgado DP, Leone K, Marigo I, Benetti F, Chenlo M, Alvarez CV, Tovar S, Dieguez C, Csaba NS, Alonso MJ. Protamine nanocapsules as carriers for oral peptide delivery. J Control Release. 2018;291:157\u201368. https:\/\/doi.org\/10.1016\/j.jconrel.2018.10.022.","journal-title":"J Control Release"},{"key":"1849_CR38","doi-asserted-by":"publisher","first-page":"780","DOI":"10.3390\/cancers12040780","volume":"12","author":"Z Zeng","year":"2020","unstructured":"Zeng Z, Tung CH, Zu Y. Aptamer-equipped Protamine nanomedicine for precision lymphoma therapy. Cancers. 2020;12:780. https:\/\/doi.org\/10.3390\/cancers12040780.","journal-title":"Cancers"},{"key":"1849_CR39","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1016\/j.bioelechem.2022.108129","volume":"146","author":"M Chac\u00f3n","year":"2022","unstructured":"Chac\u00f3n M, S\u00e1nchez M, V\u00e1zquez N, Persinal-Medina M, Alonso-Alonso S, Baamonde B, Alfonso JF, Fern\u00e1ndez-Vega-Cueto L, Merayo-Lloves J, Meana \u00c1. Impedance-based non-invasive assay for ocular damage prediction on in vitro 3D reconstructed human corneal epithelium. Bioelectrochemistry. 2022;146:1\u20137. https:\/\/doi.org\/10.1016\/j.bioelechem.2022.108129.","journal-title":"Bioelectrochemistry"},{"key":"1849_CR40","doi-asserted-by":"publisher","first-page":"630","DOI":"10.1080\/10837450.2018.1550789","volume":"24","author":"MJ Alvarez-Figueroa","year":"2019","unstructured":"Alvarez-Figueroa MJ, Abarca-Riquelme JM, Gonz\u00e1lez-Aramundiz JV. Influence of Protamine shell on nanoemulsions as a carrier for cyclosporine-a skin delivery. Pharm Dev Technol. 2019;24:630\u20138. https:\/\/doi.org\/10.1080\/10837450.2018.1550789.","journal-title":"Pharm Dev Technol"},{"key":"1849_CR41","doi-asserted-by":"publisher","first-page":"9184","DOI":"10.1039\/C3CS60278H","volume":"42","author":"SM Bromfield","year":"2013","unstructured":"Bromfield SM, Wilde E, Smith DK. Heparin sensing and binding-taking supramolecular chemistry towards clinical applications. Chem Soc Rev. 2013;42:9184\u201395. https:\/\/doi.org\/10.1039\/C3CS60278H.","journal-title":"Chem Soc Rev"},{"key":"1849_CR42","doi-asserted-by":"publisher","first-page":"311","DOI":"10.1023\/A:1016015803611","volume":"13","author":"P Calvo","year":"1996","unstructured":"Calvo P, Sanchez A, Martinez J, Lopez MI, Calonge M, Pastor JC, Alonso MJ. Polyester nanocapsules as new topical ocular delivery systems for cyclosporin A. Pharm Res. 1996;13:311\u201315. https:\/\/doi.org\/10.1023\/A:1016015803611.","journal-title":"Pharm Res"},{"key":"1849_CR43","doi-asserted-by":"publisher","first-page":"105390","DOI":"10.1016\/j.tiv.2022.105390","volume":"82","author":"M Chac\u00f3n","year":"2022","unstructured":"Chac\u00f3n M, V\u00e1zquez N, Persinal-Medina M, Alonso-Alonso S, Alcalde I, Merayo-Lloves J, Meana \u00c1. In-house performance assessment of 3D QobuR-reconstructed human cornea-like epithelium (RhCE) for the evaluation of eye hazard. Toxicol Vitro. 2022;82:105390. https:\/\/doi.org\/10.1016\/j.tiv.2022.105390.","journal-title":"Toxicol Vitro"},{"key":"1849_CR44","doi-asserted-by":"publisher","first-page":"62","DOI":"10.1016\/j.jconrel.2016.11.012","volume":"245","author":"JV Gonz\u00e1lez-Aramundiz","year":"2017","unstructured":"Gonz\u00e1lez-Aramundiz JV, Presas E, Dalmau-Mena I, Mart\u00ednez-Pulgar\u00edn S, Alonso C, Escribano JM, Alonso MJ, Csaba NS. Rational design of Protamine nanocapsules as antigen delivery carriers. J Control Release. 2017;245:62\u20139. https:\/\/doi.org\/10.1016\/j.jconrel.2016.11.012.","journal-title":"J Control Release"},{"key":"1849_CR45","doi-asserted-by":"publisher","first-page":"419","DOI":"10.1016\/j.ijpharm.2011.12.016","volume":"423","author":"N Khayata","year":"2012","unstructured":"Khayata N, Abdelwahed W, Chehna MF, Charcosset C, Fessi H. Preparation of vitamin E loaded nanocapsules by the nanoprecipitation method: from laboratory scale to large scale using a membrane contactor. Int J Pharm. 2012;423:419\u201327. https:\/\/doi.org\/10.1016\/j.ijpharm.2011.12.016.","journal-title":"Int J Pharm"},{"key":"1849_CR46","doi-asserted-by":"publisher","first-page":"1","DOI":"10.3390\/pharmaceutics11050246","volume":"11","author":"JI Bussio","year":"2019","unstructured":"Bussio JI, Molina-Perea C, Gonz\u00e1lez-Aramundiz JV. Hyaluronic acid nanocapsules as a platform for Needle-Free vaccination. Pharm. 2019;11:1\u201314. https:\/\/doi.org\/10.3390\/pharmaceutics11050246.","journal-title":"Pharm"},{"key":"1849_CR47","doi-asserted-by":"publisher","first-page":"611","DOI":"10.1016\/j.xphs.2016.09.029","volume":"106","author":"G Lollo","year":"2017","unstructured":"Lollo G, Gonzalez-Paredes A, Garcia-Fuentes M, Calvo P, Torres Dolores, Alonso MF. Polyarginine nanocapsules as a potential oral peptide delivery carrier. J Pharm Sci. 2017;106:611\u20138. https:\/\/doi.org\/10.1016\/j.xphs.2016.09.029.","journal-title":"J Pharm Sci"},{"key":"1849_CR48","doi-asserted-by":"publisher","first-page":"1","DOI":"10.3390\/pharmaceutics13091377","volume":"13","author":"E Fern\u00e1ndez-Paz","year":"2021","unstructured":"Fern\u00e1ndez-Paz E, Feijoo-Siota L, Gaspar MM, Csaba N, Remu\u00f1an-L\u00f3pez C. Microencapsulated Chitosan-Based nanocapsules: A new platform for pulmonary gene delivery. Pharm. 2021;13:1\u201324. https:\/\/doi.org\/10.3390\/pharmaceutics13091377.","journal-title":"Pharm"},{"key":"1849_CR49","doi-asserted-by":"publisher","first-page":"156","DOI":"10.3390\/biomedicines10010156","volume":"10","author":"M Radanova","year":"2022","unstructured":"Radanova M, Levkova M, Mihaylova G, Manev R, Maneva M, Hadgiev R, Conev N, Donev I. Single nucleotide polymorphisms in MicroRNA genes and colorectal cancer risk and prognosis. Biomedicines. 2022;10:156. https:\/\/doi.org\/10.3390\/biomedicines10010156.","journal-title":"Biomedicines"},{"key":"1849_CR50","doi-asserted-by":"publisher","first-page":"9368","DOI":"10.3390\/ijms16059368","volume":"16","author":"J Zaloga","year":"2015","unstructured":"Zaloga J, Janko C, Agarwal R, Nowak J, M\u00fcller R, Boccaccini AR, Lee G, Odenbach S, Lyer S, Alexiou C. Different storage conditions influence biocompatibility and physicochemical properties of iron oxide nanoparticles. Int J Mol Sci. 2015;16:9368\u201384. https:\/\/doi.org\/10.3390\/ijms16059368.","journal-title":"Int J Mol Sci"},{"key":"1849_CR51","doi-asserted-by":"publisher","first-page":"178","DOI":"10.1016\/j.ijpharm.2017.06.077","volume":"529","author":"AO Elzoghby","year":"2017","unstructured":"Elzoghby AO, Mostafa SK, Helmy MW, ElDemellawy MA, Sheweita SA. Superiority of aromatase inhibitor and cyclooxygenase-2 inhibitor combined delivery: Hyaluronate-targeted versus pegylated Protamine nanocapsules for breast cancer therapy. Int J Pharm. 2017;529:178\u201392. https:\/\/doi.org\/10.1016\/j.ijpharm.2017.06.077.","journal-title":"Int J Pharm"},{"key":"1849_CR52","doi-asserted-by":"publisher","first-page":"1553","DOI":"10.1016\/j.biomaterials.2006.11.028","volume":"28","author":"Y Diebold","year":"2007","unstructured":"Diebold Y, Jarr\u00edn M, S\u00e1ez V, Carvalho EL, Orea M, Calonge M, Seijo B, Alonso MJ. Ocular drug delivery by liposome\u2013chitosan nanoparticle complexes (LCS-NP). Biomater. 2007;28:1553\u201364. https:\/\/doi.org\/10.1016\/j.biomaterials.2006.11.028.","journal-title":"Biomater"},{"key":"1849_CR53","doi-asserted-by":"publisher","first-page":"109","DOI":"10.1016\/j.jconrel.2016.10.002","volume":"243","author":"LN Thwala","year":"2016","unstructured":"Thwala LN, Beloqui A, Csaba NS, Gonz\u00e1lez-Touceda D, Tovar S, Dieguez C, Alonso MJ, Pr\u00e9at V. The interaction of Protamine nanocapsules with the intestinal epithelium: A mechanistic approach. J Control Release. 2016;243:109\u201320. https:\/\/doi.org\/10.1016\/j.jconrel.2016.10.002.","journal-title":"J Control Release"},{"key":"1849_CR54","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1016\/j.biopha.2019.109014","volume":"116","author":"MP Pereira","year":"2019","unstructured":"Pereira MP, de Gomes MG, Izoton JC, Nakama KA, Dos Santos RB, Pinto Savall AS, Ramalho JB, Roman SS, Luchese C, Cibin FW, Pinton S, Haas SE. Cationic and anionic unloaded polymeric nanocapsules: toxicological evaluation in rats shows low toxicity. Biomed Pharmacother. 2019;116:1\u20138. https:\/\/doi.org\/10.1016\/j.biopha.2019.109014.","journal-title":"Biomed Pharmacother"},{"key":"1849_CR55","doi-asserted-by":"publisher","first-page":"41","DOI":"10.1016\/S0378-5173(97)00083-5","volume":"153","author":"P Calvo","year":"1997","unstructured":"Calvo P, Vila-Jato JL, Alonso MJ. Evaluation of cationic polymer-coated nanocapsules as ocular drug carriers. Inter J Pharm. 1997;153:41\u201350. https:\/\/doi.org\/10.1016\/S0378-5173(97)00083-5.","journal-title":"Inter J Pharm"},{"key":"1849_CR56","doi-asserted-by":"publisher","first-page":"373","DOI":"10.1016\/j.jconrel.2023.03.002","volume":"356","author":"J Wu","year":"2023","unstructured":"Wu J, Jones N, Fayez NAL, Chao PH, Wu A, de Araujo DR, Rouhollahi E, Jia A, Li SD. Protamine-mediated efficient transcellular and transmucosal delivery of proteins. J Control Release. 2023;356:373\u201385. https:\/\/doi.org\/10.1016\/j.jconrel.2023.03.002.","journal-title":"J Control Release"},{"key":"1849_CR57","doi-asserted-by":"publisher","first-page":"305","DOI":"10.1007\/s00216-008-2231-5","volume":"392","author":"S Kalkhof","year":"2008","unstructured":"Kalkhof S, Sinz A. Chances, and pitfalls of chemical cross-linking with amine-reactive N-hydroxysuccinimide esters. Anal Bioanal Chem. 2008;392:305\u201312. https:\/\/doi.org\/10.1007\/s00216-008-2231-5.","journal-title":"Anal Bioanal Chem"},{"key":"1849_CR58","doi-asserted-by":"publisher","first-page":"961","DOI":"10.1038\/sj.gt.3300484","volume":"4","author":"FL Sorgi","year":"1997","unstructured":"Sorgi FL, Bhattacharya S, Huang L. Protamine sulfate enhaces lipid-mediated gene transfer. Gene Ther. 1997;4:961\u20138. https:\/\/doi.org\/10.1038\/sj.gt.3300484.","journal-title":"Gene Ther"},{"key":"1849_CR59","doi-asserted-by":"publisher","first-page":"877","DOI":"10.3390\/pharmaceutics13060877","volume":"13","author":"NT Jarzebska","year":"2021","unstructured":"Jarzebska NT, Mellett M, Frei J, K\u00fcndig TM, Pascolo S. Protamine-based strategies for RNA transfection. Pharmaceutics. 2021;13:877. https:\/\/doi.org\/10.3390\/pharmaceutics13060877.","journal-title":"Pharmaceutics"},{"key":"1849_CR60","doi-asserted-by":"publisher","first-page":"1240","DOI":"10.1021\/bc0501451","volume":"16","author":"F Reynolds","year":"2005","unstructured":"Reynolds F, Weissleder R, Josephson L. Protamine as an efficient membrane translocating peptide. Bioconjug Chem. 2005;16:1240\u20135. https:\/\/doi.org\/10.1021\/bc0501451.","journal-title":"Bioconjug Chem"},{"key":"1849_CR61","doi-asserted-by":"publisher","first-page":"575","DOI":"10.1002\/jbm.a.34859","volume":"102","author":"MC Shin","year":"2013","unstructured":"Shin MC, Zhang J, Min KA, Lee K, Byun Y, David AE, He H, Yang VC. Cell-penetrating peptides: achievements and challenges in application for cancer treatment. J Biomed Mater Res A. 2013;102:575\u201387. https:\/\/doi.org\/10.1002\/jbm.a.34859.","journal-title":"J Biomed Mater Res A"},{"key":"1849_CR62","doi-asserted-by":"publisher","first-page":"384","DOI":"10.1016\/j.ejpb.2010.07.012","volume":"76","author":"E Vighi","year":"2010","unstructured":"Vighi E, Montanari M, Ruozi B, Tosi G, Magli A, Leo E. Nuclear localization of cationic solid lipid nanoparticles containing Protamine as transfection promoter. Eur J Pharm Biopharm. 2010;76:384\u201393. https:\/\/doi.org\/10.1016\/j.ejpb.2010.07.012.","journal-title":"Eur J Pharm Biopharm"},{"key":"1849_CR63","doi-asserted-by":"publisher","first-page":"495","DOI":"10.1016\/j.ejpb.2011.06.005","volume":"79","author":"D Delgado","year":"2011","unstructured":"Delgado D, del Pozo-Rodr\u00edguez A, Solin\u00eds MA, Rodr\u00edguez-Gasc\u00f3n A. Understanding the mechanism of Protamine in solid lipid nanoparticle-based lipofection: the importance of the entry pathway. Eur J Pharm Biopharm. 2011;79:495\u2013502. https:\/\/doi.org\/10.1016\/j.ejpb.2011.06.005.","journal-title":"Eur J Pharm Biopharm"},{"key":"1849_CR64","doi-asserted-by":"publisher","first-page":"410","DOI":"10.1016\/j.carbpol.2017.02.065","volume":"165","author":"KY Lu","year":"2017","unstructured":"Lu KY, Li R, Hsu CH, Lin CW, Chou SC, Tsai ML, Mi FL. Development of a new type of multifunctional fucoidan-based nanoparticles for anticancer drug delivery. Carbohydr Polym. 2017;165:410\u201320. https:\/\/doi.org\/10.1016\/j.carbpol.2017.02.065.","journal-title":"Carbohydr Polym"},{"key":"1849_CR65","doi-asserted-by":"publisher","unstructured":"Shamarekh KS, Gad HA, Soliman ME, Sammour OA. Development and evaluation of protamine-coated PLGA nanoparticles for nose-to brain delivery of tacrine: in vitro and in vivo assessment. J. Drug Deliv. Sci. Technol. 2020;57:1\u201361. https:\/\/doi.org\/10.1016\/j.jddst.2020.101724","DOI":"10.1016\/j.jddst.2020.101724"},{"key":"1849_CR66","doi-asserted-by":"publisher","first-page":"1","DOI":"10.3390\/polym14122403","volume":"14","author":"I Ahmad","year":"2022","unstructured":"Ahmad I, Khan MFA, Rahdar A, Hussain S, Tareen FK, Salim MW, Ajalli N, Amirzada MI, Khan A. Design and evaluation of pH sensitive PEG-protamine nanocomplex of doxorubicin for treatment of breast cancer. Polymers. 2022;14:1\u201313. https:\/\/doi.org\/10.3390\/polym14122403.","journal-title":"Polymers"},{"key":"1849_CR67","doi-asserted-by":"publisher","first-page":"310","DOI":"10.1039\/D0MA00353K","volume":"2","author":"C Guido","year":"2021","unstructured":"Guido C, Testini M, D\u2019Amone S, Cortese B, Grano M, Gigli G, Palam\u00e0 IE. Capsid-like biodegradable poly-glycolic acid nanoparticles for a long-time release of nucleic acid molecules. Mater Adv. 2021;2:310\u201321. https:\/\/doi.org\/10.1039\/D0MA00353K.","journal-title":"Mater Adv"},{"key":"1849_CR68","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1007\/s12247-018-9341-1","volume":"14","author":"D Hernandez-Patlan","year":"2019","unstructured":"Hernandez-Patlan D, Solis-Cruz B, Cano-Vega MA, Beyssac E, Garrait G, Hernandez-Velasco X, Lopez-Arellano R, Tellez G, Rivera-Rodriguez GR. Development of Chitosan and alginate nanocapsules to increase the solubility, permeability, and stability of Curcumin. J Pharm Innov. 2019;14:1\u20139. https:\/\/doi.org\/10.1007\/s12247-018-9341-1.","journal-title":"J Pharm Innov"},{"key":"1849_CR69","doi-asserted-by":"publisher","first-page":"164","DOI":"10.1016\/j.ejpb.2019.01.023","volume":"136","author":"M Chac\u00f3n","year":"2019","unstructured":"Chac\u00f3n M, V\u00e1zquez N, Berisa S, Persinal M, S\u00e1nchez M, Baamonde B, Alfonso JF, Fern\u00e1ndez-Vega Cueto L, Merayo-Lloves J, Meana \u00c1. QobuR-a new in vitro human corneal epithelial model for preclinical drug screening. Eur J Pharm Biopharm. 2019;136:164\u201373. https:\/\/doi.org\/10.1016\/j.ejpb.2019.01.023.","journal-title":"Eur J Pharm Biopharm"},{"key":"1849_CR70","doi-asserted-by":"publisher","first-page":"132","DOI":"10.1007\/s12247-018-9341-1","volume":"14","author":"D Hernandez-Patlan","year":"2018","unstructured":"Hernandez-Patlan D, Solis-Cruz B, Cano-Vega MA, Beyssac E, Garrait G, Hernandez-Velasco X, Lopez-Arellano R, Tellez G, Rivera-Rodriguez GR. Development of Chitosan and alginate nanocapsules to increase the solubility, permeability and stability of Curcumin. J Pharm Innov. 2018;14:132\u201340. https:\/\/doi.org\/10.1007\/s12247-018-9341-1.","journal-title":"J Pharm Innov"},{"key":"1849_CR71","doi-asserted-by":"publisher","first-page":"2779","DOI":"10.1007\/s00018-012-0951-3","volume":"69","author":"L Li","year":"2012","unstructured":"Li L, Hartley R, Reiss B, Sun Y, Pu J, Wu D, Lin F, Hoang T, Yamada S, Jiang J, Zhao M. E-cadherin plays an essential role in collective directional migration of large epithelial sheets. Cell Mol Life Sci. 2012;69:2779\u201389. https:\/\/doi.org\/10.1007\/s00018-012-0951-3.","journal-title":"Cell Mol Life Sci"}],"container-title":["Drug Delivery and Translational Research"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1007\/s13346-025-01849-1.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/link.springer.com\/article\/10.1007\/s13346-025-01849-1","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1007\/s13346-025-01849-1.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,5,18]],"date-time":"2026-05-18T12:40:22Z","timestamp":1779108022000},"score":1,"resource":{"primary":{"URL":"https:\/\/link.springer.com\/10.1007\/s13346-025-01849-1"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,4,11]]},"references-count":71,"journal-issue":{"issue":"6","published-print":{"date-parts":[[2026,6]]}},"alternative-id":["1849"],"URL":"https:\/\/doi.org\/10.1007\/s13346-025-01849-1","relation":{},"ISSN":["2190-393X","2190-3948"],"issn-type":[{"value":"2190-393X","type":"print"},{"value":"2190-3948","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,4,11]]},"assertion":[{"value":"30 March 2025","order":1,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"11 April 2025","order":2,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Declarations"}},{"value":"The human corneal epithelial cells to form the QobuR-RhCE model was collected from corneal donor rims specimens after isolation of the central corneal button for transplantation by the collaboration of Dr. Ignacio Alcalde (Laboratory of Cellular Biology and Innervation of the Ocular Surface, Fundaci\u00f3n de Investigaci\u00f3n Oftalmol\u00f3gica (FIO) at the University of Oviedo). The project with number 2020.050 was approved by the Ethics Committee for Drug Research of the Principado de Asturias. All cell lines were cultured in the conditions recommended by the manufacturers and collaborators, and only used for research purposes specifically described in this research article.","order":2,"name":"Ethics","group":{"name":"EthicsHeading","label":"Ethics approval and consent to participate"}},{"value":"All the authors read and approved the final version of the manuscript.","order":3,"name":"Ethics","group":{"name":"EthicsHeading","label":"Consent to publish"}},{"value":"Not private study sponsors had any involvement in the study design, data collection, or interpretation of data presented in this manuscript.","order":4,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}},{"value":"This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution, and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third-party material in this article are included in the article\u2019s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article\u2019s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit\n                      \n                      .","order":5,"name":"Ethics","group":{"name":"EthicsHeading","label":"Open access"}}]}}