{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,11]],"date-time":"2026-10-11T02:18:42Z","timestamp":1791685122954,"version":"4.3.4"},"reference-count":44,"publisher":"MDPI AG","issue":"8","license":[{"start":{"date-parts":[[2023,7,31]],"date-time":"2023-07-31T00:00:00Z","timestamp":1690761600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Boston Keratoprosthesis Fund"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Pharmaceutics"],"abstract":"<jats:p>Purpose: This study aimed to develop a clinically feasible and practical therapy for multi-ocular protection following ocular injury by using a thermosensitive drug delivery system (DDS) for sustained delivery of TNF-\u03b1 and VEGF inhibitors to the eye. Methods: A thermosensitive, biodegradable hydrogel DDS (PLGA-PEG-PLGA triblock polymer) loaded with 0.7 mg of adalimumab and 1.4 mg of aflibercept was injected subconjunctivally into Dutch-belted pigmented rabbits after corneal alkali injury. Control rabbits received 2 mg of IgG-loaded DDS or 1.4 mg of aflibercept-loaded DDS. Animals were followed for 3 months and assessed for tolerability and prevention of corneal neovascularization (NV), improvement of corneal re-epithelialization, inhibition of retinal ganglion cell (RGC) and optic nerve axon loss, and inhibition of immune cell infiltration into the cornea. Drug-release kinetics was assessed in vivo using an aqueous humor protein analysis. Results: A single subconjunctival administration of dual anti-TNF-\u03b1\/anti-VEGF DDS achieved a sustained 3-month delivery of antibodies to the anterior chamber, iris, ciliary body, and retina. Administration after corneal alkali burn suppressed CD45+ immune cell infiltration into the cornea, completely inhibited cornea NV for 3 months, accelerated corneal re-epithelialization and wound healing, and prevented RGC and optic nerve axon loss at 3 months. In contrast, anti-VEGF alone or IgG DDS treatment led to persistent corneal epithelial defect (combined: &lt;1%; anti-VEGF: 15%; IgG: 10%, of cornea area), increased infiltration of CD45+ immune cells into the cornea (combined: 28 \u00b1 20; anti-VEGF: 730 \u00b1 178; anti-IgG: 360 \u00b1 186, cells\/section), and significant loss of RGCs (combined: 2.7%; anti-VEGF: 63%; IgG: 45%) and optic nerve axons at 3 months. The aqueous humor protein analysis showed first-order release kinetics without adverse effects at the injection site. Conclusions: Concomitant inhibition of TNF-\u03b1 and VEGF prevents corneal neovascularization and ameliorates subsequent irreversible damage to the retina and optic nerve after severe ocular injury. A single subconjunctival administration of this therapy, using a biodegradable, slow-release thermosensitive DDS, achieved the sustained elution of therapeutic levels of antibodies to all ocular tissues for 3 months. This therapeutic approach has the potential to dramatically improve the outcomes of severe ocular injuries in patients and improve the therapeutic outcomes in patients with retinal vascular diseases.<\/jats:p>","DOI":"10.3390\/pharmaceutics15082059","type":"journal-article","created":{"date-parts":[[2023,7,31]],"date-time":"2023-07-31T09:22:38Z","timestamp":1690795358000},"page":"2059","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":30,"title":["Sustained Inhibition of VEGF and TNF-\u03b1 Achieves Multi-Ocular Protection and Prevents Formation of Blood Vessels after Severe Ocular Trauma"],"prefix":"10.3390","volume":"15","author":[{"given":"Chengxin","family":"Zhou","sequence":"first","affiliation":[{"name":"Department of Ophthalmology, Harvard Medical School, Boston, MA 02115, USA"},{"name":"Boston Keratoprosthesis Laboratory, Massachusetts Eye and Ear, Boston, MA 02114, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fengyang","family":"Lei","sequence":"additional","affiliation":[{"name":"Department of Ophthalmology, Harvard Medical School, Boston, MA 02115, USA"},{"name":"Boston Keratoprosthesis Laboratory, Massachusetts Eye and Ear, Boston, MA 02114, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6344-3085","authenticated-orcid":false,"given":"Jyoti","family":"Sharma","sequence":"additional","affiliation":[{"name":"Department of Ophthalmology, Harvard Medical School, Boston, MA 02115, USA"},{"name":"Boston Keratoprosthesis Laboratory, Massachusetts Eye and Ear, Boston, MA 02114, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pui-Chuen","family":"Hui","sequence":"additional","affiliation":[{"name":"Department of Ophthalmology, Harvard Medical School, Boston, MA 02115, USA"},{"name":"Boston Keratoprosthesis Laboratory, Massachusetts Eye and Ear, Boston, MA 02114, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Natalie","family":"Wolkow","sequence":"additional","affiliation":[{"name":"Department of Ophthalmology, Harvard Medical School, Boston, MA 02115, USA"},{"name":"David G. Cogan Laboratory of Eye Pathology and Ophthalmic Plastic Surgery Service, Massachusetts Eye and Ear, Boston, MA 02114, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Claes H.","family":"Dohlman","sequence":"additional","affiliation":[{"name":"Department of Ophthalmology, Harvard Medical School, Boston, MA 02115, USA"},{"name":"Boston Keratoprosthesis Laboratory, Massachusetts Eye and Ear, Boston, MA 02114, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8622-6478","authenticated-orcid":false,"given":"Demetrios G.","family":"Vavvas","sequence":"additional","affiliation":[{"name":"Department of Ophthalmology, Harvard Medical School, Boston, MA 02115, USA"},{"name":"Angiogenesis Laboratory, Massachusetts Eye and Ear, Boston, MA 02114, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7463-1599","authenticated-orcid":false,"given":"James","family":"Chodosh","sequence":"additional","affiliation":[{"name":"Department of Ophthalmology, Harvard Medical School, Boston, MA 02115, USA"},{"name":"Boston Keratoprosthesis Laboratory, Massachusetts Eye and Ear, Boston, MA 02114, USA"},{"name":"Disruptive Technology Laboratory, Massachusetts Eye and Ear, Boston, MA 02114, USA"},{"name":"Department of Ophthalmology and Visual Sciences, University of New Mexico School of Medicine, Albuquerque, NM 87108, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4544-4452","authenticated-orcid":false,"given":"Eleftherios I.","family":"Paschalis","sequence":"additional","affiliation":[{"name":"Department of Ophthalmology, Harvard Medical School, Boston, MA 02115, USA"},{"name":"Boston Keratoprosthesis Laboratory, Massachusetts Eye and Ear, Boston, MA 02114, USA"},{"name":"Disruptive Technology Laboratory, Massachusetts Eye and Ear, Boston, MA 02114, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2023,7,31]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"71","DOI":"10.1001\/archopht.126.1.71","article-title":"Promotion of graft survival by vascular endothelial growth factor a neutralization after high-risk corneal transplantation","volume":"126","author":"Bachmann","year":"2008","journal-title":"Arch. Ophthalmol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"114","DOI":"10.7205\/MILMED-D-16-00041","article-title":"Corneal and Corneoscleral Injury in Combat Ocular Trauma from Operations Iraqi Free-dom and Enduring Freedom","volume":"182","author":"Vlasov","year":"2017","journal-title":"Mil. Med."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1166","DOI":"10.1016\/S0161-6420(97)30167-5","article-title":"Post-traumatic proliferative vitreoretinopathy. The epidemiologic profile, onset, risk factors, and visual outcome","volume":"104","author":"Cardillo","year":"1997","journal-title":"Ophthalmology"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1322","DOI":"10.1097\/ICO.0b013e31821eead6","article-title":"Glaucoma in eyes with severe chemical burn, before and after keratoprosthesis","volume":"30","author":"Cade","year":"2011","journal-title":"Cornea"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"248","DOI":"10.1097\/ICO.0000000000001438","article-title":"Chemical Burns of the Eye: The Role of Retinal Injury and New Therapeutic Possibilities","volume":"37","author":"Dohlman","year":"2018","journal-title":"Cornea"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"349","DOI":"10.1097\/ICO.0000000000000067","article-title":"Glaucoma progression and role of glaucoma surgery in patients with Boston keratoprosthesis","volume":"33","author":"Crnej","year":"2014","journal-title":"Cornea"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"382","DOI":"10.1097\/ICO.0000000000000071","article-title":"Alkali burn to the eye: Protection using TNF-alpha inhibition","volume":"33","author":"Cade","year":"2014","journal-title":"Cornea"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"E11359","DOI":"10.1073\/pnas.1807123115","article-title":"Permanent neuroglial remodeling of the retina following infiltration of CSF1R inhibition-resistant peripheral monocytes","volume":"115","author":"Paschalis","year":"2018","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"336","DOI":"10.1007\/s00347-007-1512-2","article-title":"Inhibition of angiogenesis in the anterior chamber of the eye","volume":"104","author":"Bock","year":"2007","journal-title":"Ophthalmologe"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"804","DOI":"10.1136\/bjo.2006.107912","article-title":"Inhibition of experimental corneal neovascularisation by bevacizumab (Avastin)","volume":"91","author":"Manzano","year":"2007","journal-title":"Br. J. Ophthalmol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"862","DOI":"10.1111\/j.1755-3768.2009.01571.x","article-title":"The inhibitory effect of different concentrations of topical bevacizumab on corneal neovascularization","volume":"88","author":"Barequet","year":"2010","journal-title":"Acta Ophthalmol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"281","DOI":"10.1007\/s00417-007-0684-4","article-title":"Bevacizumab (Avastin) eye drops inhibit corneal neovascularization","volume":"246","author":"Bock","year":"2008","journal-title":"Graefes Arch. Clin. Exp. Ophthalmol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"e33","DOI":"10.1016\/j.ophtha.2008.02.013","article-title":"The effect of topical bevacizumab on corneal neovascularization","volume":"115","author":"Kim","year":"2008","journal-title":"Ophthalmology"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"381","DOI":"10.1001\/archophthalmol.2009.18","article-title":"Topical bevacizumab in the treatment of corneal neovascularization: Results of a prospective, open-label, noncomparative study","volume":"127","author":"Dastjerdi","year":"2009","journal-title":"Arch. Ophthalmol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"1580","DOI":"10.1016\/j.ajpath.2018.03.005","article-title":"The Role of Microglia and Peripheral Monocytes in Retinal Damage after Corneal Chemical Injury","volume":"188","author":"Paschalis","year":"2018","journal-title":"Am. J. Pathol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"569","DOI":"10.1038\/eye.2012.301","article-title":"Combined inhibition of tumor necrosis factor (TNF) and vascular endothelial growth factor (VEGF) for the treatment of macular edema of various etiologies: A short-term pilot study","volume":"27","author":"Arevalo","year":"2013","journal-title":"Eye"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"180","DOI":"10.1590\/S0004-27492013000300010","article-title":"Intravitreal bevacizumab combined with infliximab in the treatment of choroidal neovascularization secondary to age-related macular degeneration: Case report series","volume":"76","author":"Freitas","year":"2013","journal-title":"Arq. Bras. Oftalmol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"96","DOI":"10.1167\/iovs.16-20339","article-title":"Sustained Subconjunctival Delivery of Infliximab Protects the Cornea and Retina Following Alkali Burn to the Eye","volume":"58","author":"Zhou","year":"2017","journal-title":"Investig. Ophthalmol. Vis. Sci."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"4807","DOI":"10.1167\/iovs.08-3148","article-title":"Pharmacokinetics of bevacizumab after topical, subconjunctival, and intravitreal administration in rabbits","volume":"50","author":"Nomoto","year":"2009","journal-title":"Investig. Ophthalmol. Vis. Sci."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"67","DOI":"10.1016\/j.jtos.2012.01.005","article-title":"Corneal neovascularization and the utility of topical VEGF inhibition: Ranibizumab (Lucentis) vs bevacizumab (Avastin)","volume":"10","author":"Stevenson","year":"2012","journal-title":"Ocul. Surf."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"415","DOI":"10.1016\/j.survophthal.2012.01.007","article-title":"Corneal neovascularization: An anti-VEGF therapy review","volume":"57","author":"Chang","year":"2012","journal-title":"Surv. Ophthalmol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"42","DOI":"10.1016\/j.jconrel.2010.08.028","article-title":"Barrier analysis of periocular drug delivery to the posterior segment","volume":"148","author":"Ranta","year":"2010","journal-title":"J. Control Release"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"134","DOI":"10.1016\/j.preteyeres.2016.12.001","article-title":"Pharmacokinetic aspects of retinal drug delivery","volume":"57","author":"Rimpela","year":"2017","journal-title":"Prog. Retin. Eye Res."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"104501","DOI":"10.1016\/j.reactfunctpolym.2020.104501","article-title":"Recent advances in natural polymer-based drug delivery systems","volume":"148","author":"Tong","year":"2020","journal-title":"React. Funct. Polym."},{"key":"ref_25","unstructured":"Garner, J. (2021, August 28). Users Guide for Thermogelling PolyVivo PLGA-PEG-PLGA (AK12\/AK19\/AK24). Available online: https:\/\/akinainc.com\/pdf\/40307-PolyVivo-thermogel-usersguide.pdf."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"1327","DOI":"10.1016\/j.ajpath.2017.02.005","article-title":"Mechanisms of Retinal Damage after Ocular Alkali Burns","volume":"187","author":"Paschalis","year":"2017","journal-title":"Am. J. Pathol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"539","DOI":"10.4049\/jimmunol.1800982","article-title":"Microglia Regulate Neuroglia Remodeling in Various Ocular and Retinal Injuries","volume":"202","author":"Paschalis","year":"2019","journal-title":"J. Immunol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"2056","DOI":"10.1016\/j.ajpath.2020.07.006","article-title":"Glaucoma after ocular surgery or trauma: The role of infiltrating monocytes and their response to cytokine inhibitors","volume":"190","author":"Chen","year":"2020","journal-title":"Am. J. Pathol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"5","DOI":"10.1167\/tvst.9.8.5","article-title":"Microporous Drug Delivery System for Sustained Anti-VEGF Delivery to the Eye","volume":"9","author":"Zhou","year":"2020","journal-title":"Transl. Vis. Sci. Technol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"460","DOI":"10.1136\/bjophthalmol-2012-302435","article-title":"Intravitreal injections at the Massachusetts Eye and Ear Infirmary: Analysis of treatment indications and postinjection endophthalmitis rates","volume":"97","author":"Englander","year":"2013","journal-title":"Br. J. Ophthalmol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"11","DOI":"10.1167\/tvst.5.2.11","article-title":"A Drug Delivery System for Administration of Anti-TNF-\u03b1 Antibody","volume":"5","author":"Robert","year":"2016","journal-title":"Transl. Vis. Sci. Technol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"1589","DOI":"10.1097\/ICO.0000000000002106","article-title":"Glaucoma After Corneal Trauma or Surgery\u2014A Rapid, Inflammatory, IOP-Independent Pathway","volume":"38","author":"Dohlman","year":"2019","journal-title":"Cornea"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"375","DOI":"10.1016\/j.ijpharm.2015.05.071","article-title":"An injectable thermosensitive polymeric hydrogel for sustained release of Avastin\u00ae to treat posterior segment disease","volume":"490","author":"Xie","year":"2015","journal-title":"Int. J. Pharm."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"1375","DOI":"10.1007\/s00417-009-1099-1","article-title":"Short- and long-term safety profile and efficacy of topical bevacizumab (Avastin) eye drops against corneal neovascularization","volume":"247","author":"Koenig","year":"2009","journal-title":"Graefes Arch. Clin. Exp. Ophthalmol."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"6277","DOI":"10.1167\/iovs.09-4571","article-title":"The Different Effects of Early and Late Bevacizumab (Avastin) Injection on Inhibiting Corneal Neovascularization and Conjunctivalization in Rabbit Limbal Insufficiency","volume":"51","author":"Lin","year":"2010","journal-title":"Investig. Ophthalmol. Vis. Sci."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"1636","DOI":"10.1002\/art.20181","article-title":"Antiangiogenic effects of anti-tumor necrosis factor alpha therapy with infliximab in psoriatic arthritis","volume":"50","author":"Canete","year":"2004","journal-title":"Arthritis Rheum."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"R125","DOI":"10.1186\/ar2537","article-title":"Serum levels of biomarkers of bone and cartilage destruction and new bone formation in different cohorts of patients with axial spondyloarthritis with and without tumor necrosis factor-alpha blocker treatment","volume":"10","author":"Appel","year":"2008","journal-title":"Arthritis Res. Ther."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"439","DOI":"10.3109\/03009740903079321","article-title":"Effect of etanercept on serum levels of soluble cell adhesion molecules (sICAM-1, sVCAM-1, and sE-selectin) and vascular endothelial growth factor in patients with rheumatoid arthritis","volume":"38","author":"Klimiuk","year":"2009","journal-title":"Scand. J. Rheumatol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"113","DOI":"10.2217\/imt.12.151","article-title":"Effects of TNF-\u03b1 inhibitors upon the mechanisms of action of VEGF","volume":"5","author":"Murdaca","year":"2013","journal-title":"Immunotherapy"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"72","DOI":"10.1097\/ICL.0000000000000071","article-title":"Tumor Necrosis Factor-\u03b1 Inhibitors as a Treatment of Corneal Hemangiogenesis and Lymphangiogenesis","volume":"41","author":"Ferrari","year":"2014","journal-title":"Eye Contact Lens Sci. Clin. Pract."},{"key":"ref_41","first-page":"116","article-title":"TNF-\u03b1 mediates choroidal neovascularization by upregulating VEGF expression in RPE through ROS-dependent \u03b2-catenin activation","volume":"22","author":"Wang","year":"2016","journal-title":"Mol. Vis."},{"key":"ref_42","unstructured":"Paschalis, E.I., Robert, M.-C., and Dohlman, C.H. (2018). Treatment of Chemical Burn to the Eye: A Changing Picture, Springer."},{"key":"ref_43","doi-asserted-by":"crossref","unstructured":"Colby, K., and Dana, R. (2020). Foundations of Corneal Disease: Past, Present and Future, Springer International Publishing.","DOI":"10.1007\/978-3-030-25335-6"},{"key":"ref_44","doi-asserted-by":"crossref","unstructured":"Rezaeian Shiadeh, S.N., Hadizadeh, F., Khodaverdi, E., Gorji Valokola, M., Rakhshani, S., Kamali, H., and Nokhodchi, A. (2023). Injectable In-Situ Forming Depot Based on PLGA and PLGA-PEG-PLGA for Sustained-Release of Risperidone: In Vitro Evaluation and Pharmacokinetics in Rabbits. Pharmaceutics, 13.","DOI":"10.3390\/pharmaceutics15041229"}],"container-title":["Pharmaceutics"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1999-4923\/15\/8\/2059\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T20:23:09Z","timestamp":1760127789000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1999-4923\/15\/8\/2059"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,7,31]]},"references-count":44,"journal-issue":{"issue":"8","published-online":{"date-parts":[[2023,8]]}},"alternative-id":["pharmaceutics15082059"],"URL":"https:\/\/doi.org\/10.3390\/pharmaceutics15082059","relation":{},"ISSN":["1999-4923"],"issn-type":[{"value":"1999-4923","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,7,31]]}}}