{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T18:58:14Z","timestamp":1785437894531,"version":"3.56.0"},"reference-count":44,"publisher":"MDPI AG","issue":"6","license":[{"start":{"date-parts":[[2026,5,25]],"date-time":"2026-05-25T00:00:00Z","timestamp":1779667200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100010679","name":"Pasteur Institute of Iran","doi-asserted-by":"publisher","award":["BP-66001899"],"award-info":[{"award-number":["BP-66001899"]}],"id":[{"id":"10.13039\/501100010679","id-type":"DOI","asserted-by":"publisher"}]},{"award":["BP-66001899"],"award-info":[{"award-number":["BP-66001899"]}],"id":[{"id":"https:\/\/ror.org\/00wqczk30","id-type":"ROR","asserted-by":"publisher"}]},{"name":"Student Research Committee, Pasteur Institute of Iran","award":["2475"],"award-info":[{"award-number":["2475"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biomolecules"],"abstract":"<jats:p>Current treatments for choroidal neovascularization (CNV) and its associated subretinal fibrosis (SRF), major causes of vision loss, are limited by the need for frequent intravitreal injections and the emergence of drug resistance. This study evaluated the safety and efficacy of the intravitreal administration of engineered humanized anti-vascular endothelial growth factor Nanobodies, including a wild-type Nanobody (WHNb) and two mutated variants (MHNb136 and MHNb256), in a rat model of laser-induced CNV and associated SRF. Safety was assessed through in vivo electrophysiological and histopathological analyses following intravitreal injection of Nanobodies at doses of 12.5, 25, 50, and 100 \u00b5g. Efficacy was evaluated in rat models of laser-induced CNV and SRF using double immunohistochemistry for isolectin B4 and anti-collagen type I on sclerochoroidal flat mounts. Mean CNV and SRF areas in Nanobody-treated groups were compared with those in bevacizumab-treated and sham control groups. None of the Nanobodies showed retinal toxicity in safety assessments. Compared with bevacizumab, MHNb136 and MHNb256 reduced the CNV area by 1.72-fold and 1.8-fold, respectively (both p &lt; 0.0001), whereas WHNb showed an effect nearly identical to that of bevacizumab. In addition, 12.5 \u00b5g MHNb136 and 100 \u00b5g MHNb256 reduced the SRF area by 1.3-fold (p = 0.047) and 1.6-fold (p = 0.0007), respectively, relative to bevacizumab. For CNV reduction, 12.5 \u00b5g MHNb136 was comparable to 25 \u00b5g MHNb256; both outperformed bevacizumab. For SRF reduction, 12.5 \u00b5g MHNb136 was more effective than bevacizumab and comparable to 100 \u00b5g MHNb256. These findings suggest that 12.5 \u00b5g MHNb136 may represent a cost-effective bioengineered Nanobody candidate for future clinical studies.<\/jats:p>","DOI":"10.3390\/biom16060772","type":"journal-article","created":{"date-parts":[[2026,5,25]],"date-time":"2026-05-25T15:05:08Z","timestamp":1779721508000},"page":"772","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Ocular Safety of Intravitreal Engineered Humanized Anti-VEGF Nanobody and Its Efficacy in the Attenuation of Choroidal Neovascularization and Associated Subretinal Fibrosis"],"prefix":"10.3390","volume":"16","author":[{"given":"Mir Salar","family":"Kazemi","sequence":"first","affiliation":[{"name":"Venom and Biotherapeutics Molecules Laboratory, Biotechnology Research Center, Pasteur Institute of Iran, Tehran 13169-43551, Iran"},{"name":"Student Research Committee, Pasteur Institute of Iran, Tehran 13169-43551, Iran"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mozhgan","family":"Rezaei Kanavi","sequence":"additional","affiliation":[{"name":"Ocular Tissue Engineering Research Center, Research Institute for Ophthalmology and Vision Science, Shahid Beheshti University of Medical Sciences, Tehran 16666-73111, Iran"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5832-1822","authenticated-orcid":false,"given":"Fatemeh","family":"Kazemi-Lomedasht","sequence":"additional","affiliation":[{"name":"Venom and Biotherapeutics Molecules Laboratory, Biotechnology Research Center, Pasteur Institute of Iran, Tehran 13169-43551, Iran"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Reza","family":"Ahangari Cohan","sequence":"additional","affiliation":[{"name":"Nanobiotechnology Department, New Technologies Research Group, Pasteur Institute of Iran, Tehran 13169-43551, Iran"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Golnoosh","family":"Mahjoobi","sequence":"additional","affiliation":[{"name":"Venom and Biotherapeutics Molecules Laboratory, Biotechnology Research Center, Pasteur Institute of Iran, Tehran 13169-43551, Iran"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sare","family":"Safi","sequence":"additional","affiliation":[{"name":"Ophthalmic Epidemiology Research Center, Research Institute for Ophthalmology and Vision Science, Shahid Beheshti University of Medical Sciences, Tehran 16666-73111, Iran"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sadra","family":"Ashrafi","sequence":"additional","affiliation":[{"name":"Ocular Tissue Engineering Research Center, Research Institute for Ophthalmology and Vision Science, Shahid Beheshti University of Medical Sciences, Tehran 16666-73111, Iran"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hamid","family":"Ahmadieh","sequence":"additional","affiliation":[{"name":"Ophthalmic Research Center, Research Institute for Ophthalmology and Vision Science, Shahid Beheshti University of Medical Sciences, Tehran 16666-73111, Iran"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alireza","family":"Shoari","sequence":"additional","affiliation":[{"name":"Department of Cancer Biology, Mayo Clinic, Jacksonville, FL 32224, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mahdi","family":"Behdani","sequence":"additional","affiliation":[{"name":"Venom and Biotherapeutics Molecules Laboratory, Biotechnology Research Center, Pasteur Institute of Iran, Tehran 13169-43551, Iran"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,5,25]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"e1175","DOI":"10.1016\/S2214-109X(25)00143-3","article-title":"Global Burden of Vision Impairment Due to Age-Related Macular Degeneration, 1990\u20132021, with Forecasts to 2050: A Systematic Analysis for the Global Burden of Disease Study 2021","volume":"13","author":"Jeong","year":"2025","journal-title":"Lancet Glob. 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