{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,22]],"date-time":"2026-06-22T10:07:20Z","timestamp":1782122840321,"version":"3.54.5"},"reference-count":53,"publisher":"MDPI AG","issue":"8","license":[{"start":{"date-parts":[[2022,8,15]],"date-time":"2022-08-15T00:00:00Z","timestamp":1660521600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"National Research Foundation of South Africa Research","award":["64814"],"award-info":[{"award-number":["64814"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Pharmaceutics"],"abstract":"<jats:p>Delivering high-molecular-weight hydrophobic peptides, such as cyclosporine A, across the corneal epithelium remains a challenge that is complicated by other physio-anatomical ocular structures that limit the ocular bioavailability of such peptides. Transferosomes have previously been used to improve transdermal permeability, and have the potential for improving the ocular corneal permeability of applicable drugs. In this study, transferosomes for the potential ocular delivery of cyclosporine A were investigated. Linoleic acid was evaluated for its effect on the stability of the transferosomes and was substituted for a portion of the cholesterol in the vesicles. Additionally, Span\u00ae 80 and Tween\u00ae 80 were evaluated for their effect on transferosome flexibility and toxicity to ocular cells as edge activators. Attenuated Total Reflectance\u2013Fourier Transform Infrared spectroscopy (ATF-FTIR), differential scanning calorimetry (DSC), and dynamic light scattering (DLS) were used to evaluate the physicochemical parameters of the blank and the cyclosporine A-loaded transferosomes. Cyclosporine A release and corneal permeability were studied in vitro and in a New Zealand albino rabbit corneal model, respectively. The linoleic acid contributed to improved stability and the nano-size of the transferosomes. The Tween\u00ae-based formulation was preferred on the basis of a more favorable toxicity profile, as the difference in their corneal permeability was not significant. There was an initial burst release of cyclosporine A in the first 24 h that plateaued over one week. The Tween\u00ae-based formulation had a flux of 0.78 \u00b5g\/cm2\/h. The prepared transferosomes demonstrated biocompatibility in the ocular cell line, adequately encapsulated cyclosporine A, ensured the corneal permeability of the enclosed drug, and were stable over the period of investigation of 4 months at \u221220 \u00b0C.<\/jats:p>","DOI":"10.3390\/pharmaceutics14081695","type":"journal-article","created":{"date-parts":[[2022,8,15]],"date-time":"2022-08-15T03:33:20Z","timestamp":1660534400000},"page":"1695","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":35,"title":["Linoleic Acid-Based Transferosomes for Topical Ocular Delivery of Cyclosporine A"],"prefix":"10.3390","volume":"14","author":[{"given":"Onyinye","family":"Uwaezuoke","sequence":"first","affiliation":[{"name":"Wits Advanced Drug Delivery Platform Research Unit, Department of Pharmacy and Pharmacology, School of Therapeutic Sciences, Faculty of Health Sciences, University of the Witwatersrand, 7 York Road, Parktown, Johannesburg 2193, South Africa"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lisa C.","family":"Du Toit","sequence":"additional","affiliation":[{"name":"Wits Advanced Drug Delivery Platform Research Unit, Department of Pharmacy and Pharmacology, School of Therapeutic Sciences, Faculty of Health Sciences, University of the Witwatersrand, 7 York Road, Parktown, Johannesburg 2193, South Africa"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8640-4350","authenticated-orcid":false,"given":"Pradeep","family":"Kumar","sequence":"additional","affiliation":[{"name":"Wits Advanced Drug Delivery Platform Research Unit, Department of Pharmacy and Pharmacology, School of Therapeutic Sciences, Faculty of Health Sciences, University of the Witwatersrand, 7 York Road, Parktown, Johannesburg 2193, South Africa"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Naseer","family":"Ally","sequence":"additional","affiliation":[{"name":"Department of Neurosciences, Division of Ophthalmology, University of the Witwatersrand, 7 York Road, Parktown, Johannesburg 2193, South Africa"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3889-1529","authenticated-orcid":false,"given":"Yahya E.","family":"Choonara","sequence":"additional","affiliation":[{"name":"Wits Advanced Drug Delivery Platform Research Unit, Department of Pharmacy and Pharmacology, School of Therapeutic Sciences, Faculty of Health Sciences, University of the Witwatersrand, 7 York Road, Parktown, Johannesburg 2193, South Africa"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,8,15]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1903","DOI":"10.1007\/s13346-020-00879-1","article-title":"Fouling in ocular devices: Implications for drug delivery, bioactive surface immobilization, and biomaterial design","volume":"11","author":"Uwaezuoke","year":"2021","journal-title":"Drug Deliv. Transl. Res."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"3556","DOI":"10.1021\/mp500332m","article-title":"On the Barrier Properties of the Cornea: A Microscopy Study of the Penetration of Fluorescently Labeled Nanoparticles, Polymers, and Sodium Fluorescein","volume":"11","author":"Mun","year":"2014","journal-title":"Mol. Pharm."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"3","DOI":"10.1002\/jps.2600770103","article-title":"Mechanisms of corneal drug penetration. I: In vivo and in vitro kinetics","volume":"77","author":"Grass","year":"1988","journal-title":"J. Pharm. Sci."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"83","DOI":"10.1016\/j.ejps.2018.03.034","article-title":"Corneal and conjunctival drug permeability: Systematic comparison and pharmacokinetic impact in the eye","volume":"119","author":"Ramsay","year":"2018","journal-title":"Eur. J. Pharm. Sci."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"257","DOI":"10.1016\/j.ejpb.2012.02.014","article-title":"Novel micelle carriers for cyclosporin A topical ocular delivery: In vivo cornea penetration, ocular distribution and efficacy studies","volume":"81","author":"Bourges","year":"2012","journal-title":"Eur. J. Pharm. Biopharm."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1606","DOI":"10.1016\/j.carbpol.2011.09.065","article-title":"Thiolated pectin nanoparticles: Preparation, characterization and ex vivo corneal permeation study","volume":"87","author":"Sharma","year":"2012","journal-title":"Carbohydr. Polym."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"41","DOI":"10.1016\/S0378-5173(97)00083-5","article-title":"Evaluation of cationic polymer-coated nanocapsules as ocular drug carriers","volume":"153","author":"Calvo","year":"1997","journal-title":"Int. J. Pharm."},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Ibrahim, M.M., Maria, D.N., Wang, X., Simpson, R.N., Hollingsworth, T.J., and Jablonski, M.M. (2020). Enhanced Corneal Penetration of a Poorly Permeable Drug Using Bioadhesive Multiple Microemulsion Technology. Pharmaceutics, 12.","DOI":"10.3390\/pharmaceutics12080704"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"435","DOI":"10.1016\/0039-6257(93)90141-S","article-title":"Minimizing systemic absorption of topically administered ophthalmic drugs","volume":"37","author":"Urtti","year":"1993","journal-title":"Surv. Ophthalmol."},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Moiseev, R.V., Morrison, P.W.J., Steele, F., and Khutoryanskiy, V.V. (2019). Penetration Enhancers in Ocular Drug Delivery. Pharmaceutics, 11.","DOI":"10.3390\/pharmaceutics11070321"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"14","DOI":"10.1016\/j.ejpb.2017.03.006","article-title":"Cyclosporine A delivery to the eye: A comprehensive review of academic and industrial efforts","volume":"117","author":"Lallemand","year":"2017","journal-title":"Eur. J. Pharm. Biopharm."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"352","DOI":"10.3109\/09273948.2010.498657","article-title":"Clinical Review: Topical Ophthalmic Use of Cyclosporin A","volume":"18","author":"Utine","year":"2010","journal-title":"Ocul. Immunol. Inflamm."},{"key":"ref_13","unstructured":"Daull, P., Baudouin, C., Liang, H., Feraille, L., Barabino, S., and Garrigue, J.-S. (2021). Review of Preclinical Outcomes of a Topical Cationic Emulsion of Cyclosporine A for the Treatment of Ocular Surface Diseases. Ocul. Immunol. Inflamm., 1\u201311."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.jconrel.2020.01.057","article-title":"Advances and limitations of drug delivery systems formulated as eye drops","volume":"321","author":"Jumelle","year":"2020","journal-title":"J. Control. Release"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"15","DOI":"10.1016\/0039-6257(80)90072-7","article-title":"Corneal cytotoxicity of topically applied drugs, vehicles and preservatives","volume":"25","author":"Burstein","year":"1980","journal-title":"Surv. Ophthalmol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"226","DOI":"10.1016\/0005-2736(92)90154-E","article-title":"Lipid vesicles penetrate into intact skin owing to the transdermal osmotic gradients and hydration force","volume":"1104","author":"Cevc","year":"1992","journal-title":"Biochim. Biophys. Acta BBA-Biomembr."},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Opatha, S.A.T., Titapiwatanakun, V., and Chutoprapat, R. (2020). Transfersomes: A Promising Nanoencapsulation Technique for Transdermal Drug Delivery. Pharmaceutics, 12.","DOI":"10.3390\/pharmaceutics12090855"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"74","DOI":"10.1016\/j.jtos.2019.10.001","article-title":"Evaluation of use of essential fatty acids in topical ophthalmic preparations for dry eye","volume":"18","author":"Mudgil","year":"2020","journal-title":"Ocul. Surf."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"748","DOI":"10.1016\/j.ajo.2014.12.019","article-title":"Importance of tear film instability in dry eye disease in office workers using visual display terminals: The Osaka study","volume":"159","author":"Yokoi","year":"2015","journal-title":"Am. J. Ophthalmol."},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Hashmat, D., Shoaib, M.H., Ali, F.R., and Siddiqui, F. (2020). Lornoxicam controlled release transdermal gel patch: Design, characterization and optimization using co-solvents as penetration enhancers. PLoS ONE, 15.","DOI":"10.1371\/journal.pone.0228908"},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Sydykov, B., Oldenhof, H., Sieme, H., and Wolkers, W.F. (2018). Storage stability of liposomes stored at elevated subzero temperatures in DMSO\/sucrose mixtures. PLoS ONE, 13.","DOI":"10.1371\/journal.pone.0199867"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1013","DOI":"10.1081\/DDC-120025458","article-title":"Transfersomes\u2014A Novel Vesicular Carrier for Enhanced Transdermal Delivery: Development, Characterization, and Performance Evaluation","volume":"29","author":"Jain","year":"2003","journal-title":"Drug Dev. Ind. Pharm."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"888","DOI":"10.1080\/10717544.2018.1458923","article-title":"Improving the topical ocular pharmacokinetics of lyophilized cyclosporine A-loaded micelles: Formulation, in vitro and in vivo studies","volume":"25","author":"Yu","year":"2018","journal-title":"Drug Deliv."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"178","DOI":"10.3109\/03639045.2010.502534","article-title":"Standardized in vitro drug release test for colloidal drug carriers using modified USP dissolution apparatus I","volume":"37","author":"Lamprecht","year":"2011","journal-title":"Drug Dev. Ind. Pharm."},{"key":"ref_25","unstructured":"(2020, November 11). Development of a Convenient Ex Vivo Model for the Study of the Transcorneal Permeation of Drugs: Histological and Permeability Evaluation-Pescina-2015-Journal of Pharmaceutical Sciences-Wiley Online Library. Available online: https:\/\/onlinelibrary.wiley.com\/doi\/full\/10.1002\/jps.24231."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"e01088","DOI":"10.1016\/j.heliyon.2018.e01088","article-title":"Probing nanoliposomes using single particle analytical techniques: Effect of excipients, solvents, phase transition and zeta potential","volume":"4","author":"Danaei","year":"2018","journal-title":"Heliyon"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"24354","DOI":"10.1038\/s41598-021-03841-4","article-title":"Controlled rate slow freezing with lyoprotective agent to retain the integrity of lipid nanovesicles during lyophilization","volume":"11","author":"Yang","year":"2021","journal-title":"Sci. Rep."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"861","DOI":"10.1016\/j.jsps.2016.12.008","article-title":"Effect of cryoprotection on particle size stability and preservation of chitosan nanoparticles with and without hyaluronate or alginate coating","volume":"25","author":"Almalik","year":"2017","journal-title":"Saudi Pharm. J. SPJ"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"e683153","DOI":"10.1155\/2014\/683153","article-title":"Cyclosporine A Loaded PLGA Nanoparticles for Dry Eye Disease: In Vitro Characterization Studies","volume":"2014","author":"Wagh","year":"2014","journal-title":"J. Nanotechnol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"159","DOI":"10.1080\/08982100802310261","article-title":"Differential Scanning Calorimetry (DSC): A Tool to Study the Thermal Behavior of Lipid Bilayers and Liposomal Stability","volume":"18","author":"Demetzos","year":"2008","journal-title":"J. Liposome Res."},{"key":"ref_31","doi-asserted-by":"crossref","unstructured":"Londo\u00f1o, C.A., Rojas, J., Yarce, C.J., and Salamanca, C.H. (2019). Design of Prototype Formulations for In Vitro Dermal Delivery of the Natural Antioxidant Ferulic Acid Based on Ethosomal Colloidal Systems. Cosmetics, 6.","DOI":"10.3390\/cosmetics6010005"},{"key":"ref_32","unstructured":"(2022, June 16). The Merck Index: An Encyclopedia of Chemicals, Drugs, and Biologicals (Book, 2006) [WorldCat.org]. Available online: https:\/\/www.worldcat.org\/title\/merck-index-an-encyclopedia-of-chemicals-drugs-and-biologicals\/oclc\/70882070."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"10866","DOI":"10.1021\/acsomega.9b00825","article-title":"Liposomal Nanovesicles for Efficient Encapsulation of Staphylococcal Antibiotics","volume":"4","author":"Syed","year":"2019","journal-title":"ACS Omega"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"120640","DOI":"10.1016\/j.ijpharm.2021.120640","article-title":"Formulation development and upscaling of lipid nanocapsules as a drug delivery system for a novel cyclic GMP analogue intended for retinal drug delivery","volume":"602","author":"Urimi","year":"2021","journal-title":"Int. J. Pharm."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"17910","DOI":"10.1038\/s41598-020-74609-5","article-title":"Cell morphology maintenance in Bacillus subtilis through balanced peptidoglycan synthesis and hydrolysis","volume":"10","author":"Sassine","year":"2020","journal-title":"Sci. Rep."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"999","DOI":"10.1097\/00007890-199511150-00021","article-title":"Liposomal cyclosporine. Characterization of drug incorporation and interbilayer exchange","volume":"60","author":"Ouyang","year":"1995","journal-title":"Transplantation"},{"key":"ref_37","first-page":"139","article-title":"Oral cyclosporine A\u2014The current picture of its liposomal and other delivery systems","volume":"14","author":"Czogalla","year":"2008","journal-title":"Cell. Mol. Biol. Lett."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"607","DOI":"10.1007\/s11095-013-1184-3","article-title":"In vitro, in vivo, and in silico evaluation of the bioresponsive behavior of an intelligent intraocular implant","volume":"31","author":"Carmichael","year":"2014","journal-title":"Pharm. Res."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"34634","DOI":"10.1039\/C8RA07601D","article-title":"Nanocharacterization of liposomes for the encapsulation of water soluble compounds from Cordyceps sinensis CS1197 by a supercritical gas anti-solvent technique","volume":"8","author":"Shashidhar","year":"2018","journal-title":"RSC Adv."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"119","DOI":"10.1016\/j.ijpharm.2018.01.019","article-title":"Semifluorinated alkane based systems for enhanced corneal penetration of poorly soluble drugs","volume":"538","author":"Agarwal","year":"2018","journal-title":"Int. J. Pharm."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"438","DOI":"10.3390\/pharmaceutics7040438","article-title":"Transdermal Drug Delivery: Innovative Pharmaceutical Developments Based on Disruption of the Barrier Properties of the Stratum Corneum","volume":"7","author":"Alkilani","year":"2015","journal-title":"Pharmaceutics"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"1040","DOI":"10.1016\/j.jsps.2017.01.006","article-title":"Formulation and characterization of novel soft nanovesicles for enhanced transdermal delivery of eprosartan mesylate","volume":"25","author":"Ahad","year":"2017","journal-title":"Saudi Pharm. J."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"705886","DOI":"10.3389\/fbioe.2021.705886","article-title":"Liposomes: Structure, Biomedical Applications, and Stability Parameters With Emphasis on Cholesterol","volume":"9","author":"Nakhaei","year":"2021","journal-title":"Front. Bioeng. Biotechnol."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"192","DOI":"10.1016\/j.nano.2008.09.003","article-title":"Degradable Poly(\u03b2-amino ester) nanoparticles for cancer cytoplasmic drug delivery","volume":"5","author":"Shen","year":"2009","journal-title":"Nanomed. Nanotechnol. Biol. Med."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"7800","DOI":"10.1021\/acs.jctc.0c00609","article-title":"Three-Dimensional Packing Defects in Lipid Membrane as a Function of Membrane Order","volume":"16","author":"Tripathy","year":"2020","journal-title":"J. Chem. Theory Comput."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"2910","DOI":"10.1073\/pnas.1702065114","article-title":"Formation and size distribution of self-assembled vesicles","volume":"114","author":"Huang","year":"2017","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"49","DOI":"10.1016\/j.fbio.2015.12.002","article-title":"Vitamin A palmitate-bearing nanoliposomes: Preparation and characterization","volume":"13","author":"Pezeshky","year":"2016","journal-title":"Food Biosci."},{"key":"ref_48","doi-asserted-by":"crossref","unstructured":"Huang, Y., Tao, Q., Hou, D., Hu, S., Tian, S., Chen, Y., Gui, R., Yang, L., and Wang, Y. (2018, March 27). A Novel Ion-Exchange Carrier Based upon Liposome-Encapsulated Montmorillonite for Ophthalmic Delivery of Betaxolol Hydrochloride. Available online: https:\/\/www.dovepress.com\/a-novel-ion-exchange-carrier-based-upon-liposome-encapsulated--montmor-peer-reviewed-article-IJN.","DOI":"10.2147\/IJN.S122747"},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"100","DOI":"10.1016\/S0005-2736(01)00372-8","article-title":"Phase behavior and glass transition of 1,2-dioleoylphosphatidylethanolamine (DOPE) dehydrated in the presence of sucrose","volume":"1514","author":"Shalaev","year":"2001","journal-title":"Biochim. Biophys. Acta BBA-Biomembr."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"486","DOI":"10.1007\/s13346-019-00693-4","article-title":"Ultraflexible lipid vesicles allow topical absorption of cyclosporin A","volume":"10","author":"Carreras","year":"2020","journal-title":"Drug Deliv. Transl. Res."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"1003","DOI":"10.1021\/acsomega.9b02616","article-title":"Design and Characterization of Cyclosporine A-Loaded Nanofibers for Enhanced Drug Dissolution","volume":"5","author":"Dubey","year":"2020","journal-title":"ACS Omega"},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"411","DOI":"10.1089\/10807680260362696","article-title":"Blood concentrations of cyclosporin a during long-term treatment with cyclosporin a ophthalmic emulsions in patients with moderate to severe dry eye disease","volume":"18","author":"Small","year":"2002","journal-title":"J. Ocul. Pharmacol. Ther. Off. J. Assoc. Ocul. Pharmacol. Ther."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"756","DOI":"10.1021\/mp3005963","article-title":"Cyclodextrin-Mediated Enhancement of Riboflavin Solubility and Corneal Permeability","volume":"10","author":"Morrison","year":"2013","journal-title":"Mol. Pharm."}],"container-title":["Pharmaceutics"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1999-4923\/14\/8\/1695\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T00:08:43Z","timestamp":1760141323000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1999-4923\/14\/8\/1695"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,8,15]]},"references-count":53,"journal-issue":{"issue":"8","published-online":{"date-parts":[[2022,8]]}},"alternative-id":["pharmaceutics14081695"],"URL":"https:\/\/doi.org\/10.3390\/pharmaceutics14081695","relation":{},"ISSN":["1999-4923"],"issn-type":[{"value":"1999-4923","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,8,15]]}}}