{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,4]],"date-time":"2026-02-04T22:18:25Z","timestamp":1770243505268,"version":"3.49.0"},"reference-count":30,"publisher":"MDPI AG","issue":"2","license":[{"start":{"date-parts":[[2026,2,4]],"date-time":"2026-02-04T00:00:00Z","timestamp":1770163200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Scope Ophthalmics Ltd, the InnoHK initiative of the Innovation and Technology Commission of the Hong Kong Special Administrative Region Government, and Mitacs"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Pharmaceutics"],"abstract":"<jats:p>Background: Dry eye disease (DED) is characterized by tear film instability and a hyperosmolar ocular surface, which significantly impacts ocular health. Artificial tear solutions (ATSs) have been effective frontline treatments for DED, yet current commercially available products often provide only temporary relief, necessitating frequent daily administration. Significant efforts have been made to develop next-generation ATSs that can provide prolonged protective effects for DED. High-molecular-weight sodium hyaluronate (HA) is more commonly used in multi-dose preservative ATSs due to its longer chain lengths and rheological properties that can provide an enhanced retention time and clinical comfort and effects. The current methods to evaluate ATSs have largely focused on human biocompatibility and rheological testing and often overlook the dynamic nature of cellular phenotypes or the protective mechanisms at a cellular level. Therefore, this study developed novel in vitro mammalian cell assays involving human corneal epithelial cells (HCECs) to comprehensively assess ATSs with HA for biocompatibility and efficacy. Methods: We evaluated cellular viability across varying severities in two distinct DED models: desiccation and hyperosmotic stress. Simultaneously, time-lapse imaging coupled with computational image analyses quantified subtle, yet significant, cellular morphological changes under these stress condition. Results: Our assays revealed that ATSs provide significant, yet varying, protection against mild, medium, and harsh desiccation stress, as well as hyperosmotic conditions. This study also made a key insight that was the observation that DED conditions induce drastic HCEC morphological changes, including significant cellular monolayer breakage, which were effectively mitigated by the ATS products used in this work. Conclusions: The assays presented here provide a robust standard for ATS testing, ultimately guiding the selection of more effective next-generation therapies and aiding in a greater understanding of DED pathogenesis.<\/jats:p>","DOI":"10.3390\/pharmaceutics18020202","type":"journal-article","created":{"date-parts":[[2026,2,4]],"date-time":"2026-02-04T09:02:01Z","timestamp":1770195721000},"page":"202","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["An In Vitro Evaluation of the Effect and Protection of Artificial Tear Formulations on Human Corneal Epithelial Cells in Normal and Dry Eye Disease States"],"prefix":"10.3390","volume":"18","author":[{"given":"Ka-Ying","family":"Wong","sequence":"first","affiliation":[{"name":"Department of Chemistry, Waterloo Institute for Nanotechnology, University of Waterloo, Waterloo, ON N2L 3G1, Canada"},{"name":"Centre for Eye and Vision Research (CEVR), 17W Hong Kong Science Park, Hong Kong"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Brandon","family":"Ho","sequence":"additional","affiliation":[{"name":"Centre for Eye and Vision Research (CEVR), 17W Hong Kong Science Park, Hong Kong"},{"name":"Centre for Ocular Research & Education (CORE), School of Optometry & Vision Science, University of Waterloo, Waterloo, ON N2L 3G1, Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Asfia","family":"Soomro","sequence":"additional","affiliation":[{"name":"Centre for Ocular Research & Education (CORE), School of Optometry & Vision Science, University of Waterloo, Waterloo, ON N2L 3G1, Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7409-7349","authenticated-orcid":false,"given":"Lyndon","family":"Jones","sequence":"additional","affiliation":[{"name":"Centre for Eye and Vision Research (CEVR), 17W Hong Kong Science Park, Hong Kong"},{"name":"Centre for Ocular Research & Education (CORE), School of Optometry & Vision Science, University of Waterloo, Waterloo, ON N2L 3G1, Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5918-9336","authenticated-orcid":false,"given":"Juewen","family":"Liu","sequence":"additional","affiliation":[{"name":"Department of Chemistry, Waterloo Institute for Nanotechnology, University of Waterloo, Waterloo, ON N2L 3G1, Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Chau-Minh","family":"Phan","sequence":"additional","affiliation":[{"name":"Centre for Eye and Vision Research (CEVR), 17W Hong Kong Science Park, Hong Kong"},{"name":"Centre for Ocular Research & Education (CORE), School of Optometry & Vision Science, University of Waterloo, Waterloo, ON N2L 3G1, Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2026,2,4]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"334","DOI":"10.1016\/j.jtos.2017.05.003","article-title":"TFOS DEWS II Epidemiology Report","volume":"15","author":"Stapleton","year":"2017","journal-title":"Ocul. Surf."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"246","DOI":"10.1016\/j.jtos.2013.07.003","article-title":"Role of Hyperosmolarity in the Pathogenesis and Management of Dry Eye Disease: Proceedings of the OCEAN Group Meeting","volume":"11","author":"Baudouin","year":"2013","journal-title":"Ocul. Surf."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"248","DOI":"10.1111\/cei.12759","article-title":"Desiccating stress-induced disruption of ocular surface immune tolerance drives dry eye disease","volume":"184","author":"Keitelman","year":"2016","journal-title":"Clin. Exp. Immunol."},{"key":"ref_4","first-page":"163","article-title":"Persistent Corneal Epithelial Defects: A Review Article","volume":"8","author":"Vaidyanathan","year":"2019","journal-title":"Med. Hypothesis Discov. Innov. Ophthalmol."},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Agarwal, P., Craig, J.P., and Rupenthal, I.D. (2021). Formulation Considerations for the Management of Dry Eye Disease. Pharmaceutics, 13.","DOI":"10.3390\/pharmaceutics13020207"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"2283","DOI":"10.2147\/OPTH.S465143","article-title":"Tolerability of Current Treatments for Dry Eye Disease: A Review of Approved and Investigational Therapies","volume":"18","author":"Gupta","year":"2024","journal-title":"Clin. Ophthalmol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"1611","DOI":"10.3906\/sag-2002-143","article-title":"The negative effects of dry eye disease on quality of life and visual function","volume":"50","author":"Guo","year":"2020","journal-title":"Turk. J. Med. Sci."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"289","DOI":"10.1016\/j.ajo.2025.05.039","article-title":"TFOS DEWS III: Management and Therapy","volume":"279","author":"Jones","year":"2025","journal-title":"Am. J. Ophthalmol."},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Labetoulle, M., Benitez-del-Castillo, J.M., Barabino, S., Herrero Vanrell, R., Daull, P., Garrigue, J.-S., and Rolando, M. (2022). Artificial Tears: Biological Role of Their Ingredients in the Management of Dry Eye Disease. Int. J. Mol. Sci., 23.","DOI":"10.3390\/ijms23052434"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"9","DOI":"10.2147\/OPTO.S350185","article-title":"Artificial Tears: A Systematic Review","volume":"15","author":"Semp","year":"2023","journal-title":"Clin. Optom."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"114867","DOI":"10.1016\/j.addr.2023.114867","article-title":"Non-aqueous formulations in topical ocular drug delivery\u2014A paradigm shift?","volume":"198","author":"Agarwal","year":"2023","journal-title":"Adv. Drug Deliv. Rev."},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Yang, Y.-J., Lee, W.-Y., Kim, Y., and Hong, Y. (2021). A Meta-Analysis of the Efficacy of Hyaluronic Acid Eye Drops for the Treatment of Dry Eye Syndrome. Int. J. Environ. Res. Public Health, 18.","DOI":"10.3390\/ijerph18052383"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"e000697","DOI":"10.1136\/bmjophth-2020-000697","article-title":"Dry eye: Why artificial tears are not always the answer","volume":"6","author":"Kim","year":"2021","journal-title":"BMJ Open Ophthalmol."},{"key":"ref_14","first-page":"CD009729","article-title":"Over the counter (OTC) artificial tear drops for dry eye syndrome","volume":"2016","author":"Pucker","year":"2016","journal-title":"Cochrane Database Syst. Rev."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"49","DOI":"10.1016\/j.actbio.2018.11.020","article-title":"Bioinspired hydrogels for drug-eluting contact lenses","volume":"84","author":"Concheiro","year":"2019","journal-title":"Acta Biomater."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"3032","DOI":"10.1167\/iovs.06-1507","article-title":"Effect of Blinking on Tear Dynamics","volume":"48","author":"Palakuru","year":"2007","journal-title":"Investig. Ophthalmol. Vis. Sci."},{"key":"ref_17","first-page":"100577","article-title":"Effectiveness evaluation of hyaluronic acid-based commercial eye drops to treat ophthalmic dry eye disease","volume":"8","author":"Alonso","year":"2024","journal-title":"Carbohydr. Polym. Technol. Appl."},{"key":"ref_18","first-page":"102","article-title":"Hyaluronic acid and its biomedical applications: A review","volume":"1","author":"Dovedytis","year":"2020","journal-title":"Eng. Regen."},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Gupta, R.C., Lall, R., Srivastava, A., and Sinha, A. (2019). Hyaluronic Acid: Molecular Mechanisms and Therapeutic Trajectory. Front. Vet. Sci., 6.","DOI":"10.3389\/fvets.2019.00192"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"844","DOI":"10.1111\/aos.15159","article-title":"Hyaluronic acid in the treatment of dry eye disease","volume":"100","author":"Hynnekleiv","year":"2022","journal-title":"Acta Ophthalmol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"821","DOI":"10.1136\/bjo.2003.027573","article-title":"Sodium hyaluronate (hyaluronic acid) promotes migration of human corneal epithelial cells in vitro","volume":"88","author":"Gomes","year":"2004","journal-title":"Br. J. Ophthalmol."},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Kojima, T., Nagata, T., Kudo, H., M\u00fcller-Lierheim, W.G.K., Van Setten, G.-B., Dogru, M., and Tsubota, K. (2020). The Effects of High Molecular Weight Hyaluronic Acid Eye Drop Application in Environmental Dry Eye Stress Model Mice. Int. J. Mol. Sci., 21.","DOI":"10.3390\/ijms21103516"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"2","DOI":"10.1167\/tvst.8.6.2","article-title":"Physicochemical Properties of Hyaluronic Acid\u2013Based Lubricant Eye Drops","volume":"8","author":"Aragona","year":"2019","journal-title":"Transl. Vis. Sci. Technol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"207","DOI":"10.1007\/s11626-020-00533-7","article-title":"In vitro reconstructed 3D corneal tissue models for ocular toxicology and ophthalmic drug development","volume":"57","author":"Kaluzhny","year":"2021","journal-title":"Vitr. Cell. Dev. Biol.-Anim."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"32","DOI":"10.1016\/j.tox.2014.11.003","article-title":"An overview of current techniques for ocular toxicity testing","volume":"327","author":"Wilson","year":"2015","journal-title":"Toxicology"},{"key":"ref_26","first-page":"60812","article-title":"Effect of Artificial Tear Formulations on the Metabolic Activity of Human Corneal Epithelial Cells after Exposure to Desiccation","volume":"159","author":"Rangarajan","year":"2020","journal-title":"J. Vis. Exp."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"DES192","DOI":"10.1167\/iovs.17-23651","article-title":"Inflammatory Response in Dry Eye","volume":"59","author":"Yamaguchi","year":"2018","journal-title":"Investig. Ophthalmol. Vis. Sci."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"108723","DOI":"10.1016\/j.exer.2021.108723","article-title":"Combined hyperosmolarity and inflammatory conditions in stressed human corneal epithelial cells and macrophages to evaluate osmoprotective agents as potential DED treatments","volume":"211","year":"2021","journal-title":"Exp. Eye Res."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"66","DOI":"10.1208\/s12249-023-02516-9","article-title":"Ocular Drug Delivery: A Comprehensive Review","volume":"24","author":"Ahmed","year":"2023","journal-title":"AAPS PharmSciTech"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"90","DOI":"10.1001\/archophthalmol.2011.364","article-title":"Dry Eye Disease: An Immune-Mediated Ocular Surface Disorder","volume":"130","author":"Stevenson","year":"2012","journal-title":"Arch. Ophthalmol."}],"container-title":["Pharmaceutics"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1999-4923\/18\/2\/202\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,2,4]],"date-time":"2026-02-04T09:05:13Z","timestamp":1770195913000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1999-4923\/18\/2\/202"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,2,4]]},"references-count":30,"journal-issue":{"issue":"2","published-online":{"date-parts":[[2026,2]]}},"alternative-id":["pharmaceutics18020202"],"URL":"https:\/\/doi.org\/10.3390\/pharmaceutics18020202","relation":{},"ISSN":["1999-4923"],"issn-type":[{"value":"1999-4923","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,2,4]]}}}