{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,30]],"date-time":"2026-09-30T03:29:01Z","timestamp":1790738941129,"version":"4.1.0"},"reference-count":27,"publisher":"MDPI AG","issue":"10","license":[{"start":{"date-parts":[[2026,5,19]],"date-time":"2026-05-19T00:00:00Z","timestamp":1779148800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["JCM"],"abstract":"<jats:p>Purpose: To evaluate the clinical efficacy and safety of eye drops containing lyophilized amniotic membrane (AM) in the treatment of dry eye disease (DED), with a focus on tear film stabilization and epithelial\u2013immune balance. Methods: In this prospective cohort study, 40 patients (80 eyes) with DED were followed over six visits. The primary outcome was tear break-up time (TBUT). Secondary outcomes included corneal and conjunctival staining graded by the Oxford scale, meibomian gland parameters, corneal sensitivity (Cochet\u2013Bonnet esthesiometry), best-corrected visual acuity, intraocular pressure (IOP), and Schirmer I test. Continuous variables were analyzed using repeated-measures ANOVA with Greenhouse\u2013Geisser correction and Bonferroni post hoc testing; ordinal outcomes were analyzed using the Friedman test with Dunn\u2013Bonferroni correction. Results: TBUT increased significantly in both eyes (OD: +5.3 s; OS: +4.9 s; both p &lt; 0.001; \u03b7p2 \u2248 0.33). Corneal and conjunctival staining scores decreased (p &lt; 0.001), meibomian gland quality and expressibility improved (p &lt; 0.001), and corneal sensitivity increased (p &lt; 0.001), while visual acuity and IOP remained stable. Schirmer I values showed no significant change. The combined pattern of changes (TBUT \u2191, staining \u2193, meibum\/expressibility \u2191, sensitivity \u2191) indicates tear film stabilization and ocular surface improvement with a preserved safety profile. Conclusions: Lyophilized AM eye drops significantly prolong TBUT and improve clinical signs of DED, presumably by restoring the extracellular matrix (ECM) niche and the heavy-chain hyaluronan\/pentraxin 3 (HC-HA\/PTX3) complex, reducing proteolytic burden, and promoting a pro-resolving immune balance, with potential neurotrophic effects. These findings support the adjunctive use of AM-derived eye drops within contemporary TFOS DEWS II-based management algorithms for dry eye disease.<\/jats:p>","DOI":"10.3390\/jcm15103920","type":"journal-article","created":{"date-parts":[[2026,5,19]],"date-time":"2026-05-19T15:17:23Z","timestamp":1779203843000},"page":"3920","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Lyophilized Amniotic Membrane Eye Drops Stabilize the Tear Film in Dry Eye Disease: A Prospective Cohort Study"],"prefix":"10.3390","volume":"15","author":[{"given":"Jelena","family":"Kostic","sequence":"first","affiliation":[{"name":"Clinic for Eye Diseases, University Clinical Center of Serbia, 11000 Belgrade, Serbia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Svetlana","family":"Stanojlovic","sequence":"additional","affiliation":[{"name":"Clinic for Eye Diseases, University Clinical Center of Serbia, 11000 Belgrade, Serbia"},{"name":"Faculty of Medicine, University of Belgrade, 11000 Belgrade, Serbia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Natasa","family":"Maksimovic","sequence":"additional","affiliation":[{"name":"Institute of Epidemiology, Faculty of Medicine, University of Belgrade, 11000 Belgrade, Serbia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0001-0091-8490","authenticated-orcid":false,"given":"Vladimir","family":"Milutinovic","sequence":"additional","affiliation":[{"name":"Clinic for Eye Diseases, University Clinical Center of Serbia, 11000 Belgrade, Serbia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nada","family":"Avram","sequence":"additional","affiliation":[{"name":"Faculty of Medicine Fo\u010da, University of East Sarajevo, 73300 Fo\u010da, Bosnia and Herzegovina"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4321-3615","authenticated-orcid":false,"given":"Tanja","family":"Kalezic","sequence":"additional","affiliation":[{"name":"Clinic for Eye Diseases, University Clinical Center of Serbia, 11000 Belgrade, Serbia"},{"name":"Faculty of Medicine, University of Belgrade, 11000 Belgrade, Serbia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bojana","family":"Dacic Krnjaja","sequence":"additional","affiliation":[{"name":"Clinic for Eye Diseases, University Clinical Center of Serbia, 11000 Belgrade, Serbia"},{"name":"Faculty of Medicine, University of Belgrade, 11000 Belgrade, Serbia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9980-2588","authenticated-orcid":false,"given":"Borivoje","family":"Savic","sequence":"additional","affiliation":[{"name":"Clinic for Eye Diseases, University Clinical Center of Serbia, 11000 Belgrade, Serbia"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,5,19]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"276","DOI":"10.1016\/j.jtos.2017.05.008","article-title":"TFOS DEWS II definition and classification report","volume":"15","author":"Craig","year":"2017","journal-title":"Ocul. Surf."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"438","DOI":"10.1016\/j.jtos.2017.05.011","article-title":"TFOS DEWS II pathophysiology report","volume":"15","author":"Bron","year":"2017","journal-title":"Ocul. Surf."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"ORSFh1","DOI":"10.1167\/iovs.15-17637","article-title":"HC-HA\/PTX3 purified from amniotic membrane as novel regenerative matrix: Insight into relationship between inflammation and regeneration","volume":"57","author":"Tseng","year":"2016","journal-title":"Investig. Ophthalmol. Vis. Sci."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"42195","DOI":"10.1038\/srep42195","article-title":"Heavy Chain-Hyaluronan\/Pentraxin 3 from Amniotic Membrane Suppresses Inflammation and Scarring in Murine Lacrimal Gland and Conjunctiva of Chronic Graft-versus-Host Disease","volume":"7","author":"Ogawa","year":"2017","journal-title":"Sci. Rep."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"62","DOI":"10.1167\/iovs.61.5.62","article-title":"HC-HA\/PTX3 purified from human amniotic membrane reverts human corneal fibroblasts and myofibroblasts to keratocytes by activating BMP signaling","volume":"61","author":"Zhu","year":"2020","journal-title":"Investig. Ophthalmol. Vis. Sci."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1105","DOI":"10.2147\/OPTH.S165553","article-title":"Amniotic membrane extract and eye drops: A review of literature and clinical application","volume":"12","author":"Murri","year":"2018","journal-title":"Clin. Ophthalmol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"431","DOI":"10.1097\/ICO.0b013e31803dcda2","article-title":"Interleukin-6 and tumor necrosis factor-alpha levels in tears of patients with dry eye syndrome","volume":"26","author":"Yoon","year":"2007","journal-title":"Cornea"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1023","DOI":"10.1097\/ICO.0b013e3181a16578","article-title":"Analysis of inflammatory cytokines in the tears of dry eye patients","volume":"28","author":"Massingale","year":"2009","journal-title":"Cornea"},{"key":"ref_9","first-page":"862","article-title":"Tear cytokine and chemokine analysis and clinical correlations in evaporative-type dry eye disease","volume":"16","author":"Castellanos","year":"2010","journal-title":"Mol. Vis."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"5443","DOI":"10.1167\/iovs.11-9417","article-title":"Correlations between tear cytokines, chemokines, and soluble receptors and clinical severity of dry eye disease","volume":"53","author":"Na","year":"2012","journal-title":"Investig. Ophthalmol. Vis. Sci."},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Roda, M., Corazza, I., Bacchi Reggiani, M.L., Pellegrini, M., Taroni, L., Giannaccare, G., and Versura, P. (2020). Dry eye disease and tear cytokine levels\u2014A meta-analysis. Int. J. Mol. Sci., 21.","DOI":"10.3390\/ijms21093111"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"159","DOI":"10.1016\/j.ajpath.2015.09.012","article-title":"IL-10 Indirectly Regulates Corneal Lymphangiogenesis and Resolution of Inflammation via Macrophages","volume":"186","author":"Hos","year":"2016","journal-title":"Am. J. Pathol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"14","DOI":"10.1097\/ICL.0000000000001136","article-title":"Spotlight on Amniotic Membrane Extract Eye Drops: A Review of the Literature","volume":"51","author":"Kilian","year":"2025","journal-title":"Eye Contact Lens"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"855","DOI":"10.1111\/j.1442-9071.2009.02159.x","article-title":"Amniotic membrane extraction solution for ocular chemical burns","volume":"37","author":"Liang","year":"2009","journal-title":"Clin. Exp. Ophthalmol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"2973","DOI":"10.1007\/s10792-021-01856-4","article-title":"Amniotic membrane extract eye drops for ocular surface diseases: Use and clinical outcome in real-world practice","volume":"41","year":"2021","journal-title":"Int. Ophthalmol."},{"key":"ref_16","first-page":"2283","article-title":"Pro- and anti-inflammatory forms of interleukin-1 in the tear fluid and conjunctiva of patients with dry-eye disease","volume":"42","author":"Solomon","year":"2001","journal-title":"Investig. Ophthalmol. Vis. Sci."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"539","DOI":"10.1016\/j.jtos.2017.05.001","article-title":"TFOS DEWS II diagnostic methodology report","volume":"15","author":"Wolffsohn","year":"2017","journal-title":"Ocul. Surf."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"575","DOI":"10.1016\/j.jtos.2017.05.006","article-title":"TFOS DEWS II management and therapy report","volume":"15","author":"Jones","year":"2017","journal-title":"Ocul. Surf."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"366","DOI":"10.1016\/j.jtos.2017.03.006","article-title":"TFOS DEWS II tear film report","volume":"15","author":"Willcox","year":"2017","journal-title":"Ocul. Surf."},{"key":"ref_20","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_21","doi-asserted-by":"crossref","first-page":"19","DOI":"10.3109\/08830185.2012.748052","article-title":"Dry eye as a mucosal autoimmune disease","volume":"32","author":"Stern","year":"2013","journal-title":"Int. Rev. Immunol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"S4","DOI":"10.1016\/j.ophtha.2017.07.010","article-title":"The Pathophysiology of Dry Eye Disease: What We Know and Future Directions for Research","volume":"124","author":"Pflugfelder","year":"2017","journal-title":"Ophthalmology"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"379","DOI":"10.1016\/S0014-4835(03)00204-5","article-title":"Distribution of mucins at the ocular surface","volume":"78","author":"Gipson","year":"2004","journal-title":"Exp. Eye Res."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"88","DOI":"10.22203\/eCM.v015a07","article-title":"Properties of the amniotic membrane for potential use in tissue engineering","volume":"15","author":"Niknejad","year":"2008","journal-title":"Eur. Cells Mater."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"447","DOI":"10.1007\/s00441-012-1424-6","article-title":"Amniotic membrane: From structure and functions to clinical applications","volume":"349","author":"Mamede","year":"2012","journal-title":"Cell Tissue Res."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"51","DOI":"10.1016\/j.survophthal.2003.10.004","article-title":"The amniotic membrane in ophthalmology","volume":"49","author":"Dua","year":"2004","journal-title":"Surv. Ophthalmol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"640","DOI":"10.1097\/00003226-200310000-00008","article-title":"Grading of corneal and conjunctival staining in the context of other dry eye tests","volume":"22","author":"Bron","year":"2003","journal-title":"Cornea"}],"container-title":["Journal of Clinical Medicine"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2077-0383\/15\/10\/3920\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,5,19]],"date-time":"2026-05-19T15:35:23Z","timestamp":1779204923000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2077-0383\/15\/10\/3920"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,5,19]]},"references-count":27,"journal-issue":{"issue":"10","published-online":{"date-parts":[[2026,5]]}},"alternative-id":["jcm15103920"],"URL":"https:\/\/doi.org\/10.3390\/jcm15103920","relation":{},"ISSN":["2077-0383"],"issn-type":[{"value":"2077-0383","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,5,19]]}}}