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Molecular Advances in Dry Eye Syndrome

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A special issue of International Journal of Molecular Sciences (ISSN 1422-0067). This special issue belongs to the section "Molecular Pathology, Diagnostics, and Therapeutics".

Deadline for manuscript submissions: closed (20 June 2025) | Viewed by 15328

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Prof. Dr. Maria Jesus Giráldez-Fernández

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1. Applied Physics Department (Optometry Area), Facultade de Óptica e Optometría, Universidade de Santiago de Compostela, 15705 Santiago de Compostela, Spain
2. Optometry Group, Instituto de Investigación Sanitaria Santiago de Compostela (IDIS), 15706 Santiago de Compostela, Spain
Interests: optometry; dry eye disease; ocular surface; contact lens; tear substitutes; myopia; ophthalmic instrumentation
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Dr. Anxo Fernández Ferreiro

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Clinical Pharmacology Group, Health Research Institute of Santiago de Compostela (IDIS), 15706 Santiago de Compostela, Spain
Interests: ocular drug delivery; ocular pharmacokinetic; drug development; pharmacogenetics; clinical and translational research
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Dear Colleagues,

Dry eye disease (DED) is a common ocular surface disease affecting a significant percentage of the population. Caused by tear film homeostasis disruption, DED is characterized by dryness, inflammation, discomfort, and visual disturbances. This condition greatly affects the quality of life of patients, and can potentially lead to permanent vision loss.

The purpose of this Special Issue is to report recent molecular advances in DED to assist in developing a better knowledge of DED-related pathophysiological mechanisms, diagnostic methods, and therapeutic approaches.

Potential topics include, but are not limited to, both immune and adaptive immune responses, ocular surface signs and symptoms, diagnostic methods, new pharmacological treatments, and specialized drug delivery systems.

Dr. Maria Jesus Giráldez-Fernández
Dr. Anxo Fernández Ferreiro
Guest Editors

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Submitted manuscripts should not have been published previously, nor be under consideration for publication elsewhere (except conference proceedings papers). All manuscripts are thoroughly refereed through a single-blind peer-review process. A guide for authors and other relevant information for submission of manuscripts is available on the Instructions for Authors page. International Journal of Molecular Sciences is an international peer-reviewed open access semimonthly journal published by MDPI.

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Keywords

dry eye disease
tear film osmolarity
tear film
artificial tear substitutes
biological tear substitutes
tear fluid biomarkers
tear film lipid layer
meibomian gland dysfunction
evaporative dry eye
aqueous deficiency dry eye
ocular surface inflammation
cytokines
histological analysis
pharmacological treatment

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Related Special Issue

Molecular Advances in Dry Eye Syndrome, Second Edition in International Journal of Molecular Sciences

Published Papers (11 papers)

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Research

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25 pages, 570 KB

Open AccessArticle

Long-Term Clinical and Molecular Changes in Dry Eye Disease and Chronic Ocular Pain

by Cristina Valencia-Sandonís, Andrés Ángel Calderón-García, Marta Blanco-Vázquez, Laura Valencia-Nieto, Andrea Novo-Diez, Amanda Vázquez, Margarita Calonge, María J. González-García and Amalia Enríquez-de-Salamanca

Int. J. Mol. Sci. 2025, 26(18), 8918; https://doi.org/10.3390/ijms26188918 - 12 Sep 2025

Viewed by 421

Abstract

Dry eye disease (DED) is a prevalent condition characterized by ocular surface inflammation and pain. This study evaluated the long-term progression of DED by analyzing clinical and molecular status, considering the impact of chronic ocular pain. Patients with DED were evaluated at two visits (V1 and V2) separated by at least two years. Evaluations included validated symptom questionnaires alongside slit-lamp examination, corneal sensitivity testing, and sub-basal nerve plexus analysis. Basal tear samples were collected for multiplex quantification of 20 cytokines and substance P (SP), and conjunctival cells were obtained to analyze 25 genes and 12 microRNAs (miRNA). Based on the presence or absence of chronic ocular pain, patients were then divided into two groups. Patients improved in DED-related symptoms, with no changes observed in ocular surface signs. Corneal dendritic cell density decreased, along with epidermal growth factor (EGF), fractalkine, and monocyte chemoattractant protein (MCP-1) tear levels, whereas interleukin (IL)-10 and SP tear levels increased. Neurotrophic tyrosine kinase, receptor, type (NTRK)1 gene expression was significantly downregulated, especially in patients without chronic ocular pain. miR-665 expression decreased significantly in DED patients. Monitoring corneal dendritic cells, tear cytokines, and gene/miRNA expression offers promising tools for tracking DED progression. Distinguishing the presence of chronic ocular pain as a separate symptom is crucial to optimizing therapeutic strategies and DED progression. Full article

(This article belongs to the Special Issue Molecular Advances in Dry Eye Syndrome)

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24 pages, 4279 KB

Open AccessArticle

Effects of Exposure of PHMG-p, a Humidifier Disinfectant Component, on Eye Dryness: A Study on a Rat Model Based on ¹H-NMR Metabolomics

by Jung Dae Lee, Hyang Yeon Kim, Soo Bean Oh, Hyeyoon Goo, Kyong Jin Cho, Gi-Wook Hwang, Suhkmann Kim and Kyu-Bong Kim

Int. J. Mol. Sci. 2025, 26(17), 8660; https://doi.org/10.3390/ijms26178660 - 5 Sep 2025

Viewed by 975

Abstract

Polyhexamethylene guanidine phosphate (PHMG-p), a widely used disinfectant component in household humidifiers, has been implicated in various health issues, including pulmonary toxicity. Many people use humidifiers to improve dry eye disease (DED). The current study was performed to elucidate the effect of PHMG-p on eye dryness in a rat model using metabolomics. Male Sprague Dawley rats were exposed to PHMG-p (0.1% and 0.3%) following a previously established DED induction model using scopolamine hydrobromide and desiccation stress. Ocular surface damage was assessed using corneal fluorescein staining, tear volume measurement, and tear break-up time (TBUT). Plasma and urine samples were analyzed using ¹H-NMR-based metabolomics to identify metabolic alterations associated with PHMG-P-p exposure and DED pathogenesis. PHMG-p exposure exacerbated DED symptoms, as evidenced by a significant reduction in tear volume, shorter TBUT, and increased corneal damage compared to the control group. Metabolomic profiling identified distinct metabolic changes in PHMG-p-exposed groups, including alterations in glutamate, glycine, citrate, and succinate metabolism. These metabolic changes correlated with increased levels of inflammatory cytokines such as IL-1β, IL-6, and TNF-α in the corneal and lacrimal gland tissues. Our findings suggest that PHMG-p exposure contributes to DED pathophysiology by inducing metabolic disturbances and inflammatory responses in the ocular surface. This study highlights the need for further investigation into the potential risks of PHMG-p exposure on ocular health and provides novel insights into the metabolic underpinnings of DED. Full article

(This article belongs to the Special Issue Molecular Advances in Dry Eye Syndrome)

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14 pages, 1400 KB

Open AccessArticle

Potential Roles of Extracellular Vesicles in Murine Tear Fluids in the Physiology of Corneal Epithelial Cells In Vitro

by Saya Oya, Kazunari Higa, Tomohiro Yasutake, Risa Yamazaki-Hokama and Masatoshi Hirayama

Int. J. Mol. Sci. 2025, 26(15), 7559; https://doi.org/10.3390/ijms26157559 - 5 Aug 2025

Viewed by 411

Abstract

Biological extracellular vesicles in tear fluids, such as exosomes, are thought to have physiological functions in the management of healthy ocular surface epithelium, including corneal epithelium. However, the physiological roles of tear extracellular vesicles in the ocular surface remain unclear. In this study, we investigated the physiological function of tear extracellular vesicles in mouse tear fluids in the ocular surface epithelium in vitro. Morphological analysis of the isolated extracellular vesicles from mouse tear fluids was performed using nanoparticle tracking analysis and transmission electron microscopy. The identified particles were characterised by immunoblotting for exosomal markers. After confirming the uptake of tear exosomes in cultured corneal epithelial cells, gene expression changes in mouse cultured corneal epithelial cells after tear exosome treatment were analysed. Immunostaining analysis was performed to confirm cell proliferation in the cultured corneal epithelial cells with tear exosome treatment. Tear fluids from mice contain nanoparticles with exosome-like morphologies, which express the representative exosomal markers CD9 and TSG101. The extracellular vesicles can be taken up by cultivated murine corneal epithelial cells in vitro and induce expression changes in genes related to the cell cycle, cell membranes, microtubules, and signal peptides. Treatment with the tear extracellular vesicles promoted cell proliferation of cultured murine corneal epithelial cells. Our study provides evidence that murine tear fluids contain extracellular vehicles like exosomes and they may contribute to the maintenance of the physiological homeostatic environment of the ocular surface. Full article

(This article belongs to the Special Issue Molecular Advances in Dry Eye Syndrome)

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16 pages, 3484 KB

Open AccessArticle

Exosomes from Limosilactobacillus fermentum Ameliorate Benzalkonium Chloride-Induced Inflammation in Conjunctival Cells

by Kippeum Lee, Hyeonjun Gwon, Joo Yun Kim, Jae Jung Shim and Jae Hwan Lee

Int. J. Mol. Sci. 2024, 25(22), 12282; https://doi.org/10.3390/ijms252212282 - 15 Nov 2024

Cited by 2 | Viewed by 1955

Abstract

Dry eye is characterized by persistent instability and decreased tear production, which are accompanied by epithelial lesions and inflammation on the surface of the eye. In our previous paper, we reported that supplementation with Limosilactobacillus fermentum HY7302 (HY7302) could inhibit corneal damage in a benzalkonium chloride (BAC)-induced mouse model of dry eye, through its effects in gut microbiome regulation. The aim of this study was to determine what functional extracellular substances can alter the inflammatory response of conjunctival cells. We isolated exosomes from HY7302 probiotic culture supernatant, analyzed their morphological characteristics, and found that their average size was 143.8 ± 1.1 nm, which was smaller than the exosomes from the L. fermentum KCTC 3112 strain. In addition, HY7302-derived exosomes significantly reduced the levels of genes encoding pro-inflammatory cytokines, including interleukin (IL)-20, IL-8, IL-6, and IL-1B, in BAC-treated human conjunctival cells. Moreover, HY7302-derived exosomes significantly increased the levels of genes encoding tight junction proteins, including TJP1, TJP2, and occludin-1, in Caco-2 cells. Lastly, the HY7302 exosomes reduced mRNA expression levels of IL1B, IL20, IL6, IL8, and NFAT5 in a transwell coculture system. Our findings indicate that HY7302 exosomes have potential for use in the treatment of ocular inflammation-related dry eye disease, through gut–eye axis communication via exosomes. Full article

(This article belongs to the Special Issue Molecular Advances in Dry Eye Syndrome)

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14 pages, 1125 KB

Open AccessArticle

Clinical Effectiveness, Safety, and Compliance of Two Compounded Formulations of Tacrolimus Eye Drops: An Open-Label, Sequential Prospective Study

by María Puente-Iglesias, Andrea Cuartero-Martínez, Rosario Touriño-Peralba, María Teresa Rodríguez-Ares, María Jesús Giráldez, Eva Yebra-Pimentel, Laura García-Quintanilla, Xurxo García-Otero, Miguel González-Barcia, Irene Zarra-Ferro, Francisco J. Otero-Espinar, Anxo Fernández-Ferreiro and Ana Castro-Balado

Int. J. Mol. Sci. 2024, 25(18), 9847; https://doi.org/10.3390/ijms25189847 - 12 Sep 2024

Cited by 1 | Viewed by 2410

Abstract

Ophthalmic tacrolimus compounded formulations are usually made from the commercial intravenous presentation, which contains ethanol as a solubilizer due to the low solubility of tacrolimus. The use of cyclodextrins is presented as an alternative to ethanol, an ocular irritant excipient, to avoid its long-term irritant effects. Open-label, sequential, prospective study to compare effectiveness, safety, and adherence of a new formulation of 0.015% tacrolimus with cyclodextrins (TCD) versus 0.03% tacrolimus with ethanol (TE). The ocular evaluation was assessed by ocular signs, corneal staining, subjective questionnaires as Visual Function Questionnaire (VFQ-25) and Visual Analogue Scale (VAS) of symptoms, lacrimal stability, ocular redness, and intraocular pressure. Compliance was assessed by VAS of adherence and empirically (difference between theoretical and actual consumption). Clinical ocular signs and corneal staining score remained stable for most patients 3 months after switching formulations. The TCD formulation did not modify the tear stability and intraocular pressure of the treated patients compared to the TE formulation. TCD eye drops significantly decreased the subjective pain values on VFQ-25 scale and burning sensation on the VAS symptom scale in comparison to TE formulation after 3 months after the change to TCD formulation. The novel tacrolimus in cyclodextrins formulation is a promising alternative for treating inflammatory ocular pathologies refractory to first-line treatments. Full article

(This article belongs to the Special Issue Molecular Advances in Dry Eye Syndrome)

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17 pages, 11790 KB

Open AccessArticle

Meibum Lipidomic Analysis in Evaporative Dry Eye Subjects

by Jacobo Garcia-Queiruga, Hugo Pena-Verdeal, Belen Sabucedo-Villamarin, Monica Paz-Tarrio, Esteban Guitian-Fernandez, Carlos Garcia-Resua, Eva Yebra-Pimentel and Maria J. Giraldez

Int. J. Mol. Sci. 2024, 25(9), 4782; https://doi.org/10.3390/ijms25094782 - 27 Apr 2024

Cited by 4 | Viewed by 2982

Abstract

Meibomian Glands (MG) are sebaceous glands responsible for the production of meibum, the main component of the Tear Film Lipid Layer (TFLL). The TFLL facilitates the spread of the tear film over the ocular surface, provides stability and reduces tear evaporation. Alterations in meibum composition lead to different ocular alterations like Meibomian Gland Dysfunction (MGD) and subsequent Evaporative Dry Eye (EDE). The aim of the present study was to investigate the composition and abundance of meibum lipids and their relationship with eyelid margin abnormalities, lipid layer patterns and MG status. The study utilizes a lipidomic approach to identify and quantify lipids in meibum samples using an Elute UHPLC system. This system considered all four dimensions (mass/charge, retention time, ion mobility and intensity) to provide the accurate identification of lipid species. Samples were categorized as healthy or low/no signs of alteration (group 1) or severe signs of alteration or EDE/MGD (group 2). The current investigation found differences in Variable Importance in Projection lipid abundance between both groups for the MGD signs studied. Changes in meibum composition occur and are related to higher scores in eyelid margin hyperaemia, eyelid margin irregularity, MG orifice plugging, MG loss and lipid layer pattern. Full article

(This article belongs to the Special Issue Molecular Advances in Dry Eye Syndrome)

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Review

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26 pages, 883 KB

Open AccessReview

Keratoconjunctivitis Sicca in Sjögren Disease: Diagnostic Challenges and Therapeutic Advances

by Muhammad Soyfoo, Elie Motulsky and Julie Sarrand

Int. J. Mol. Sci. 2025, 26(18), 8824; https://doi.org/10.3390/ijms26188824 - 10 Sep 2025

Viewed by 481

Abstract

Keratoconjunctivitis sicca (KCS), also commonly known as dry eye disease (DED), is one of the most prevalent and crippling features of Sjögren disease (SD), a chronic systemic autoimmune disorder featuring lymphocytic infiltration and progressive impairment of exocrine glands. KCS affects up to 95% of patients with SD and is often the earliest and most persistent manifestation, significantly compromising visual function, ocular comfort, and overall quality of life. Beyond the ocular surface, KCS mirrors a wider spectrum of immune dysregulation and epithelial damage characteristic of the disease, making it a valuable window into the underlying systemic pathology. The pathophysiology of KCS in SD is complex and multifactorial, involving an interplay between autoimmune-mediated lacrimal gland dysfunction, neuroimmune interactions, ocular surface inflammation, and epithelial instability. Tear film instability and epithelial injury result from the aberrant activation of innate and adaptive immunity, involving T and B lymphocytes, pro-inflammatory cytokines, and type I interferon pathways. Despite the clinical significance of KCS, its diagnosis remains challenging, with frequent discrepancies between subjective symptoms and objective findings. Traditional diagnostic tools often lack sensitivity and specificity, prompting the development of novel imaging techniques, tear film biomarkers, and standardized scoring systems. Concurrently, therapeutic strategies have evolved from palliative approaches to immunomodulatory and regenerative treatments, aiming to restore immune homeostasis and epithelial integrity. This review provides a comprehensive update on the pathogenesis, diagnostic landscape, and emerging treatments of KCS in the context of SD. Full article

(This article belongs to the Special Issue Molecular Advances in Dry Eye Syndrome)

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22 pages, 704 KB

Open AccessReview

Translating Biomarker Discovery: From Bench to Bedside in Dry Eye Disease

by Jeremy Jones, Kyla Frenia, Julia Gelman, Maria Beatty, Melody Zhou, Levin Ma, Sean Pieramici, Noah Eger, Deepinder Dhaliwal, Leanne T. Labriola and Kunhong Xiao

Int. J. Mol. Sci. 2025, 26(17), 8556; https://doi.org/10.3390/ijms26178556 - 3 Sep 2025

Viewed by 751

Abstract

Dry Eye Disease (DED) is a complex, multifaceted ocular disease characterized by tear film instability and inflammation. It can sometimes be elusive to identify the type of DED in patients, given the overlapping symptoms with other conditions like allergies and the multitude of stimuli that might trigger DED onset. There is also difficulty due to limitations on the diagnostic testing available to clinicians, as poor reliability and a lack of standardization plague accurate diagnoses. Identified biomarkers can help identify DED pathophysiology and category, and these include molecular biomarkers like matrix metalloproteinase-9 (MMP-9), cytokines, lactotransferrin, and lacritin, as well as functional biomarkers such as tear osmolarity. Diagnostic tools, such as the InflammaDry and I-Pen Tear Osmolarity System, also now allow for point-of-care measurement of select biomarkers, including MMP-9 and osmolarity. Nonetheless, there remains a critical need for additional, reliable, and accurate diagnostic devices to better aid in the diagnosis and management of DED. This review uniquely combines a review on the current understanding of various biomarkers with an overview of the emerging technologies available to healthcare providers, aiding in better-informed diagnosis and treatment of DED. Full article

(This article belongs to the Special Issue Molecular Advances in Dry Eye Syndrome)

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16 pages, 2970 KB

Open AccessReview

Safety and Efficacy of Diquafosol Compared to Artificial Tears for the Treatment of Dry Eye: A Systematic Review and Meta-Analysis

by José Gerardo Serrano-Robles, Ana Karen Pérez-Vázquez, Guillermo Raul Vera-Duarte, Alejandro Navas, Arturo Ramirez-Miranda, Enrique O. Graue-Hernandez and Nicolás Kahuam-López

Int. J. Mol. Sci. 2025, 26(17), 8113; https://doi.org/10.3390/ijms26178113 - 22 Aug 2025

Viewed by 1064

Abstract

Dry eye disease (DED) is a prevalent and disabling condition. Artificial tears are commonly used but often inadequate for moderate-to-severe cases. Secretagogues such as pilocarpine, cevimeline, and diquafosol offer potential alternatives, though their comparative effectiveness remains unclear. To evaluate the safety and efficacy of these secretagogues versus artificial tears in adults with DED, we searched CENTRAL, PubMed, Scopus, LILACS, ClinicalTrials.gov, and WHO ICTRP without language restrictions. Randomized controlled trials (RCTs) comparing secretagogues to artificial tears were eligible. Data extraction and synthesis were conducted using Covidence and the Cochrane RoB 2 tool, and 19 RCTs (n = 2697) were included. Fifteen were analyzed quantitatively; however, only eight trials evaluating diquafosol were suitable for meta-analysis, as data for pilocarpine and cevimeline were insufficient for quantitative synthesis. GRADE was used to assess evidence certainty. PROSPERO registration: CRD42020218407. Diquafosol significantly improved rose bengal staining at 4 weeks and OSDI scores and TBUT in post-cataract patients at 4 and 12 weeks. However, it increased mild adverse events (RR, 1.81; 95% CI, 1.15–2.84). Evidence for pilocarpine and cevimeline was limited. Diquafosol 3% shows greater efficacy than artificial tears in post-cataract DED but with more side effects. Further research is needed for other secretagogues. Full article

(This article belongs to the Special Issue Molecular Advances in Dry Eye Syndrome)

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31 pages, 1658 KB

Open AccessReview

The Role of Nerve Growth Factor on the Ocular Surface: A Review of the Current Experimental Research and Clinical Practices

by Nicolás Kahuam-López, Amir Hosseini, Jennifer Y. M. Ling, Joseph Chiang, Alfonso Iovieno and Sonia N. Yeung

Int. J. Mol. Sci. 2025, 26(13), 6012; https://doi.org/10.3390/ijms26136012 - 23 Jun 2025

Viewed by 1550

Abstract

The ocular surface is susceptible to a wide spectrum of inflammatory, degenerative, and neurotrophic diseases that can impair vision. The complex pathophysiology and limited therapeutic options associated with these conditions continue to pose significant clinical challenges. Nerve Growth Factor (NGF), a neurotrophin initially recognized for its role in neuronal survival and differentiation, has emerged as a key regulator of ocular surface homeostasis and repair. Beyond its neurotrophic functions, NGF is suggested to influence epithelial proliferation, immune responses, tear secretion, and angiogenesis. Experimental and clinical studies have implicated NGF in both the pathogenesis and potential treatment of various ocular surface diseases, including allergic conjunctivitis, neurotrophic keratopathy (NK), immune-mediated and herpetic keratitis, and dry eye disease (DED), as well as post-surgical corneal wound healing. Notably, recombinant human NGF (rhNGF, cenegermin) has been approved as the first topical biologic therapy for NK. Despite encouraging clinical outcomes, challenges such as high treatment costs, limited long-term data, and potential proangiogenic effects remain. This review consolidates current evidence on the role of NGF in ocular surface health and disease, highlighting its biological mechanisms, clinical applications, and future therapeutic potential. Full article

(This article belongs to the Special Issue Molecular Advances in Dry Eye Syndrome)

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27 pages, 2397 KB

Open AccessReview

Sex Differences in the Lacrimal Gland: Implications for Dry Eye Disease

by Snježana Kaštelan, Koraljka Hat, Zora Tomić, Tomislav Matejić and Nikola Gotovac

Int. J. Mol. Sci. 2025, 26(8), 3833; https://doi.org/10.3390/ijms26083833 - 18 Apr 2025

Viewed by 1325

Abstract

Sexual dimorphism significantly impacts the lacrimal gland’s structure, function, and ageing processes, playing an important role in dry eye disease (DED) pathophysiology. This multifactorial disorder, characterised by tear film instability, inflammation, and visual impairment, disproportionately affects women, especially after menopause. It highlights the interplay between sex steroid hormones, lacrimal gland function, and environmental factors. Systemic and local androgens are vital for maintaining lacrimal gland health and tear production, while the role of oestrogens remains less clear. Evidence suggests dose and context-dependent effects on inflammation and glandular function. Histopathological and molecular studies reveal significant sex differences in the lacrimal gland, with women exhibiting more pronounced age-related degenerative changes, including fibrosis and acinar atrophy, contributing to their increased susceptibility to DED. Despite these findings, the underlying mechanisms connecting sex steroid hormones, receptor expression, and local tissue regulation to these disparities remain poorly understood, highlighting the need for further research. This review synthesises the current knowledge of sex-specific differences in the lacrimal gland, emphasising the importance of integrating systemic and local biomarkers, histological data, and molecular insights into personalised therapeutic strategies. By tailoring treatments to patients’ unique hormonal and molecular profiles, personalised medicine has the potential to transform DED management, addressing unmet clinical needs and improving outcomes. Full article

(This article belongs to the Special Issue Molecular Advances in Dry Eye Syndrome)

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