Pub Date : 2025-04-10DOI: 10.1016/j.jtos.2025.04.005
Gary D. Novack
{"title":"Pipeline: Therapeutics – Two views of off-label use","authors":"Gary D. Novack","doi":"10.1016/j.jtos.2025.04.005","DOIUrl":"10.1016/j.jtos.2025.04.005","url":null,"abstract":"","PeriodicalId":54691,"journal":{"name":"Ocular Surface","volume":"37 ","pages":"Pages 216-218"},"PeriodicalIF":5.9,"publicationDate":"2025-04-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143834111","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-04-07DOI: 10.1016/j.jtos.2025.03.008
John K.G. Dart , Vincenzo Papa , Paolo Rama , Karl Anders Knutsson , Saj Ahmad , Scott Hau , Sara Sanchez , Antonella Franch , Federica Birattari , Pia Leon , Adriano Fasolo , Ewa Mrukwa-Kominek , Katarzyna Jadczyk-Sorek , Fiona Carley , Hossain Parwez , Darwin C. Minassian
Purpose
To compare Acanthamoeba keratitis (AK) outcomes for treatment delivered using a detailed protocol versus physician's individualised treatment.
Methods
This double cohort study compared the outcomes of these different delivery methods for PHMB 0.02 % and diamidine 0.1 % dual therapy. The primary outcome was the medical cure rate without surgery within 12 months (MCR_12) and the secondary was visual acuity. Any change of treatment, any surgery, or treatment for >12 months was a failure. Outcomes were both unadjusted and adjusted, using multivariable analysis, for baseline differences affecting outcomes. Patients were from two centres in Milan and London treated at different times; the individualised cohort (1991–2012) and per-protocol cohort (2017–2021).
Results
The individualised cohort included 96 and the per-protocol 47 patients. Both unadjusted and adjusted results were similar. The unadjusted outcomes for both centres combined showed significantly improved outcomes for per-protocol treatment with a 1.59-fold improvement in MCR_12 (95 % CI 1.40–1.80, p < 0.001) and a 2.1-fold increase in visual acuity ≥20/25 (95 % CI 1.34–3.29, p < 0.001). Amongst potential confounding factors examined, neither baseline AK disease stage, treatment centre nor the type of diamidine significantly influenced outcomes.
Conclusions
This study shows significant advantages for the use of protocol delivered versus individualised treatment for AK. The use of evidence-based treatment delivery protocols, like the one used here for AK, might improve outcomes for all causes of microbial keratitis and could offer practitioners and patients the benefit of having an easy-to-follow drug delivery protocol, with known outcomes.
目的:比较棘阿米巴角膜炎(AK)的治疗结果提供了一个详细的方案和医生的个体化治疗。方法:本双队列研究比较了0.02% PHMB和0.1%二胺双重治疗的不同给药方式的结果。主要观察指标为12个月内不手术治愈率(MCR_12),次要观察指标为视力。任何治疗方法的改变,任何手术,或长达12个月的治疗都是失败的。采用多变量分析,对影响结果的基线差异进行了未调整和调整。来自米兰和伦敦两个中心的患者在不同时间接受治疗;个体化队列(1991-2012)和按方案队列(2017-2021)。结果:个体化队列96例,按方案47例。未调整和调整后的结果相似。两个中心未经调整的合并结果显示,按方案治疗的结果显著改善,MCR_12改善1.59倍(95% CI 1.40 - 1.80, p < 0.001),视力≥20/25增加2.1倍(95% CI 1.34 - 3.29, p < 0.001)。在检查的潜在混杂因素中,基线AK疾病分期、治疗中心和二胺类型均未显著影响结果。结论:这项研究显示了使用方案传递与个性化治疗AK的显著优势。使用循证治疗方案,如这里使用的AK,可能会改善所有原因的微生物角膜炎的结果,并可以为医生和患者提供一个易于遵循的药物给药方案,已知的结果。
{"title":"Acanthamoeba keratitis treatment outcomes compared for drug delivery by protocol versus physician's individualised treatment","authors":"John K.G. Dart , Vincenzo Papa , Paolo Rama , Karl Anders Knutsson , Saj Ahmad , Scott Hau , Sara Sanchez , Antonella Franch , Federica Birattari , Pia Leon , Adriano Fasolo , Ewa Mrukwa-Kominek , Katarzyna Jadczyk-Sorek , Fiona Carley , Hossain Parwez , Darwin C. Minassian","doi":"10.1016/j.jtos.2025.03.008","DOIUrl":"10.1016/j.jtos.2025.03.008","url":null,"abstract":"<div><h3>Purpose</h3><div>To compare Acanthamoeba keratitis (AK) outcomes for treatment delivered using a detailed protocol versus physician's individualised treatment.</div></div><div><h3>Methods</h3><div>This double cohort study compared the outcomes of these different delivery methods for PHMB 0.02 % and diamidine 0.1 % dual therapy. The primary outcome was the medical cure rate without surgery within 12 months (MCR_12) and the secondary was visual acuity. Any change of treatment, any surgery, or treatment for >12 months was a failure. Outcomes were both unadjusted and adjusted, using multivariable analysis, for baseline differences affecting outcomes. Patients were from two centres in Milan and London treated at different times; the individualised cohort (1991–2012) and per-protocol cohort (2017–2021).</div></div><div><h3>Results</h3><div>The individualised cohort included 96 and the per-protocol 47 patients. Both unadjusted and adjusted results were similar. The unadjusted outcomes for both centres combined showed significantly improved outcomes for per-protocol treatment with a 1.59-fold improvement in MCR_12 (95 % CI 1.40–1.80, p < 0.001) and a 2.1-fold increase in visual acuity ≥20/25 (95 % CI 1.34–3.29, p < 0.001). Amongst potential confounding factors examined, neither baseline AK disease stage, treatment centre nor the type of diamidine significantly influenced outcomes.</div></div><div><h3>Conclusions</h3><div>This study shows significant advantages for the use of protocol delivered versus individualised treatment for AK. The use of evidence-based treatment delivery protocols, like the one used here for AK, might improve outcomes for all causes of microbial keratitis and could offer practitioners and patients the benefit of having an easy-to-follow drug delivery protocol, with known outcomes.</div></div>","PeriodicalId":54691,"journal":{"name":"Ocular Surface","volume":"38 ","pages":"Pages 132-141"},"PeriodicalIF":5.9,"publicationDate":"2025-04-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144002474","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-04-05DOI: 10.1016/j.jtos.2025.04.004
Edoardo Villani , Paolo Nucci , Jose Manuel Benitez-del-Castillo , Annegret Dahlmann-Noor , Wolf Alexander Lagrèze , Dominique Bremond-Gignac , PeDED Delphi Group
Context
Dry Eye Disease (DED) is a symptomatic multifactorial condition of the ocular surface, primarily characterized by tear film instability, which can lead to ocular surface damage. While traditionally associated with adults, recent studies have shown a significant prevalence of DED in pediatric populations, exacerbated by modern lifestyle changes such as increased screen time and environmental factors. Pediatric DED (PeDED) often goes underdiagnosed due to a lack of tailored diagnostic tools and the misattribution of symptoms to other conditions.
Methods
This Delphi study, conducted by a European panel of six ophthalmology experts sought to address expert consensus in pending challenges of PeDED.
Results
Over two rounds of surveys, 34 experts reached consensus on 39 of 46 statements, highlighting the need for age-specific diagnostic criteria and treatment protocols. The study identified environmental and lifestyle factors, such as screen time and ocular allergies, as significant risk factors for PeDED. There was strong consensus on the importance of adapting adult DED definitions and treatment approaches to better suit pediatric patients. The study also emphasized the cautious use of treatments like topical steroids or oral macrolides, especially in younger children.
Conclusion
The findings underscore the necessity for standardized clinical practices in diagnosing and managing PeDED, with a focus on improving the quality of life for affected children. Further research is required to validate these findings and develop comprehensive guidelines that cater to the unique needs of the pediatric population.
{"title":"Expert consensus on pediatric dry eye: Insights from a European Delphi study","authors":"Edoardo Villani , Paolo Nucci , Jose Manuel Benitez-del-Castillo , Annegret Dahlmann-Noor , Wolf Alexander Lagrèze , Dominique Bremond-Gignac , PeDED Delphi Group","doi":"10.1016/j.jtos.2025.04.004","DOIUrl":"10.1016/j.jtos.2025.04.004","url":null,"abstract":"<div><h3>Context</h3><div>Dry Eye Disease (DED) is a symptomatic multifactorial condition of the ocular surface, primarily characterized by tear film instability, which can lead to ocular surface damage. While traditionally associated with adults, recent studies have shown a significant prevalence of DED in pediatric populations, exacerbated by modern lifestyle changes such as increased screen time and environmental factors. Pediatric DED (PeDED) often goes underdiagnosed due to a lack of tailored diagnostic tools and the misattribution of symptoms to other conditions.</div></div><div><h3>Methods</h3><div>This Delphi study, conducted by a European panel of six ophthalmology experts sought to address expert consensus in pending challenges of PeDED.</div></div><div><h3>Results</h3><div>Over two rounds of surveys, 34 experts reached consensus on 39 of 46 statements, highlighting the need for age-specific diagnostic criteria and treatment protocols. The study identified environmental and lifestyle factors, such as screen time and ocular allergies, as significant risk factors for PeDED. There was strong consensus on the importance of adapting adult DED definitions and treatment approaches to better suit pediatric patients. The study also emphasized the cautious use of treatments like topical steroids or oral macrolides, especially in younger children.</div></div><div><h3>Conclusion</h3><div>The findings underscore the necessity for standardized clinical practices in diagnosing and managing PeDED, with a focus on improving the quality of life for affected children. Further research is required to validate these findings and develop comprehensive guidelines that cater to the unique needs of the pediatric population.</div></div>","PeriodicalId":54691,"journal":{"name":"Ocular Surface","volume":"37 ","pages":"Pages 189-197"},"PeriodicalIF":5.9,"publicationDate":"2025-04-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143799779","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
To document the expression of apical-basal polarity (ABP) determinants in the mouse corneal epithelium (CE) and elucidate the functions of Pard3 in establishment and maintenance of ABP, stratification, homeostasis, and barrier function in the CE.
Methods
Pard3Δ/ΔC mice (Pard3LoxP/LoxP; Aldh3A1-Cre/+) with cornea-specific Pard3 ablation were generated by breeding Aldh3A1-Cre/+ with Pard3LoxP/LoxP mice. The control (Aldh3A1-Cre/+ or Pard3LoxP/LoxP alone) and Pard3Δ/ΔC corneal histology, ocular surface properties, barrier function, and actin cytoskeleton were assessed by Haematoxylin and Eosin staining of paraformaldehyde-fixed, paraffin-embedded tissues, scanning electron microscopy, fluorescein staining, and phalloidin staining, respectively. The expression of specific markers of interest was evaluated by qRT-PCR, immunoblots and immunofluorescent staining.
Results
Dynamic changes were observed in the expression and localization of ABP determinants as the CE stratified and matured between post-natal day 5 (PN5) and PN52. Adult Pard3Δ/ΔC CE contained fewer cell layers with rounded basal cells, and loosely adherent superficial cells lacking microplicae. Adult Pard3Δ/ΔC CE also displayed impaired barrier function with decreased expression of tight junction, adherens junction, and desmosome components, disrupted actin cytoskeletal organization, increased proliferation, and upregulation of transcription factors that drive epithelial-mesenchymal transition (EMT).
Conclusions
Disruption of ABP in Pard3Δ/ΔC CE, altered expression of cell junction complex components and disorganized actin cytoskeleton, increased cell proliferation, and upregulated EMT transcription factors suggest that the ABP-determinant Pard3 promotes CE features while suppressing mesenchymal cell fate. Collectively, these results elucidate that Pard3-mediated ABP is essential for CE stratification, homeostasis and barrier function.
{"title":"Pard3 promotes corneal epithelial stratification and homeostasis by regulating apical-basal polarity, cytoskeletal organization and tight junction-mediated barrier function","authors":"Mehak Vohra , Simran Kumar , Peri Sohnen , Satinder Kaur , Sudha Swamynathan , Tomonori Hirose , Zbynek Kozmik , Shivalingappa K. Swamynathan","doi":"10.1016/j.jtos.2025.04.001","DOIUrl":"10.1016/j.jtos.2025.04.001","url":null,"abstract":"<div><h3>Purpose</h3><div>To document the expression of apical-basal polarity (ABP) determinants in the mouse corneal epithelium (CE) and elucidate the functions of Pard3 in establishment and maintenance of ABP, stratification, homeostasis, and barrier function in the CE.</div></div><div><h3>Methods</h3><div><em>Pard3</em><sup>Δ/ΔC</sup> mice (<em>Pard3</em><sup><em>LoxP/LoxP</em></sup>; <em>Aldh3A1-Cre/+</em>) with cornea-specific <em>Pard3</em> ablation were generated by breeding <em>Aldh3A1-Cre/+</em> with <em>Pard3</em><sup><em>LoxP/LoxP</em></sup> mice. The control (<em>Aldh3A1-Cre/+</em> or <em>Pard3</em><sup><em>LoxP/LoxP</em></sup> alone) and <em>Pard3</em><sup>Δ/ΔC</sup> corneal histology, ocular surface properties, barrier function, and actin cytoskeleton were assessed by Haematoxylin and Eosin staining of paraformaldehyde-fixed, paraffin-embedded tissues, scanning electron microscopy, fluorescein staining, and phalloidin staining, respectively. The expression of specific markers of interest was evaluated by qRT-PCR, immunoblots and immunofluorescent staining.</div></div><div><h3>Results</h3><div>Dynamic changes were observed in the expression and localization of ABP determinants as the CE stratified and matured between post-natal day 5 (PN5) and PN52. Adult <em>Pard3</em><sup>Δ/ΔC</sup> CE contained fewer cell layers with rounded basal cells, and loosely adherent superficial cells lacking microplicae. Adult <em>Pard3</em><sup>Δ/ΔC</sup> CE also displayed impaired barrier function with decreased expression of tight junction, adherens junction, and desmosome components, disrupted actin cytoskeletal organization, increased proliferation, and upregulation of transcription factors that drive epithelial-mesenchymal transition (EMT).</div></div><div><h3>Conclusions</h3><div>Disruption of ABP in <em>Pard3</em><sup>Δ/ΔC</sup> CE, altered expression of cell junction complex components and disorganized actin cytoskeleton, increased cell proliferation, and upregulated EMT transcription factors suggest that the ABP-determinant Pard3 promotes CE features while suppressing mesenchymal cell fate. Collectively, these results elucidate that Pard3-mediated ABP is essential for CE stratification, homeostasis and barrier function.</div></div>","PeriodicalId":54691,"journal":{"name":"Ocular Surface","volume":"37 ","pages":"Pages 201-215"},"PeriodicalIF":5.9,"publicationDate":"2025-04-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143797367","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Bacteria are the leading cause of ocular infections (OIs), accounting for 32 %–74 % of cases globally. Coagulase-negative Staphylococci (CoNS) are frequently identified as the main causative pathogens of all types of OIs. This study systematically reviewed the prevalence of CoNS and their resistance rates to ciprofloxacin, chloramphenicol, tetracycline, and trimethoprim/sulfamethoxazole in OIs worldwide.
We conducted a comprehensive search of publications from January 1, 2000, to May 19, 2024, using three major databases: Scopus, Web of Science, and PubMed. Statistical analyses were performed with Stata 14.0, employing the Metaprop package to calculate pooled prevalence and confidence intervals. A random-effects model with double arcsine transformations stabilized variance in meta-analyses, and publication bias was evaluated using the “metabias” command. If bias was detected, we adjusted prevalence using the trim-and-fill method. A significant level of 0.05 was set for all analyses.
Out of 2116 articles, 214 were eligible for data extraction. The pooled prevalence of CoNS isolates in OIs was 25 % (95 % CI: 21–28; I2 = 99.7 %), with Europe showing the highest prevalence at 30 % (95 % CI: 15–46; I2: 99.9 %) and Asia the lowest at 21 % (95 % CI: 19–24; I2: 98.68 %). The pooled prevalence of resistance among CoNS isolates from patients with OIs was 27 % (95 % CI: 19–35) for ciprofloxacin, 23 % (95 % CI: 12–34) for chloramphenicol, 41 % (95 % CI: 33–49) for tetracycline, and 40 % (95 % CI: 29–51) for trimethoprim/sulfamethoxazole.
In conclusion, CoNS are significant pathogens in OIs, highlighting the need for improved diagnostic tests and updated management guidelines globally.
{"title":"Prevalence and antimicrobial susceptibility pattern of coagulase-negative Staphylococci isolated from ocular infections: A systematic review and meta-analysis","authors":"Abolfazl Rafati Zomorodi , Maryam Mohammadi , Zahra Gholizadeh Farshi , Mohammad Hassan Parvizi Mashhadi , Sasan Pourbagher Benam , Mohammad Motamedifar","doi":"10.1016/j.jtos.2025.03.006","DOIUrl":"10.1016/j.jtos.2025.03.006","url":null,"abstract":"<div><div>Bacteria are the leading cause of ocular infections (OIs), accounting for 32 %–74 % of cases globally. Coagulase-negative <em>Staphylococci</em> (CoNS) are frequently identified as the main causative pathogens of all types of OIs. This study systematically reviewed the prevalence of CoNS and their resistance rates to ciprofloxacin, chloramphenicol, tetracycline, and trimethoprim/sulfamethoxazole in OIs worldwide.</div><div>We conducted a comprehensive search of publications from January 1, 2000, to May 19, 2024, using three major databases: Scopus, Web of Science, and PubMed. Statistical analyses were performed with Stata 14.0, employing the Metaprop package to calculate pooled prevalence and confidence intervals. A random-effects model with double arcsine transformations stabilized variance in meta-analyses, and publication bias was evaluated using the “metabias” command. If bias was detected, we adjusted prevalence using the trim-and-fill method. A significant level of 0.05 was set for all analyses.</div><div>Out of 2116 articles, 214 were eligible for data extraction. The pooled prevalence of CoNS isolates in OIs was 25 % (95 % CI: 21–28; I<sup>2</sup> = 99.7 %), with Europe showing the highest prevalence at 30 % (95 % CI: 15–46; I<sup>2</sup>: 99.9 %) and Asia the lowest at 21 % (95 % CI: 19–24; I<sup>2</sup>: 98.68 %). The pooled prevalence of resistance among CoNS isolates from patients with OIs was 27 % (95 % CI: 19–35) for ciprofloxacin, 23 % (95 % CI: 12–34) for chloramphenicol, 41 % (95 % CI: 33–49) for tetracycline, and 40 % (95 % CI: 29–51) for trimethoprim/sulfamethoxazole.</div><div>In conclusion, CoNS are significant pathogens in OIs, highlighting the need for improved diagnostic tests and updated management guidelines globally.</div></div>","PeriodicalId":54691,"journal":{"name":"Ocular Surface","volume":"37 ","pages":"Pages 227-235"},"PeriodicalIF":5.9,"publicationDate":"2025-04-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143797369","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-04-02DOI: 10.1016/j.jtos.2025.03.009
Michael H. Goldstein , Deepinder Dhaliwal , Ed Isbey , Stephen Pflugfelder , Mark J. Mannis , Gary D. Novack
{"title":"A tribute to Gary N. Foulks, M.D. (1944–2025) and his contributions to the ocular surface","authors":"Michael H. Goldstein , Deepinder Dhaliwal , Ed Isbey , Stephen Pflugfelder , Mark J. Mannis , Gary D. Novack","doi":"10.1016/j.jtos.2025.03.009","DOIUrl":"10.1016/j.jtos.2025.03.009","url":null,"abstract":"","PeriodicalId":54691,"journal":{"name":"Ocular Surface","volume":"37 ","pages":"Pages 198-200"},"PeriodicalIF":5.9,"publicationDate":"2025-04-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143789464","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-04-01Epub Date: 2025-01-31DOI: 10.1016/j.jtos.2025.01.013
Yong Yuan , Shingo Yasuda , Kaitlyn L. Funk , Winston Kao , Shizuya Saika , Adam Kaufman , Chia-Yang Liu
Purpose
To understand how Tbr1 and Smad4 play a pivotal role in controlling ECM synthesis versus degradation for maintaining corneal stromal homeostasis and otherwise leading to corneal ectasia.
Methods
Keratocyte-specific and inducible knockout (iKO) of Tbr1, Smad4, or Tbr1/Smad4 double KO (iDKO) mice were generated. OCT was used to assess corneal thickness in vivo. Masson's trichrome and collagen hybridizing peptide stainings were performed to examine collagen expression. Immunostaining with an anti-cathepsin B antibody was used to assess ECM degradation. Cathepsin B inhibitor, CA-074Me, eyedrop was conducted to test its effect on treating stromal thinning in Tbr1 iKO mice.
Results
Tbr1 iKO and Smad4 iKO displayed corneal thinning, but Tbr1 iKO revealed a progressive and more severe pathology than Smad4 iKO. Tbr1 iKO cornea lost most of its stroma and thus a dome shape. Collagen ECM is evenly distributed in Smad4 iKO as well as control littermates but was lost mainly in the anterior stroma of the Tbr1 iKO. Interestingly, Tbr1/Smad4 iDKO ameliorated Tbr1 iKO phenotype. The basal level of Cathepsin b (Ctsb) could be detected in the control stroma but was significantly increased in the Tbr1 iKO stromal cells and this effect was canceled in Tbr1/Smad4 iDKO. CA-074Me eyedrops administration significantly inhibited progressive corneal thinning caused by the Tbr1 iKO.
Conclusion
Our data from Tbr1/Smad4 iDKO argued that Smad4 played a pivotal role in controlling Tbr1-dependent ECM synthesis and Tbr1-independent ECM degradation to maintain corneal stromal integrity and homeostasis.
{"title":"Smad4 deficiency ameliorates the progressive corneal stroma thinning caused by the loss of Tbr1","authors":"Yong Yuan , Shingo Yasuda , Kaitlyn L. Funk , Winston Kao , Shizuya Saika , Adam Kaufman , Chia-Yang Liu","doi":"10.1016/j.jtos.2025.01.013","DOIUrl":"10.1016/j.jtos.2025.01.013","url":null,"abstract":"<div><h3>Purpose</h3><div>To understand how <em>Tbr1</em> and <em>Smad4</em> play a pivotal role in controlling ECM synthesis versus degradation for maintaining corneal stromal homeostasis and otherwise leading to corneal ectasia.</div></div><div><h3>Methods</h3><div>Keratocyte-specific and inducible knockout (iKO) of <em>Tbr1</em>, <em>Smad4</em>, or <em>Tbr1/Smad4</em> double KO (iDKO) mice were generated. OCT was used to assess corneal thickness <em>in vivo</em>. Masson's trichrome and collagen hybridizing peptide stainings were performed to examine collagen expression. Immunostaining with an anti-cathepsin B antibody was used to assess ECM degradation. Cathepsin B inhibitor, CA-074Me, eyedrop was conducted to test its effect on treating stromal thinning in <em>Tbr1</em> iKO mice.</div></div><div><h3>Results</h3><div><em>Tbr1</em> iKO and <em>Smad4</em> iKO displayed corneal thinning, but <em>Tbr1</em> iKO revealed a progressive and more severe pathology than <em>Smad4</em> iKO. <em>Tbr1</em> iKO cornea lost most of its stroma and thus a dome shape. Collagen ECM is evenly distributed in <em>Smad4</em> iKO as well as control littermates but was lost mainly in the anterior stroma of the <em>Tbr1</em> iKO. Interestingly, <em>Tbr1/Smad4</em> iDKO ameliorated <em>Tbr1</em> iKO phenotype. The basal level of Cathepsin b (Ctsb) could be detected in the control stroma but was significantly increased in the <em>Tbr1</em> iKO stromal cells and this effect was canceled in <em>Tbr1/Smad4</em> iDKO. CA-074Me eyedrops administration significantly inhibited progressive corneal thinning caused by the <em>Tbr1</em> iKO.</div></div><div><h3>Conclusion</h3><div>Our data from <em>Tbr1/Smad4</em> iDKO argued that Smad4 played a pivotal role in controlling Tbr1-dependent ECM synthesis and Tbr1-independent ECM degradation to maintain corneal stromal integrity and homeostasis.</div></div>","PeriodicalId":54691,"journal":{"name":"Ocular Surface","volume":"36 ","pages":"Pages 181-189"},"PeriodicalIF":5.9,"publicationDate":"2025-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143082811","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-04-01DOI: 10.1016/j.jtos.2025.03.007
Sonali Pal-Ghosh , Himani Datta-Majumdar , Soneha Datta , Shelly Dimri , Jordan Hally , Hugo Wehmeyer , Zhong Chen , Mitchell Watsky , Jian-Xing Ma , Wentao Liang , Mary Ann Stepp
Purpose
To determine the mechanisms used to internalize mitochondria by corneal epithelial cells after in vivo corneal trephine injury and in vitro in corneal epithelial cells.
Methods
Male and female mice were subjected to trephine injury and euthanized immediately, 6, and 24 h after injury. Macropinocytosis was quantified in vivo using 70 kD fluorescent dextran. Mitochondrial content was assessed by immunofluorescence and metabolic activity quantified by Seahorse assay immediately and 6 h after injury. In vitro experiments using human corneal and limbal epithelial (HCLE) cells and isolated mitochondria were performed to assess mitochondrial transfer in the presence of the gap junction inhibitor 18α-glycyrrhetinc acid and the macropincytosis inhibitor ethylisopropylamiloride.
Results
Mitochondria accumulate within apical epithelial cell layers within minutes of trephine injury. Macropinocytosis also increases within minutes of trephine injury. Oxygen Consumption Rates increase in the corneal epithelium 6 h after trephine injury in males and females. Inhibiting gap junctions increases mitochondrial engulfment while inhibiting macropinocytosis prevents engulfment of mitochondria by corneal epithelial cells in vitro.
Conclusions
Molecules released by injured cells and severed axons induce macropinocytosis in corneal epithelial cells within minutes of trephine injury. An increase in oxygen consumption rate in the corneal epithelium after trephine injury indicates that axonal mitochondria can evade lysosomal degradation for at least 6 h. In vitro studies using isolated labeled and unlabeled mitochondria and control and mechanically stressed human corneal epithelial cells confirm the involvement of macropinocytosis in the engulfment of free and vesicle bound mitochondria by corneal epithelial cells.
{"title":"Corneal epithelial cells upregulate macropinocytosis to engulf metabolically active axonal mitochondria released by injured axons","authors":"Sonali Pal-Ghosh , Himani Datta-Majumdar , Soneha Datta , Shelly Dimri , Jordan Hally , Hugo Wehmeyer , Zhong Chen , Mitchell Watsky , Jian-Xing Ma , Wentao Liang , Mary Ann Stepp","doi":"10.1016/j.jtos.2025.03.007","DOIUrl":"10.1016/j.jtos.2025.03.007","url":null,"abstract":"<div><h3>Purpose</h3><div>To determine the mechanisms used to internalize mitochondria by corneal epithelial cells after <em>in vivo</em> corneal trephine injury and <em>in vitro</em> in corneal epithelial cells.</div></div><div><h3>Methods</h3><div>Male and female mice were subjected to trephine injury and euthanized immediately, 6, and 24 h after injury. Macropinocytosis was quantified <em>in vivo</em> using 70 kD fluorescent dextran. Mitochondrial content was assessed by immunofluorescence and metabolic activity quantified by Seahorse assay immediately and 6 h after injury. <em>In vitro</em> experiments using human corneal and limbal epithelial (HCLE) cells and isolated mitochondria were performed to assess mitochondrial transfer in the presence of the gap junction inhibitor 18α-glycyrrhetinc acid and the macropincytosis inhibitor ethylisopropylamiloride.</div></div><div><h3>Results</h3><div>Mitochondria accumulate within apical epithelial cell layers within minutes of trephine injury. Macropinocytosis also increases within minutes of trephine injury. Oxygen Consumption Rates increase in the corneal epithelium 6 h after trephine injury in males and females. Inhibiting gap junctions increases mitochondrial engulfment while inhibiting macropinocytosis prevents engulfment of mitochondria by corneal epithelial cells <em>in vitro</em>.</div></div><div><h3>Conclusions</h3><div>Molecules released by injured cells and severed axons induce macropinocytosis in corneal epithelial cells within minutes of trephine injury. An increase in oxygen consumption rate in the corneal epithelium after trephine injury indicates that axonal mitochondria can evade lysosomal degradation for at least 6 h. <em>In vitro</em> studies using isolated labeled and unlabeled mitochondria and control and mechanically stressed human corneal epithelial cells confirm the involvement of macropinocytosis in the engulfment of free and vesicle bound mitochondria by corneal epithelial cells.</div></div>","PeriodicalId":54691,"journal":{"name":"Ocular Surface","volume":"37 ","pages":"Pages 173-188"},"PeriodicalIF":5.9,"publicationDate":"2025-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143777254","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
To ascertain the homing of monocytes and neutrophils in the epithelium versus stroma of HSV-1 infected corneas at different stages of infection and functional significance of their anatomical location in virus-infected corneas.
Methods
The corneas of C57BL/6J mice were infected with HSV-1 McKrae. Mice were euthanized on different days post-infection. The epithelium and stroma were separated from the infected corneas, and flow cytometry was performed to characterize the myeloid cell subsets in the epithelium versus the stromal layers of an infected cornea. MACS columns were used to purify neutrophils or deplete myeloid cells from infected corneas. Corneal epithelial scratch assay was performed to ascertain the impact of neutrophils on epithelium wound healing.
Results
Our results showed a biphasic influx of monocytes in the epithelial but not the stromal layer of HSV-1-infected corneas. Furthermore, we noted the predominance of monocytes over neutrophils in the epithelium and the stromal layer of the cornea during the pre-clinical stage of corneal HSV-1 infection. However, neutrophils were the major myeloid cell subset in the epithelium and stroma during the clinical disease period of infection. Removal of monocytes from the infected epithelial layer during the pre-clinical stage promotes the dissemination of the virus. Interestingly, neutrophils localized in the corneal epithelium inhibit corneal epithelial wound healing.
Conclusions
Together, our data suggest that differential kinetics of monocytes and neutrophils homing in the epithelial layer regulate viral dissemination and epithelial wound healing in HSV-1-infected corneas.
{"title":"Differential homing of monocytes and neutrophils in the epithelial layer of HSV-1 infected cornea regulates viral dissemination and wound healing","authors":"Mizumi Setia, Pratima Krishna Suvas, Mashidur Rana, Anish Chakraborty, Susmit Suvas","doi":"10.1016/j.jtos.2025.01.002","DOIUrl":"10.1016/j.jtos.2025.01.002","url":null,"abstract":"<div><h3>Purpose</h3><div>To ascertain the homing of monocytes and neutrophils in the epithelium versus stroma of HSV-1 infected corneas at different stages of infection and functional significance of their anatomical location in virus-infected corneas.</div></div><div><h3>Methods</h3><div>The corneas of C57BL/6J mice were infected with HSV-1 McKrae. Mice were euthanized on different days post-infection. The epithelium and stroma were separated from the infected corneas, and flow cytometry was performed to characterize the myeloid cell subsets in the epithelium versus the stromal layers of an infected cornea. MACS columns were used to purify neutrophils or deplete myeloid cells from infected corneas. Corneal epithelial scratch assay was performed to ascertain the impact of neutrophils on epithelium wound healing.</div></div><div><h3>Results</h3><div>Our results showed a biphasic influx of monocytes in the epithelial but not the stromal layer of HSV-1-infected corneas. Furthermore, we noted the predominance of monocytes over neutrophils in the epithelium and the stromal layer of the cornea during the pre-clinical stage of corneal HSV-1 infection. However, neutrophils were the major myeloid cell subset in the epithelium and stroma during the clinical disease period of infection. Removal of monocytes from the infected epithelial layer during the pre-clinical stage promotes the dissemination of the virus. Interestingly, neutrophils localized in the corneal epithelium inhibit corneal epithelial wound healing.</div></div><div><h3>Conclusions</h3><div>Together, our data suggest that differential kinetics of monocytes and neutrophils homing in the epithelial layer regulate viral dissemination and epithelial wound healing in HSV-1-infected corneas.</div></div>","PeriodicalId":54691,"journal":{"name":"Ocular Surface","volume":"36 ","pages":"Pages 69-82"},"PeriodicalIF":5.9,"publicationDate":"2025-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142967683","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-04-01Epub Date: 2025-01-31DOI: 10.1016/j.jtos.2025.01.015
Fiona Stapleton , Tianni Jia , Venita DePuy , Charles Bosworth , Marc Gleeson , Jacqueline Tan
Purpose
To explore effects of topical 1 % selenium sulfide on signs and symptoms in symptomatic contact lens-wearers, in an exploratory 4-month prospective placebo-controlled double-masked randomised trial.
Methods
Symptomatic wearers (Contact Lens Dry Eye Questionnaire-8 [CLDEQ-8] score>12) with meibomian gland dysfunction (meibomian gland score (MGS)≤12), were enrolled and received either active (AZR-MD-001-containing 1 % selenium sulfide), or vehicle ointment, to the lower eyelid margin twice-weekly. MGS, meibomian glands-yielding liquid secretion (MGYLS), lipid layer thickness, tear meniscus height, tear break-up time, tear evaporation rate, lid wiper epitheliopathy, CLDEQ-8 and comfortable wear time (CWT) were measured at baseline and to 4-months. Differences between active and placebo were compared to baseline.
Results
Fourteen participants (5M:9 F, 30.8 ± 13.8 years) completed the study. In the active group, change in MGS from baseline improved by 1-month (mean difference 7.9 ± 8.0, p = 0.03), to 4-months (16.0 ± 11.3, p < 0.01). MGYLS improved from baseline by 1.5-months (4.0 ± 3.3) to 4-months (4.1 ± 4.3, p < 0.01). In the vehicle, change in MGS (12.1 ± 10.7) and MGYLS (3.9 ± 3.2) were improved at 4-months only (p < 0.01). CLDEQ-8 score improved at 1-month and 4-months compared to baseline (−4.4 ± 3.2, −5.1 ± 4.7, p ≤ 0.02) in the active and at 4-months only in the vehicle group (−4.4 ± 6.4, p = 0.02). In the active group, CLDEQ-8 visual function scores improved at 1- and 4-months (p ≤ 0.02) and CWT at 4-months (median 7 vs.10 h, p = 0.025). Other signs were unchanged.
Conclusions
This exploratory study indicates that twice-weekly use of AZR-MD-001 ointment can rapidly improve gland patency and secretion in symptomatic contact lens-wearers. AZR-MD-001 reduced changeable/blurry vision and prolonged CWT, suggesting relevant future endpoints.
{"title":"The effect of a biweekly novel selenium sulfide-containing topical treatment in symptomatic contact lens wearers: An exploratory study","authors":"Fiona Stapleton , Tianni Jia , Venita DePuy , Charles Bosworth , Marc Gleeson , Jacqueline Tan","doi":"10.1016/j.jtos.2025.01.015","DOIUrl":"10.1016/j.jtos.2025.01.015","url":null,"abstract":"<div><h3>Purpose</h3><div>To explore effects of topical 1 % selenium sulfide on signs and symptoms in symptomatic contact lens-wearers, in an exploratory 4-month prospective placebo-controlled double-masked randomised trial.</div></div><div><h3>Methods</h3><div>Symptomatic wearers (Contact Lens Dry Eye Questionnaire-8 [CLDEQ-8] score>12) with meibomian gland dysfunction (meibomian gland score (MGS)≤12), were enrolled and received either active (AZR-MD-001-containing 1 % selenium sulfide), or vehicle ointment, to the lower eyelid margin twice-weekly. MGS, meibomian glands-yielding liquid secretion (MGYLS), lipid layer thickness, tear meniscus height, tear break-up time, tear evaporation rate, lid wiper epitheliopathy, CLDEQ-8 and comfortable wear time (CWT) were measured at baseline and to 4-months. Differences between active and placebo were compared to baseline.</div></div><div><h3>Results</h3><div>Fourteen participants (5M:9 F, 30.8 ± 13.8 years) completed the study. In the active group, change in MGS from baseline improved by 1-month (mean difference 7.9 ± 8.0, p = 0.03), to 4-months (16.0 ± 11.3, p < 0.01). MGYLS improved from baseline by 1.5-months (4.0 ± 3.3) to 4-months (4.1 ± 4.3, p < 0.01). In the vehicle, change in MGS (12.1 ± 10.7) and MGYLS (3.9 ± 3.2) were improved at 4-months only (p < 0.01). CLDEQ-8 score improved at 1-month and 4-months compared to baseline (−4.4 ± 3.2, −5.1 ± 4.7, p ≤ 0.02) in the active and at 4-months only in the vehicle group (−4.4 ± 6.4, p = 0.02). In the active group, CLDEQ-8 visual function scores improved at 1- and 4-months (p ≤ 0.02) and CWT at 4-months (median 7 vs.10 h, p = 0.025). Other signs were unchanged.</div></div><div><h3>Conclusions</h3><div>This exploratory study indicates that twice-weekly use of AZR-MD-001 ointment can rapidly improve gland patency and secretion in symptomatic contact lens-wearers. AZR-MD-001 reduced changeable/blurry vision and prolonged CWT, suggesting relevant future endpoints.</div></div>","PeriodicalId":54691,"journal":{"name":"Ocular Surface","volume":"36 ","pages":"Pages 190-197"},"PeriodicalIF":5.9,"publicationDate":"2025-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143082778","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}