Experimental eye research最新文献

TCF4 expansion–associated loss of FN1 intron retention drives extracellular matrix accumulation in Fuchs endothelial corneal dystrophy

IF 3 2区医学 Q1 OPHTHALMOLOGY

Experimental eye research

Pub Date : 2025-04-21 DOI: 10.1016/j.exer.2025.110398

Soichiro Inagaki , Taichi Yuasa , Theofilos Tourtas , Ursula Schlötzer-Schrehardt , Friedrich Kruse , Noriko Koizumi , Naoki Okumura

Fuchs endothelial corneal dystrophy (FECD), which is characterized by excessive extracellular matrix (ECM) accumulation and corneal endothelial cell degeneration, has trinucleotide repeat expansion in TCF4 as a major genetic risk factor. While aberrant splicing has been implicated in FECD pathogenesis, the mechanistic link between splicing abnormalities and disease-specific features remains unclear. Here, we investigated the intron retention (IR) patterns in corneal endothelial cells from FECD patients with TCF4 expansion. Initial RNA-Seq analysis using rMATS identified 486 upregulated and 89 downregulated IR events in expansion-positive FECD compared to controls. Subsequent analysis with the more stringent IRFinder algorithm revealed 10 upregulated IR events distributed across nine genes and, notably, 6 downregulated events exclusively localized within FN1, a major component of corneal guttae. While DEXSeq analysis showed reduced expression across FN1 gene regions in FECD samples, subsequent qPCR validation in an independent cohort demonstrated significantly elevated FN1 expression in both expansion-positive and expansion-negative FECD samples compared to controls. This paradoxical finding suggests that the loss of normal IR-mediated regulation may contribute to pathological FN1 overexpression in FECD. Gene ontology analysis of IR-associated genes revealed enrichment in RNA splicing and ECM-related pathways, supporting a role for IR in disease pathogenesis. Our findings reveal an association between TCF4 expansion and reduced FN1 intron retention, which correlates with ECM accumulation, suggesting a potential link between RNA processing alterations and hallmark features of FECD. These results suggest that targeting IR-mediated regulation could represent a therapeutic strategy for preventing disease progression.

{"title":"TCF4 expansion–associated loss of FN1 intron retention drives extracellular matrix accumulation in Fuchs endothelial corneal dystrophy","authors":"Soichiro Inagaki , Taichi Yuasa , Theofilos Tourtas , Ursula Schlötzer-Schrehardt , Friedrich Kruse , Noriko Koizumi , Naoki Okumura","doi":"10.1016/j.exer.2025.110398","DOIUrl":"10.1016/j.exer.2025.110398","url":null,"abstract":"<div><div>Fuchs endothelial corneal dystrophy (FECD), which is characterized by excessive extracellular matrix (ECM) accumulation and corneal endothelial cell degeneration, has trinucleotide repeat expansion in <em>TCF4</em> as a major genetic risk factor. While aberrant splicing has been implicated in FECD pathogenesis, the mechanistic link between splicing abnormalities and disease-specific features remains unclear. Here, we investigated the intron retention (IR) patterns in corneal endothelial cells from FECD patients with <em>TCF4</em> expansion. Initial RNA-Seq analysis using rMATS identified 486 upregulated and 89 downregulated IR events in expansion-positive FECD compared to controls. Subsequent analysis with the more stringent IRFinder algorithm revealed 10 upregulated IR events distributed across nine genes and, notably, 6 downregulated events exclusively localized within <em>FN1</em>, a major component of corneal guttae. While DEXSeq analysis showed reduced expression across <em>FN1</em> gene regions in FECD samples, subsequent qPCR validation in an independent cohort demonstrated significantly elevated <em>FN1</em> expression in both expansion-positive and expansion-negative FECD samples compared to controls. This paradoxical finding suggests that the loss of normal IR-mediated regulation may contribute to pathological <em>FN1</em> overexpression in FECD. Gene ontology analysis of IR-associated genes revealed enrichment in RNA splicing and ECM-related pathways, supporting a role for IR in disease pathogenesis. Our findings reveal an association between <em>TCF4</em> expansion and reduced <em>FN1</em> intron retention, which correlates with ECM accumulation, suggesting a potential link between RNA processing alterations and hallmark features of FECD. These results suggest that targeting IR-mediated regulation could represent a therapeutic strategy for preventing disease progression.</div></div>","PeriodicalId":12177,"journal":{"name":"Experimental eye research","volume":"255 ","pages":"Article 110398"},"PeriodicalIF":3.0,"publicationDate":"2025-04-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143859824","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Reconstruction of highly and extremely aberrated wavefront for ocular Shack-Hartmann sensor using multi-task Attention-UNet

IF 3 2区医学 Q1 OPHTHALMOLOGY

Experimental eye research

Pub Date : 2025-04-18 DOI: 10.1016/j.exer.2025.110394

Yibin Tian , Zipei Luo , Dajiang Lu , Cheng Liu , Christine Wildsoet

In certain ocular conditions, such as in eyes with keratoconus or after corneal laser surgery, Higher Order Aberrations (HOAs) may be dramatically elevated. Accurately recording interpretable wavefronts in such highly aberrated eyes using Shack-Hartmann sensor is a challenging task. While there are studies that have applied deep neural networks to Shack-Hartmann wavefront reconstructions, they have been limited to low resolution and small dynamic range cases. In this study, we introduce a multi-task learning scheme for High-Resolution and High Dynamic Range Shack-Hartmann wavefront reconstruction using a modified attention-UNet (HR-HDR-SHUNet), which outputs a wavefront map along with Zernike coefficients simultaneously. The HR-HDR-SHUNet was evaluated on three large datasets with different levels of HOAs (regularly, highly, and extremely aberrated), with successful reconstruction of all aberrated wavefronts, at the same time achieving significantly higher accuracy than both traditional methods and other deep learning networks; it is also computationally more efficient than the latter.

{"title":"Reconstruction of highly and extremely aberrated wavefront for ocular Shack-Hartmann sensor using multi-task Attention-UNet","authors":"Yibin Tian , Zipei Luo , Dajiang Lu , Cheng Liu , Christine Wildsoet","doi":"10.1016/j.exer.2025.110394","DOIUrl":"10.1016/j.exer.2025.110394","url":null,"abstract":"<div><div>In certain ocular conditions, such as in eyes with keratoconus or after corneal laser surgery, Higher Order Aberrations (HOAs) may be dramatically elevated. Accurately recording interpretable wavefronts in such highly aberrated eyes using Shack-Hartmann sensor is a challenging task. While there are studies that have applied deep neural networks to Shack-Hartmann wavefront reconstructions, they have been limited to low resolution and small dynamic range cases. In this study, we introduce a multi-task learning scheme for High-Resolution and High Dynamic Range Shack-Hartmann wavefront reconstruction using a modified attention-UNet (HR-HDR-SHUNet), which outputs a wavefront map along with Zernike coefficients simultaneously. The HR-HDR-SHUNet was evaluated on three large datasets with different levels of HOAs (regularly, highly, and extremely aberrated), with successful reconstruction of all aberrated wavefronts, at the same time achieving significantly higher accuracy than both traditional methods and other deep learning networks; it is also computationally more efficient than the latter.</div></div>","PeriodicalId":12177,"journal":{"name":"Experimental eye research","volume":"255 ","pages":"Article 110394"},"PeriodicalIF":3.0,"publicationDate":"2025-04-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143870473","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

APEX1 attenuates ERS-induced paraptosis by inhibiting the P53 pathway in LECs

IF 3 2区医学 Q1 OPHTHALMOLOGY

Experimental eye research

Pub Date : 2025-04-18 DOI: 10.1016/j.exer.2025.110393

Renhao Zhong , Lihua Kang , Wenjing Geng, Linhui Xu, Pengfei Li, Miaomiao Wu, Guowei Zhang, Mengying Zhou, Kai Zhang, Min Ji, Huaijin Guan

Age-related cortical cataract (ARCC) is a prominent subtype of cataract, characterized by the presence of vacuoles and spoke-like opacity. Previous studies have suggested that paraptosis is involved in the onset of early ARCC vacuolar degeneration. In this experiment, hydrogen peroxide (H₂O₂)-induced SRA01/04 cells were used to establish a paraptosis-like cell model, and the function and underlying mechanism of APEX1 in this cell model were explored. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) and Western Blot analyses were conducted to assess the expression of pertinent genes in SRA01/04. Confocal fluorescence microscopy, using ER-tracker kits, was applied to clarify the relationship between the endoplasmic reticulum and intracellular vacuoles. The co-IP assay was used to verify the interaction between APEX1 and P53. The GTRD database was employed to predict the putative target genes combined with P53, and CUT&RUN assay was employed to confirm the enrichment of the P53 and ATF6 promoters following APEX1 overexpression. Firstly, the pathological sections of the vacuolar degeneration zone in the lens cortex of ARCC patients exhibited fiber disarray and vacuole development. Meanwhile, the protein expression of Alix, a specific paraptosis inhibitor, was decreased in low-concentration H₂O₂-treated SRA01/04 cells. Secondly, we discovered that 4-PBA suppressed the expression of ATF6 and PERK. Moreover, overexpression of APEX1 in SRA01/04 cells improved endoplasmic reticulum morphology, inhibited the interaction between P53 and ATF6, and attenuated paraptosis in SRA01/04. APEX1 regulated P53 and then mediated ATF6 to affect the endoplasmic reticulum stress and paraptosis in H₂O₂-induced SRA01/04 cells.

{"title":"APEX1 attenuates ERS-induced paraptosis by inhibiting the P53 pathway in LECs","authors":"Renhao Zhong , Lihua Kang , Wenjing Geng, Linhui Xu, Pengfei Li, Miaomiao Wu, Guowei Zhang, Mengying Zhou, Kai Zhang, Min Ji, Huaijin Guan","doi":"10.1016/j.exer.2025.110393","DOIUrl":"10.1016/j.exer.2025.110393","url":null,"abstract":"<div><div>Age-related cortical cataract (ARCC) is a prominent subtype of cataract, characterized by the presence of vacuoles and spoke-like opacity. Previous studies have suggested that paraptosis is involved in the onset of early ARCC vacuolar degeneration. In this experiment, hydrogen peroxide (H<sub>2</sub>O<sub>2</sub>)-induced SRA01/04 cells were used to establish a paraptosis-like cell model, and the function and underlying mechanism of APEX1 in this cell model were explored. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) and Western Blot analyses were conducted to assess the expression of pertinent genes in SRA01/04. Confocal fluorescence microscopy, using ER-tracker kits, was applied to clarify the relationship between the endoplasmic reticulum and intracellular vacuoles. The co-IP assay was used to verify the interaction between APEX1 and P53. The GTRD database was employed to predict the putative target genes combined with P53, and CUT&RUN assay was employed to confirm the enrichment of the P53 and ATF6 promoters following APEX1 overexpression. Firstly, the pathological sections of the vacuolar degeneration zone in the lens cortex of ARCC patients exhibited fiber disarray and vacuole development. Meanwhile, the protein expression of Alix, a specific paraptosis inhibitor, was decreased in low-concentration H<sub>2</sub>O<sub>2</sub>-treated SRA01/04 cells. Secondly, we discovered that 4-PBA suppressed the expression of ATF6 and PERK. Moreover, overexpression of APEX1 in SRA01/04 cells improved endoplasmic reticulum morphology, inhibited the interaction between P53 and ATF6, and attenuated paraptosis in SRA01/04. APEX1 regulated P53 and then mediated ATF6 to affect the endoplasmic reticulum stress and paraptosis in H<sub>2</sub>O<sub>2</sub>-induced SRA01/04 cells.</div></div>","PeriodicalId":12177,"journal":{"name":"Experimental eye research","volume":"255 ","pages":"Article 110393"},"PeriodicalIF":3.0,"publicationDate":"2025-04-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143859823","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Limbal explant cultures on amniotic membrane: The effects of passaging the explants on cell phenotype

IF 3 2区医学 Q1 OPHTHALMOLOGY

Experimental eye research

Pub Date : 2025-04-16 DOI: 10.1016/j.exer.2025.110392

Mehmet Gurdal , Kemal Baysal , Ismet Durak , Ozlem Barut Selver

In vitro expansion of limbal epithelial stem cells (LESCs) while maintaining their characteristics has the potential to address the urgent need in ophthalmology clinics for the treatment of limbal stem cell deficiency (LSCD). Herein, we investigated the impact of explant passaging on the phenotype of LESCs cultured on human amniotic membrane (hAM). Following initial coverage of the hAM surface by cells (passage 0), the rabbit limbal explants underwent two additional passages. Expanded cells were then counted using a hemocytometer and examined by immunocytochemistry and RT-qPCR to assess markers associated with LESCs (ABCG2, P63, CK14, CXCR4, BMI-1, and vimentin) and differentiated LESCs (CK3 and connexin 43). The cell yield of passage 1 was the highest among all passages. Immunocytochemistry analysis revealed that the number of CK14-positive cells was similar across all passages; vimentin-positive cells were the lowest in passage 0, while vimentin-positive cells were the highest in passage 1; and CK3-positive cells were the highest in passage 0. RT-qPCR analysis revealed that CK3 and connexin 43 expression was significantly higher in passage 0 cells than in passage 2 cells; and CXCR4 and BMI-1 expressions were significantly higher in passage 1 cells than in passage 0 cells. Our data highlight that the passaging of limbal explant on hAM results in varying cell characteristics. The decrease in CK3 and increase in ABCG2 expression in cells obtained by passaging the limbal explant suggest that passaging could potentially enhance the stem cell population within the in vitro limbal explant culture on hAM.

{"title":"Limbal explant cultures on amniotic membrane: The effects of passaging the explants on cell phenotype","authors":"Mehmet Gurdal , Kemal Baysal , Ismet Durak , Ozlem Barut Selver","doi":"10.1016/j.exer.2025.110392","DOIUrl":"10.1016/j.exer.2025.110392","url":null,"abstract":"<div><div><em>In vitro</em> expansion of limbal epithelial stem cells (LESCs) while maintaining their characteristics has the potential to address the urgent need in ophthalmology clinics for the treatment of limbal stem cell deficiency (LSCD). Herein, we investigated the impact of explant passaging on the phenotype of LESCs cultured on human amniotic membrane (hAM). Following initial coverage of the hAM surface by cells (passage 0), the rabbit limbal explants underwent two additional passages. Expanded cells were then counted using a hemocytometer and examined by immunocytochemistry and RT-qPCR to assess markers associated with LESCs (ABCG2, P63, CK14, CXCR4, BMI-1, and vimentin) and differentiated LESCs (CK3 and connexin 43). The cell yield of passage 1 was the highest among all passages. Immunocytochemistry analysis revealed that the number of CK14-positive cells was similar across all passages; vimentin-positive cells were the lowest in passage 0, while vimentin-positive cells were the highest in passage 1; and CK3-positive cells were the highest in passage 0. RT-qPCR analysis revealed that CK3 and connexin 43 expression was significantly higher in passage 0 cells than in passage 2 cells; and CXCR4 and BMI-1 expressions were significantly higher in passage 1 cells than in passage 0 cells. Our data highlight that the passaging of limbal explant on hAM results in varying cell characteristics. The decrease in CK3 and increase in ABCG2 expression in cells obtained by passaging the limbal explant suggest that passaging could potentially enhance the stem cell population within the <em>in vitro</em> limbal explant culture on hAM.</div></div>","PeriodicalId":12177,"journal":{"name":"Experimental eye research","volume":"255 ","pages":"Article 110392"},"PeriodicalIF":3.0,"publicationDate":"2025-04-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143843749","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Identification of herpes simplex virus type 1 infection by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry

IF 3 2区医学 Q1 OPHTHALMOLOGY

Experimental eye research

Pub Date : 2025-04-15 DOI: 10.1016/j.exer.2025.110391

Chao Cheng , Yan Zong , Fang Duan , Ziyan Chen , Xiuping Liu , Kaili Wu

This study aimed to investigate whether Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) could identify Herpes simplex virus type 1 (HSV1) infection in samples in vitro and in vivo. MS spectra of supernatants and suspensions from infected human cornea epithelial (HCE) cell culture samples and infected samples of BALB/c mouse corneas were obtained by a VITEK® mass spectrometer. The discriminating peaks between infected and non-infected samples were used to establish discriminating superspectra (DSPc for cells and DSPm for corneas) by SARAMIS™ software. Another infected cells with two viral titers and infected cornea samples were used for blind testing against two DSPs. The results showed that automatic matching by the SARAMIS system revealed 28 discriminating peaks in HSV1-infected cells and 17 discriminating peaks in HSV1 keratitis, generating two discriminating superspectra (DSPs). Blind testing of virus-infected samples demonstrated a high positive identification rate for both in vitro and in vivo DSPs. The positive identification rate varied with viral titers, with cell suspensions exhibiting significantly higher rates compared to supernatants. Cluster analysis based on MS spectra revealed that there were more obvious differences between in vivo and in vitro samples compared to the differences between infected and non-infected samples. These findings suggest that MALDI-TOF MS can directly identify HSV1 in vitro or in vivo infected specimens, with higher positivity rates achieved when using cellular suspensions directly. This is an attempt on the method of virus detection, which shows potential for using MS to detect HSV1 infection or other virus infection in humans.

{"title":"Identification of herpes simplex virus type 1 infection by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry","authors":"Chao Cheng , Yan Zong , Fang Duan , Ziyan Chen , Xiuping Liu , Kaili Wu","doi":"10.1016/j.exer.2025.110391","DOIUrl":"10.1016/j.exer.2025.110391","url":null,"abstract":"<div><div>This study aimed to investigate whether Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) could identify Herpes simplex virus type 1 (HSV1) infection in samples <em>in vitro</em> and <em>in vivo</em>. MS spectra of supernatants and suspensions from infected human cornea epithelial (HCE) cell culture samples and infected samples of BALB/c mouse corneas were obtained by a VITEK® mass spectrometer. The discriminating peaks between infected and non-infected samples were used to establish discriminating superspectra (DSPc for cells and DSPm for corneas) by SARAMIS™ software. Another infected cells with two viral titers and infected cornea samples were used for blind testing against two DSPs. The results showed that automatic matching by the SARAMIS system revealed 28 discriminating peaks in HSV1-infected cells and 17 discriminating peaks in HSV1 keratitis, generating two discriminating superspectra (DSPs). Blind testing of virus-infected samples demonstrated a high positive identification rate for both <em>in vitro</em> and <em>in vivo</em> DSPs. The positive identification rate varied with viral titers, with cell suspensions exhibiting significantly higher rates compared to supernatants. Cluster analysis based on MS spectra revealed that there were more obvious differences between <em>in vivo</em> and <em>in vitro</em> samples compared to the differences between infected and non-infected samples. These findings suggest that MALDI-TOF MS can directly identify HSV1 <em>in vitro</em> or <em>in vivo</em> infected specimens, with higher positivity rates achieved when using cellular suspensions directly. This is an attempt on the method of virus detection, which shows potential for using MS to detect HSV1 infection or other virus infection in humans.</div></div>","PeriodicalId":12177,"journal":{"name":"Experimental eye research","volume":"255 ","pages":"Article 110391"},"PeriodicalIF":3.0,"publicationDate":"2025-04-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143850357","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Topical application of Cap-loaded hydrogels inhibits corneal neovascularization

IF 3 2区医学 Q1 OPHTHALMOLOGY

Experimental eye research

Pub Date : 2025-04-15 DOI: 10.1016/j.exer.2025.110390

Xiaoyang Xue , Xiaochuan Duan , Mengyuan Qin , Shoukuan Liu , Lin Su , Xin Cao , Hongquan Duan , Boshi Liu , Tianwen Ni , Xiaorong Li

Corneal neovascularization (CNV) is a significant risk factor for visual impairment. The efficiency and side effects of current CNV treatments, such as steroids and antivascular endothelial growth factor agents, are still debated. In addition, the bioavailability of topical drugs is usually hindered by tears, blinking, and the corneal anatomy. Therefore, finding a new therapeutic strategy is important. This study aimed to examine the function of the new therapeutic agent capmatinib (Cap), a highly selective inhibitor of MET that plays an important role in angiogenesis, in treating CNV. In this study, we first investigated the role of the HGF/c-MET axis in CNV and the therapeutic effect of Cap in a corneal alkali burn model. We synthesised a genipin-crosslinked gelatine-based hydrogel containing Cap (Cap-Gel). We observed a more significant therapeutic effect with the Cap-Gel than with Cap alone, as well as the alleviation of inflammatory infiltration and fibrosis. On day 14, the Cap-Gel group showed the most significant inhibition of corneal neovascularization, with the shortest neovessel length (0.48 ± 0.13 mm), smallest CNV area (3.77 ± 0.78 mm²), and lowest clinical assessment score (3.33 ± 0.52). Taken together, our results suggest that Cap-Gel could be a promising drug candidate for treating CNV.

{"title":"Topical application of Cap-loaded hydrogels inhibits corneal neovascularization","authors":"Xiaoyang Xue , Xiaochuan Duan , Mengyuan Qin , Shoukuan Liu , Lin Su , Xin Cao , Hongquan Duan , Boshi Liu , Tianwen Ni , Xiaorong Li","doi":"10.1016/j.exer.2025.110390","DOIUrl":"10.1016/j.exer.2025.110390","url":null,"abstract":"<div><div>Corneal neovascularization (CNV) is a significant risk factor for visual impairment. The efficiency and side effects of current CNV treatments, such as steroids and antivascular endothelial growth factor agents, are still debated. In addition, the bioavailability of topical drugs is usually hindered by tears, blinking, and the corneal anatomy. Therefore, finding a new therapeutic strategy is important. This study aimed to examine the function of the new therapeutic agent capmatinib (Cap), a highly selective inhibitor of MET that plays an important role in angiogenesis, in treating CNV. In this study, we first investigated the role of the HGF/c-MET axis in CNV and the therapeutic effect of Cap in a corneal alkali burn model. We synthesised a genipin-crosslinked gelatine-based hydrogel containing Cap (Cap-Gel). We observed a more significant therapeutic effect with the Cap-Gel than with Cap alone, as well as the alleviation of inflammatory infiltration and fibrosis. On day 14, the Cap-Gel group showed the most significant inhibition of corneal neovascularization, with the shortest neovessel length (0.48 ± 0.13 mm), smallest CNV area (3.77 ± 0.78 mm<sup>2</sup>), and lowest clinical assessment score (3.33 ± 0.52). Taken together, our results suggest that Cap-Gel could be a promising drug candidate for treating CNV.</div></div>","PeriodicalId":12177,"journal":{"name":"Experimental eye research","volume":"255 ","pages":"Article 110390"},"PeriodicalIF":3.0,"publicationDate":"2025-04-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143859822","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

TGFβ2 alters segmental outflow and ECM ultrastructure in the trabecular meshwork

IF 3 2区医学 Q1 OPHTHALMOLOGY

Experimental eye research

Pub Date : 2025-04-10 DOI: 10.1016/j.exer.2025.110377

Timur Mavlyutov, Samer E. Bilal, Justin J. Myrah, Kelsey M. Mathers, Taylor Y. Lee, Colleen M. McDowell

TGFβ2 is a well-known contributor to extracellular matrix (ECM) changes in the trabecular meshwork (TM). TGFβ2 is increased in the aqueous humor (AH) of primary open angle glaucoma patients and the addition of TGFβ2 to primary human TM cells in culture induces pathogenic changes similar to what is seen in the TM of ocular hypertensive and glaucomatous eyes. Overexpression of a bioactivated form of TGFβ2 using adenovirus 5 (Ad5.TGFβ2) in the TM has previously been described as an inducible mouse model of ocular hypertension and has been utilized for multiple studies to help understand the pathogenies of TGFβ2-induced TM damage and elevated intraocular pressure (IOP). Ad5.TGFβ2 is known to elevate IOP, decrease outflow facility, and increase expression of ECM proteins in the TM. Here, we further analyze the effects of overexpression of TGFβ2 by Ad5 in the TM. We found Ad5.TGFβ2 increases expression of macrophage marker Iba1 and increases expression of ECM proteins fibronectin and collagen 1 compared to Ad5.Null injected controls. In addition, overexpression of TGFβ2 by Ad5 led to a decrease in segmental AH flow regions compared to Ad5.Null control eyes. Ultrastructure analysis of the Ad5.TGFβ2 injected eyes also show significantly more areas occupied by ECM material as well as the development of more smaller giant vacuoles compared to Ad5.Null injected eyes. These data in combination with prior research using Ad5.TGFβ2, establish the use of intraocular injection of Ad5.TGFβ2 as an appropriate mouse model of ocular hypertension to study aqueous humor outflow and its mechanisms.

{"title":"TGFβ2 alters segmental outflow and ECM ultrastructure in the trabecular meshwork","authors":"Timur Mavlyutov, Samer E. Bilal, Justin J. Myrah, Kelsey M. Mathers, Taylor Y. Lee, Colleen M. McDowell","doi":"10.1016/j.exer.2025.110377","DOIUrl":"10.1016/j.exer.2025.110377","url":null,"abstract":"<div><div>TGFβ2 is a well-known contributor to extracellular matrix (ECM) changes in the trabecular meshwork (TM). TGFβ2 is increased in the aqueous humor (AH) of primary open angle glaucoma patients and the addition of TGFβ2 to primary human TM cells in culture induces pathogenic changes similar to what is seen in the TM of ocular hypertensive and glaucomatous eyes. Overexpression of a bioactivated form of TGFβ2 using adenovirus 5 (Ad5.TGFβ2) in the TM has previously been described as an inducible mouse model of ocular hypertension and has been utilized for multiple studies to help understand the pathogenies of TGFβ2-induced TM damage and elevated intraocular pressure (IOP). Ad5.TGFβ2 is known to elevate IOP, decrease outflow facility, and increase expression of ECM proteins in the TM. Here, we further analyze the effects of overexpression of TGFβ2 by Ad5 in the TM. We found Ad5.TGFβ2 increases expression of macrophage marker Iba1 and increases expression of ECM proteins fibronectin and collagen 1 compared to Ad5.Null injected controls. In addition, overexpression of TGFβ2 by Ad5 led to a decrease in segmental AH flow regions compared to Ad5.Null control eyes. Ultrastructure analysis of the Ad5.TGFβ2 injected eyes also show significantly more areas occupied by ECM material as well as the development of more smaller giant vacuoles compared to Ad5.Null injected eyes. These data in combination with prior research using Ad5.TGFβ2, establish the use of intraocular injection of Ad5.TGFβ2 as an appropriate mouse model of ocular hypertension to study aqueous humor outflow and its mechanisms.</div></div>","PeriodicalId":12177,"journal":{"name":"Experimental eye research","volume":"255 ","pages":"Article 110377"},"PeriodicalIF":3.0,"publicationDate":"2025-04-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143824195","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Effects of adenosine triphosphate and coenzyme Q10 on potential hydroxychloroquine-induced retinal damage in rats.

IF 3 2区医学 Q1 OPHTHALMOLOGY

Experimental eye research

Pub Date : 2025-04-09 DOI: 10.1016/j.exer.2025.110387

Ahmet Duhan Ozbay , Ahmet Mehmet Somuncu , Ibrahim Cicek , Bulent Yavuzer , Seval Bulut , Gulbaniz Huseynova , Tugba Bal Tastan , Mine Gulaboglu , Halis Suleyman

This study aimed to investigate biochemically and histopathologically the protective effect of adenosine triphosphate (ATP) and coenzyme Q10 (CoQ10) against potential hydroxychloroquine (HCQ)-induced retinal damage in rats. Twenty-four male albino Wistar-type rats were randomly separated into four groups: healthy (HG), receiving HCQ (HQG), receiving ATP + HCQ (AHQ), and receiving CoQ10 + HCQ (CoQHQ). ATP (4 mg/kg, intraperitoneal) was given to the AHQ, and CoQ10 (10 mg/kg, oral) to the CoQHQ. Rats in the HQG, AHQ, and CoQHQ were given HCQ (120 mg/kg, oral) 1 h after administering ATP and CoQ10. Treatments continued once a day for seven days. On the 8th day, the rats were sacrificed with 50 mg/kg sodium thiopental, and the eyes were removed. Malondialdehyde (MDA), total glutathione (tGSH), superoxide dismutase (SOD), and catalase (CAT), and interleukin-6 (IL-6) levels were measured in the retrieved eye tissues and retinal tissues were assessed histopathologically. An increase in MDA and IL-6 levels and a decrease in tGSH, SOD, and CAT levels were detected in the eye tissues of the HQGcompared to the HG. HCQ-induced changes in oxidant and antioxidant levels were significantly suppressed by ATP and CoQ10 treatment. ATP was more successful than CoQ10 in this inhibition. Severe damage was observed in the eye tissues of the HQG group, whereas the damage was mild in the AHQ and moderate in the CoQHQ. Although both ATP and CoQ10 have the potential to be effective in the prevention of HCQ-induced retinal damage through antioxidative activity, ATP appears to be the more preferable treatment approach.

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引用次数: 0

Oxygen uptake at the ocular surface in diabetic animals is impaired in response to central corneal injury

IF 3 2区医学 Q1 OPHTHALMOLOGY

Experimental eye research

Pub Date : 2025-04-09 DOI: 10.1016/j.exer.2025.110384

Ana M. Sandoval-Castellanos , Sun Qin , Li Ma , Fernando Ferreira , Brian Reid , Min Zhao

Poor wound healing is one of the most devastating complications in late-stage diabetic patients. The avascular cornea does not rely on circulation for its oxygen consumption, uptaking it mainly from the atmosphere. Previous studies demonstrated that oxygen uptake (O₂U) in diabetic experimental animals and patients is significantly lower than in the non-diabetic condition. Our recent experiments show that upon wounding of the central cornea the O₂U decreased across the ocular surface, followed by two increases at 6–24 h, and at 72 h, which appeared to be important for proper wound healing. It is however not known whether the two distinct O₂U increases are maintained in diabetic ocular surface in response to corneal injury. In this study, we used an optic-fiber oxygen micro-sensor to measure O₂U across the ocular surface of streptozotocin (STZ)- induced diabetic mice and age-matched control mice following injury to the central cornea. We found that the injury causes an immediate and substantial reduction of O₂U across the ocular surface. O₂U in non-diabetic corneas increases at 2–6 h post wounding (hpw), decreasing again before the second rise to peak at 72 hpw, especially at the limbus. O₂U in the diabetic cornea decreases more markedly than that of non-diabetic control. This defective diabetic O₂U persisted, precluding the two dynamic rises in O₂U, leading to a failure in recovery. Altogether, our results suggest a previously unknown mechanism of a defective O₂U response to injury in the diabetic ocular surface, which warrants further research and may lead to new therapeutic paths.

伤口愈合不良是糖尿病晚期患者最严重的并发症之一。无血管角膜的耗氧量不依赖血液循环，主要从大气中摄取。以前的研究表明，糖尿病实验动物和患者的摄氧量（O2U）明显低于非糖尿病状态。我们最近的实验表明，中央角膜受伤后，整个眼表的 O2U 会下降，随后在 6-24 小时和 72 小时内会有两次上升，这似乎对伤口的正常愈合很重要。然而，我们还不知道糖尿病患者的眼表在角膜损伤后是否会维持这两种不同的 O2U 增加。在这项研究中，我们使用光导纤维氧微量传感器测量了链脲佐菌素（STZ）诱导的糖尿病小鼠和年龄匹配的对照组小鼠在角膜中央损伤后眼表的 O2U。我们发现，损伤会导致整个眼表面的 O2U 立即大幅下降。非糖尿病角膜中的 O2U 在受伤后 2-6 小时内上升，在第二次上升之前再次下降，在 72 小时内达到峰值，尤其是在角膜缘。与非糖尿病对照组相比，糖尿病角膜的 O2U 下降更为明显。这种糖尿病 O2U 缺陷持续存在，阻碍了 O2U 的两次动态上升，导致恢复失败。总之，我们的研究结果表明，糖尿病眼表对损伤的 O2U 反应缺陷是一种以前未知的机制，值得进一步研究，并可能带来新的治疗途径。

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引用次数: 0

Melatonin ameliorates retinal neurovascular degeneration in Rd1 mice by inhibiting oxidativestress 褪黑激素通过抑制氧化应激改善 Rd1 小鼠视网膜神经血管变性的情况

IF 3 2区医学 Q1 OPHTHALMOLOGY

Experimental eye research

Pub Date : 2025-04-09 DOI: 10.1016/j.exer.2025.110388

Aoxiang Wang, Haichun Li, Yue Wu, Tong Wang, Ping Lian

Oxidative stress has been involved in the occurrence of retinal photoreceptor degeneration and retinal vascular dysfunctions. This study investigated the effects of melatonin (MLT) on neurovascular changes in rd1 mice, evaluating its therapeutic potential as an antioxidant for retinal degeneration. MLT was administered to rd1 mice at postnatal day 7 (P7), and retinal vascular alterations were assessed using retina flatmounts, while neural and functional changes were evaluated through frozen sections and electroretinography at P14. In vitro, human retinal microvascular endothelial cells (HRMECs) were treated with MLT to counteract oxidative stress induced by H2O2. Analyses included assessments of cell function, apoptosis, oxidative stress, and inflammatory markers in both in vivo and in vitro models. The results demonstrated that MLT significantly improved retinal vascular densities in the deep and superficial layers at P14 and P21, though not fully restoring them to wild-type levels. Additionally, MLT exerted protective effects against photoreceptor degeneration, oxidative stress, and inflammation, partially preserving retinal function. In vitro, MLT alleviated functional abnormalities and reduced cell death in HRMECs by decreasing reactive oxygen species levels. These findings suggest that MLT holds promise as a therapeutic approach for retinal degeneration by mitigating oxidative stress, thereby protecting photoreceptors and retinal vasculature. This underscores the importance of vascular preservation in developing therapeutic strategies for retinal degenerative diseases.

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引用次数: 0